本論文は、異種生体分子を酵素反応を用いて部位特異的にアセンブルする技術の基礎研究から応用に至る展開を発表したものである。意図する機能を発現するための分子設計指針から、特異な構造を有する生体分子の調製、得られたハイブリッド分子による高感度遺伝子検出試薬の実用化（JST育成研究成果）とセルロース系バイオマス分解触媒としての利用価値（JST-ALCA研究成果）について言及した。

Language：English   Publishing type：Research paper (scientific journal)  

DOI： https://doi.org/10.1021/acsinfecdis.2c00052

Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.jbiosc.2021.12.004

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DOI： 10.1021/acs.bioconjchem.1c00235

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DOI： 10.1016/j.pep.2020.105730

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DOI： 10.1016/j.ejpb.2018.01.020

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DOI： 10.1016/j.febslet.2005.02.064

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DOI： 10.1021/bm050193o

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DOI： 10.1021/bm0494895

Language：English   Publisher：Chemical Communications  

The fusion protein of an engineered zymogen of microbial transglutaminase (EzMTG) with a protein G variant, EzMTG-pG, enabled the proximity-based, tag-free labeling of Lys65 in the heavy chain of a native IgG antibody (trastuzumab) with a Gln-donor peptidyl substrate functionalized with a fluorescent molecule.

DOI： 10.1039/d4cc01728e

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Publisher：Waste and Biomass Valorization  

Purpose: This work evaluated the profiles of Industrial agar seaweed waste biomass (ISWB) and pretreated ISWB by using deep eutectic solvent (DESs) and NaOH. The profiles of nanocellulose obtained from pretreated ISWB (with various types of hydrogen bond donors of choline chloride (ChCl)-based hydrated DES) were also evaluated. Methods: ISWB underwent pre-treatment with either 100% wt. DES (1:1 molar ratio of ethylene glycol: citric acid) or 10% wt. NaOH at 80 °C to 90 °C. Subsequent treatment involved 10% wt. DES or ChCl-based hydrated DES with various HBDs (oxalic acid, citric acid, and urea) in a 1:2 molar ratio to produce nanocellulose. Results: Cellulose nanofibers isolated by the CE-DES approach exhibited a substantially higher yield compared with those obtained via the CE-NaOH method. CE-DES method induced swelling of ISWB and facilitate production of cellulose nanofibers. SEM images of samples indicates the presence of fibrous nanostructures across the majority of the specimens. Fourier-transform infrared (FTIR) spectroscopy demonstrated uniformity among all samples; they exhibited a consistent fingerprint pattern. X-ray diffraction (XRD) spectra indicated the presence of cellulose Type I in cellulose nanofibers produced by CE–DES, but not definitively in nanocellulose produced by the CE–NaOH method. Conclusion: Cellulose nanofibers can be isolated from ISWB by using various DES and exhibits substantial characteristics of cellulose nanofibers from different ISWB pretreatment protocols. Graphical Abstract: (Figure presented.)

DOI： 10.1007/s12649-024-02499-z

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Language：English   Publisher：Molecules  

Herein, we report a transdermal patch prepared using an ionic liquid-based solid in oil (IL-S/O) nanodispersion and a pressure-sensitive adhesive (PSA) to deliver the macromolecular antigenic protein, ovalbumin (OVA). The IL-S/O nanodispersion and a PSA were first mixed at an equal weight ratio, then coated onto a release liner, and covered with a support film. To evaluate the effect of the PSA, three types of PSAs, DURO-TAK 87-4098, DURO-TAK 87-4287, and DURO-TAK 87-235A, were used to obtain the corresponding IL-S/O patches SP-4098, SP-4287, and SP-235A, respectively. The prepared IL-S/O patches were characterized for surface morphology, viscoelasticity, and moisture content. In vitro skin penetration and in vivo immunization studies of the IL-S/O patches were performed using Yucatan micropig skin and the C57BL/6NJc1 mice model, respectively. The SP-4098 and SP-4287 delivered 5.49-fold and 5.47-fold higher amounts of drug compared with the aqueous formulation. Although both patches delivered a similar amount of drug, SP-4287 was not detached fully from the release liner after 30 days, indicating low stability. Mice immunized with the OVA-containing SP-4098 produced a 10-fold increase in anti-OVA IgG compared with those treated with an aqueous formulation. These findings suggested that the IL-S/O patch may be a good platform for the transdermal delivery of antigen molecules.

DOI： 10.3390/molecules29132995

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DOI： 10.1016/j.jbiosc.2024.04.001

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Language：English   Publisher：(公社)日本生物工学会  

Publisher：Fisheries Science  

Ulva is an unutilized green seaweed that grows in the intertidal zone around the coast of Java Island, Indonesia. In this study, U. lactuca samples collected from three regions in south of Java Island (Cihara, Surade, and Tepus) were studied in terms of their chemical composition, physical properties and its bioactivity, to determine the best regions for establishing seaweed industries. The chemical characteristics differed significantly among different regions, where the seaweed from Tepus, Surade and Cihara had the highest content of protein (22.93%), carbohydrate (61.58%), and mineral (28.72%), respectively. The amino acids were dominated with L-aspartic acid and L-glutamic acid. All U. lactuca samples contained abundant pigments, such as chlorophylls and carotenoids, especially samples from Tepus. The highest content of crude ulvan was found in Surade seaweed (26.9%). Chemical and physical analyses showed the presence of S = O and C–O–S functional groups in ulvan, a sulfated polysaccharide unique to Ulva sp., with thermal degradation up to 220 °C. Crude ulvan from Surade and Tepus seaweed exhibited bioactivity to support proliferation of fibroblast cells at 100 and 1000 ppm, respectively. Based on the properties of U. lactuca, Tepus and Surade were identified as potential sites to establish aquaculture and/or processing industries.

DOI： 10.1007/s12562-024-01799-6

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Publisher：ASEAN Journal of Chemical Engineering  

Ulva lactuca is a green seaweed commonly called sea lettuce and contains sulphated polysaccharides that have biological activity. Despite its lack of ecological sustainability, strong acids (HCl) and strong bases (NaOH) are widely used as solvents in the conventional extraction process for ulvan. Deep eutectic solvent (DES) is an alternative to ulvan extraction solvent, which is more environmentally friendly and has low toxicity. This study aims to assess the efficacy of peracetic acid (PAA) pretreatment and post-treatment in enhancing the quality of extracted ulvan using a DES-based solvent in the extraction process. Ulvan extraction using DES with a temperature of 85-95°C for 1 hour and adding 2% PAA pretreatment, 0.1%, 0.5%, and 2% PAA post-treatment were conducted. PAA pre-and post-treatment was carried out with a 1:10 (w/v) ratio at room temperature for 30 minutes. The analysis included moisture content, yield, functional groups, sulphate content, and color. The test results revealed that ulvan treated with 2% PAA pretreatment had the highest moisture content and sulphate content, at 18.71% and 33.39%, respectively, while ulvan treated with 0.1% PAA post-treatment had the highest yield, at 41.96%. Adding peracetic acid concentration can increase the color quality of the ulvan. PAA pre-and post-treatment had a significant effect on all ulvan quality parameters.

DOI： 10.22146/ajche.12272

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Herein, we report ethosome (ET) formulations composed of a safe amount of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC)-based ionic liquid with various concentrations of ethanol as a carrier for the transdermal delivery of a high molecular weight drug, insulin. The Insulin-loaded ET vesicles exhibited long-term stability compared to conventional DMPC ETs, showing significantly higher drug encapsulation efficiency and increased skin permeation ability.

DOI： 10.1039/d3cc06130b

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DOI： 10.1101/2024.03.04.583322

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Ionic liquid (IL)-based transdermal formulations have gained significant attention because of their high skin penetration efficiency and ease of design. However, many IL-based formulations exhibit low viscosity, which hinders direct administration and leads to less efficient drug delivery. Therefore, there is a pressing need for specialized formulations with improved viscosity and efficacy as a drug delivery platform. In this study, we report a ready-to-use transdermal patch system utilizing IL-in-oil (IL/O) microemulsions to deliver acyclovir, a sparingly soluble antiviral drug. To prepare the patch system, a mixture containing equal weights of IL/O microemulsions and adhesive (Durotak 87–4098) was homogenized, subsequently coated on a liner, and covered by a support film. These patches were produced with varying thicknesses (100 μm, 200 μm, 300 μm, and 500 μm) via a solvent evaporation technique. A 24-hour in vitro skin permeation study revealed that the thinnest patch (100 μm) delivered more than 45 % of the loaded drug. Drug delivery rate gradually reduced as the patch thickness increased, accounting for 22 % and 14 % drug permeation by 200 μm and 300 μm patches respectively. A 30-day stability study at room temperature demonstrated that neither the physical appearance nor the active ingredient content changed, validating the physical and chemical stability of the patches. Skin permeation mechanism by Fourier transform infrared spectroscopic analysis of the skin suggested that the IL/O patch possesses the capability to modify the lipid barrier in the stratum corneum thus facilitating drug diffusion. In vitro skin irritation studies on reconstructed human epidermal tissue and Yucatan micropig skin revealed that the patch exhibits no skin irritation. We believe that the adhesive nature of this newly developed IL-based patch holds significant promise as a transdermal drug delivery platform.

DOI： 10.1016/j.molliq.2024.124184

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Publishing type：Research paper (scientific journal)   Publisher：MYU K.K.  

DOI： 10.18494/SAM4816

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Language：English   Publisher：(公社)日本生物工学会  

ペルオキシダーゼ触媒によるクロスリンク形成を介して生理的な条件下でゲル化を受けるアルブミン派生物を合成した。アルブミンをカルボジイミドを用いてフェノール性水酸基(Alb-Ph-OH)で修飾し、置換度がゲル化に及ぼす影響を解析した。Alb-Ph-OHヒドロゲルの多様な特性、すなわち、ゲル化時間、膨張率、孔径、貯蔵弾性率、および酵素的な分解性は置換度とポリマー濃度の調整により容易に調節可能であった。Alb-Ph-OHヒドロゲル中にカプセル化された細胞の生存率は高かった。

DOI： 10.1101/2023.10.09.561484

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DOI： 10.1016/j.jbiosc.2023.09.007

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DOI： https://doi.org/10.1101/2023.10.09.561484

Repository Public URL： https://hdl.handle.net/2324/7236919

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DOI： 10.1021/acsami.3c03900

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Language：Others   Publishing type：Research paper (scientific journal)   Publisher：The Chemical Society of Japan  

DOI： 10.1246/cl.230156

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DOI： 10.1016/j.jbiosc.2023.03.015

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Language：Japanese   Publisher：The Society for Biotechnology, Japan  

DOI： 10.34565/seibutsukogaku.101.3_117

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Malaria is a mosquito-borne infectious disease that is widespread in developing countries. Malaria vaccines are important in efforts to eradicate malaria; however, vaccines are usually administered by injection, which requires medical personnel and has a risk of causing infection. Transdermal vaccines can be administered without damaging the skin and thus are ideal for the prevention of malaria. However, the stratum corneum forms a "brick and mortar" like structure in which stratum corneum cells are embedded in a hydrophobic matrix composed of lipids, which strongly inhibits the permeation of hydrophilic substances. In the present study, we designed a transdermal vaccine against vivax malaria using a solid-in-oil (S/O) dispersion. The S/O dispersion of a transmission blocking vaccine candidate, Pvs25 from Plasmodium vivax, showed higher skin penetration than that of the aqueous solution. Mice immunized with the S/O dispersion generated antibodies at similar titers as the mice immunized by injection, over the mid- to long-term. These results provide information for the development of transdermally administered malaria vaccines toward the eradication of malaria.

DOI： 10.1016/j.xphs.2022.10.031

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This study investigated the localization of artificial peptide supramolecular fibers in liquid-liquid phase separation (LLPS). Hierarchical organization led to the localization of supramolecules in LLPS droplets. Moreover, proteins were recruited into confined droplets by the physical adsorption of proteins on the supramolecules, enabling an enhanced cascade reaction.

DOI： 10.1039/d2cc05910j

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Cytoplasm contains high concentrations of biomacromolecules. Protein behavior under such crowded conditions is reportedly different from that in an aqueous buffer solution, mainly owing to the effect of volume exclusion caused by the presence of macromolecules. Using a crosslinking reaction catalyzed by microbial transglutaminase (MTG) as a model, we herein systematically determined how the substrate size affects enzymatic activity in both dilute and crowded solutions of dextran. We first observed a threefold reduction in MTG-mediated crosslinking of a pair of small peptide substrates in 15 wt% dextran solution. In contrast, when proteinaceous substrates were involved, the crosslinking rates in 15 wt% dextran solutions accelerated markedly to levels comparable with the level in the absence of dextran. Our results provide new insights into the action of enzymes with regard to macromolecular substrates under crowded conditions, of which the potential utility was demonstrated by the formation of highly crosslinked protein polymers.

DOI： 10.1039/d2ob01549h

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Myocardial damage caused by the newly emerged coronavirus (SARS-CoV-2) infection is one of the key determinants of COVID-19 severity and mortality. SARS-CoV-2 entry to host cells is initiated by binding with its receptor, angiotensin-converting enzyme (ACE) 2, and the ACE2 abundance is thought to reflect the susceptibility to infection. Here, we report that ibudilast, which we previously identified as a potent inhibitor of protein complex between transient receptor potential canonical (TRPC) 3 and NADPH oxidase (Nox) 2, attenuates the SARS-CoV-2 spike glycoprotein pseudovirus-evoked contractile and metabolic dysfunctions of neonatal rat cardiomyocytes (NRCMs). Epidemiologically reported risk factors of severe COVID-19, including cigarette sidestream smoke (CSS) and anti-cancer drug treatment, commonly upregulate ACE2 expression level, and these were suppressed by inhibiting TRPC3-Nox2 complex formation. Exposure of NRCMs to SARS-CoV-2 pseudovirus, as well as CSS and doxorubicin (Dox), induces ATP release through pannexin-1 hemi-channels, and this ATP release potentiates pseudovirus entry to NRCMs and human iPS cell-derived cardiomyocytes (hiPS-CMs). As the pseudovirus entry followed by production of reactive oxygen species was attenuated by inhibiting TRPC3-Nox2 complex in hiPS-CMs, we suggest that TRPC3-Nox2 complex formation triggered by panexin1-mediated ATP release participates in exacerbation of myocardial damage by amplifying ACE2-dependent SARS-CoV-2 entry.

DOI： 10.3390/ijms24010102

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Publisher：Journal of Molecular Liquids  

Luteolin (LUT), an important flavonoid, shows excellent antioxidant and antimicrobial properties. However, its low solubility and stability in water, as well as challenges related to its formulation, have limited its use in food preservation. Here, we develop a highly water-soluble LUT-loaded micellar formulation (LUT-MF) using a solid-in-water nanodispersion technique involving emulsification and lyophilization processes. LUT solubilized in an organic phase comprising acetone, methanol, and cyclohexane (5:10:85 v/v) was emulsified with an aqueous solution of a surface-active ionic liquid, choline oleate ([Cho][Ole]), and the oil in water emulsion was lyophilized to remove the organic phase and water and obtain a solid LUT-[Cho][Ole] complex. LUT-[Cho][Ole] complex was dispersed again in water to prepare the LUT-MF formulation with good stability. Dynamic light scattering and transmission electron microscopy confirmed that the LUT-MF formed spherical micelles with a mean particle size of approximately 73 nm. LUT-MF showed excellent encapsulation efficiency (94.3%) and enhanced aqueous solubility. An in vitro release study in PBS containing 30% ethanol demonstrated 3.94 times higher release of LUT from LUT-MF than from LUT suspension after 24 h. The developed formulation showed better antioxidant activity than both free LUT and ascorbic acid. LUT-MF demonstrated higher antibacterial activity than sodium benzoate. Food preservation studies revealed that treating strawberries with LUT-MF effectively preserved color and prevented shriveling and decay over 8 days storage at 10 °C, compared with sodium benzoate treatment. The results suggest that LUT-MF is a potential natural preservative alternative to conventional toxic chemical preservatives used in the food industry.

DOI： 10.1016/j.molliq.2022.120151

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Language：English   Publisher：(公社)日本生物工学会  

市販のアンフォテリシンB(AMB)製剤と表面電荷が異なるリポソームを組合せてハイブリッドリポソーム製剤を調製し、抗真菌活性の重要な要因を探索した。AMB含有リポソーム製剤(AMB-LF)の粒子サイズ分布とゼータ電位を含む特性評価では、陰イオン性と中性のAMB-LFはAMBを安定的にカプセル化できた。アニオン性または中性AMB-LFと未修飾LysMの組合せではAMBのMICが低下した。中性AMB-LFとPteris ryukyuensisキチナーゼ由来の人工パルミトイル化キチン結合ドメインの組合せではMICは更に減少し、中性AMB-LF単独と比較して最大15倍になった。以上より、脂質修飾性蛋白質結合剤と組合せたAMBの脂質基盤リポソーム製剤の真菌感染に対する有用であると考えられた。

Language：English   Publishing type：Research paper (scientific journal)  

Transdermal drug delivery systems (TDDSs) may be useful for preventing various diseases including cancer. However, the stratum corneum (SC) inhibits the permeation of foreign particles into the skin. To obtain an effective TDDS, we developed a protein-containing nanocarrier (PCNC) comprising an antigenic protein (ovalbumin/OVA) stabilized by a combination of surfactants, i.e., a lipid-based surface-active ionic liquid and Tween-80. The PCNC was lyophilized to remove water and cyclohexane and then dispersed in isopropyl myristate. It is biocompatible both in vitro and in vivo, and is suitable for use in a therapeutic TDDS. The skin permeability of the PCNC was significantly (p < 0.0001) enhanced, and the transdermal distribution and transdermal flux of the OVA delivery system were 25 and 28 times greater, respectively, than those of its aqueous formulation. The PCNC disrupted the order of lipid orientation in the skin's SC and increased intercellular protein delivery. It demonstrated effective antitumor activity, drastically (p < 0.001) suppressed tumor growth, increased mouse survival rates, and significantly (p < 0.001) stimulated the OVA-specific tumor immune response. The PCNC also increased the number of cytotoxic T cells expressing CD8 antibodies on their surfaces (CD8 + T-cells) in the tumor microenvironment. These findings suggest that PCNCs may be promising biocompatible carriers for transdermal antigenic protein delivery in tumor immunotherapy.

DOI： 10.1021/acsabm.2c00061

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Combinations of antifungal drugs can have synergistic antifungal activity, achieving high therapeutic efficacy while minimizing the side effects. Amphotericin B (AMB) has been used as a standard antifungal drug for fungal infections; however, because of its high toxicity, new strategies to minimize the required dose are desirable. Chitinases have recently received attention as alternative safe antifungal agents. Herein, we report the combination of palmitoylated chitinase domains with AMB to enhance the antifungal activity. The chitin-binding domain (LysM) from Pteris ryukyuensis chitinase was site-specifically palmitoylated by conjugation reaction catalyzed by microbial transglutaminase. The palmitoylated LysM (LysM-Pal) exhibited strong antifungal activity against Trichoderma viride, inhibiting the growth completely at a concentration of 2 μM. This antifungal effect of LysM-Pal was mainly due to the effect of anchoring of palmitic acid motif to the plasma membrane of fungi. A combination of AMB with LysM-Pal resulted in synergistic enhancement of the antifungal activity. Intriguingly, LysM-Pal exhibited higher level of antifungal activity enhancement than palmitoylated catalytic domain (CatD) and fusion of LysM and CatD. Addition of 0.5 μM LysM-Pal to AMB reduced the minimal inhibition concentration of AMB to 0.31 μM (2.5 μM without LysM-Pal). The possible mechanism of the synergistic effect of AMB and LysM-Pal is destabilization of the plasma membrane by anchoring of palmitic acid and ergosterol extraction by AMB and destabilization of the chitin layer by LysM binding. The combination of LysM-Pal with AMB can drastically reduce the dose of AMB and may be a useful strategy to treat fungal infections.

DOI： 10.1021/acsinfecdis.2c00052

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Publisher：Journal of Molecular Liquids  

Ionic liquids (ILs) have gained great attention as potent solubilizing agents of poorly soluble bioactive compounds. The use of Luteolin (LUT) – a natural bioactive compound – is limited because of its poor aqueous solubility. To address this issue, this study reports potential ILs for effectively solubilizing LUT at ambient temperature. First, promising ILs were screened from a potential 180 ILs that were proposed by the combination of 20 standard amino acid ethyl esters cations and nine phenolic acids anions using the conductor-like screening model for real solvents (COSMO-RS); the activity coefficients of LUT at infinite dilution in ILs were used as the screening criteria. Four proline ethyl ester phenolate ILs (PEEP-ILs) composed of a biocompatible proline ethyl ester and bioactive phenolic acids (trans-ferulic, vanillic, p-coumaric, and 4-hydroxybenzoic acid) were selected and synthesized. The synthesized PEEP-ILs were characterized through 1H NMR, FT-IR, elemental analysis, TGA, and DSC. The experimental solubility of LUT in PEEP-ILs (60% IL in water) showed excellent solubility. Among the PEEP-ILs, proline ethyl ester ferulate (IL[ProEt][Fer]) exhibited the best LUT solubility, followed by ILs with p-coumarate (IL[ProEt][Cou]), vanillate (IL[ProEt][Van]), and 4-hydroxybenzoate (IL[ProEt][Ben]). 1H NMR assessment and COSMO-RS prediction confirmed that the ILs facilitated the solubilization of LUT through multiple hydrogen bonding, π − π, and cation − π interactions between the LUT and the ILs. The biocompatibility study revealed that PEEP-ILs were relatively harmless and low toxicity compared with conventional imidazolium, pyridinium, pyrrolidinium, piperidinium, and morpholinium-based ILs. The LUT dissolved in the newly-synthesized PEEP-ILs and demonstrated better organoleptic properties on red apple slices. These results indicate that PEEP-ILs can be potential green alternatives to conventional toxic solvents for dissolving poorly water-soluble bioactive natural preservatives.

DOI： 10.1016/j.molliq.2021.118103

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.biomaterials.2022.121385

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Language：Japanese   Publishing type：Research paper (scientific journal)  

Transcutaneous drug delivery is a promising method in terms of drug repositioning and reformulation because of its non-invasive and easy-to-use features. To overcome the skin barrier, which is the biggest challenge in transcutaneous drug delivery, a number of techniques, such as microemulsion, solid-in-oil dispersions and liposomes, have been studied extensively. However, the low viscosity of these formulations limits drug retention on the skin and reduces patient acceptability. Although viscosity can be increased by adding a thickening reagent, such an addition often alters formulation nanostructures and drug solubility, and importantly, decreases skin permeability. In this study, a gel-like lyotropic liquid crystal (LLC) was used as a tool to enhance skin permeability. In particular, we prepared 1-monolinolein (ML)-based LLCs with different water contents. All LLCs significantly enhanced skin permeation of a peptide drug, an epitope peptide of melanoma, despite their high viscoelasticity. Fourier transform infra-red spectroscopic analysis of the skin surface treated with the LLCs revealed that the gyroid geometry more strongly interacted with the lamellar structure inside the stratum corneum (SC) than the diamond geometry. Finally, as the result of the in vivo tumor challenge experiment using B16F10 melanoma-bearing mice, the LLC with the gyroid geometry showed stronger vaccine effect against tumor than a subcutaneous injection. Collectively, ML-based LLCs, especially with the gyroid geometry, are a promising strategy to deliver biomacromolecules into skin. Statement of significance: Transcutaneous drug delivery is a promising method for drug repositioning and reformulation because of its non-invasive and easy-to-use features. To overcome the skin barrier, which is the biggest challenge in transcutaneous drug delivery, we used a gel-like lyotropic liquid crystal (LLC) as a novel tool to enhance skin permeability. In this paper, we demonstrated that an LLC with a specific liquid crystalline structure has the highest skin permeation enhancement effect for a peptide antigen as a model drug. Moreover, the peptide antigen-loaded LLC showed a vaccine effect that was comparable to a subcutaneous injection in vivo. This study provides a basis for designing a transcutaneous delivery system of peptide drugs with LLC.

DOI： 10.1016/j.actbio.2021.11.008

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Scopus

PubMed

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Royal Society of Chemistry (RSC)  

Self-sorting and co-assembly of aqueous supramolecular fibres were formed using peptide amphiphiles having immiscible hydrophobic tails.

DOI： 10.1039/d1cc05560g

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Scopus

PubMed

researchmap

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1246/cl.210231

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.aspen.2021.04.011

Language：Others   Publishing type：Research paper (scientific journal)  

The tumor necrosis factor α (TNFα) has been employed as a promising reagent in treating autoimmunity and cancer diseases. To meet the substantial requirement of TNFα proteins, we report in this study that mature types of recombinant human and murine TNFα proteins are successfully expressed in the baculovirus expression system using silkworm larvae as hosts. The biological activities of purified products were verified in culture murine L929 cells, showing better performance over a commercial Escherichia coli-derived murine TNFα. By comparing the activity of purified TNFα with or without the tag removal, it is also concluded that the overall activity of purified TNFα cytokines could be further improved by the complete removal of C-terminal fusion tags. Collectively, our current attempt demonstrates an alternative platform for supplying high-quality TNFα products with excellent activities for further pharmaceutical and clinical trials.

DOI： 10.3390/insects12060517

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/ijms22073459

Language：English  

DOI： https://doi.org/10.1101/2021.03.13.435221

Language：English   Publishing type：Research paper (scientific journal)  

DOI： https://doi.org/10.2116/analsci.20SCR07

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.2116/analsci.20scp12

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/ijms22073459

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.bioconjchem.1c00027

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ejps.2020.105667

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.pep.2020.105730

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/d0cc04491a

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.0c00598

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ab.2020.113700

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jbiosc.2020.05.010

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2020.119335

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2020.119335

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/pharmaceutics12040392

Language：Others  

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.aspen.2019.12.014

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/pharmaceutics12040392

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1038/s41598-020-63362-4

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.aspen.2019.12.014

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1248/bpb.b19-00642

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/pharmaceutics12030240

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acsomega.9b04163

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/pharmaceutics12030240

Language：English   Publishing type：Research paper (scientific journal)  

An allergy to cow’s milk proteins is the most common food allergy in infants and toddlers. Conventional oral immunotherapy for cow’s milk allergies requires hospital admission due to the risk of severe allergic reactions, including anaphylaxis. Therefore, a simpler and safer immunotherapeutic method is desirable. We examined transcutaneous immunotherapy with a solid-in-oil (S/O) system. In the S/O system, nano-sized particles of proteins are dispersed in an oil-vehicle with the assistance of nonionic surfactants. In the present study, the S/O system enhanced the skin permeation of the allergen molecule β-lactoglobulin (BLG), as compared with a control PBS solution. The patches containing BLG in the S/O nanodispersion skewed the immune response in the allergy model mice toward T helper type 1 immunity, indicating the amelioration of allergic symptoms. This effect was more pronounced when the immunomodulator resiquimod (R-848) was included in the S/O system.

DOI： 10.3390/pharmaceutics12030205

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.molliq.2019.112166

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.9b01104

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.9b01104

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2019.118777

Language：English   Publishing type：Research paper (scientific journal)  

Synthetic Strategies for Artificial Lipidation of Functional Proteins

DOI： 10.1002/chem.201904568

Language：Others  

Solid-in-oil nanodispersions for intranasal vaccination: Enhancement of mucosal and systemic immune responses.

DOI： 10.1016/j.ijpharm.2019.118777

Language：Others  

A novel binary supercooled liquid formulation for transdermal drug delivery.

DOI： 10.1248/bpb.b19-00642

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/pharmaceutics11110563

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2019.118582

Language：Others  

Transcutaneous Delivery of Immunomodulating Pollen Extract-Galactomannan Conjugate by Solid-in-Oil Nanodispersions for Pollinosis Immunotherapy.

DOI： 10.3390/pharmaceutics11110563

Language：English   Publishing type：Research paper (scientific journal)  

Ionic liquid (IL)surfactants have attracted great interest as promising substitutes for conventional surfactants owing to their exceptional and favorable physico-chemical properties. However, most IL surfactants are not eco-friendly and form unstable micelles, even when using a high concentration of the surfactant. In this study, we prepared a series of halogen-free and biocompatible choline–fatty-acid-based ILs with different chain lengths and degrees of saturation, and we then investigated their micellar properties in aqueous solutions. Characterization of the synthesized surface-active ILs (SAILs)was performed by ¹ H and ¹³ C nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, and elemental analysis. The surface-active properties of the SAILs were investigated by tensiometry, conductometry, and dynamic light scattering measurements. The critical micelle concentration of the SAILs was found to be 2–4 times lower than those of conventional surfactants. The thermodynamic properties of micellization (ΔG ⁰ _m , ΔH ⁰ _m , and ΔS ⁰ _m )indicate that the micellization process of the SAILs is spontaneous, stable, and entropy-driven at room temperature. The cytotoxicity of the SAILs was evaluated using mammalian cell line NIH 3T3. Importantly, [Cho][Ole]shows lower toxicity than the analogous ILs with conventional surfactants. These results clearly suggest that these environmentally friendly SAILs can be used as a potential alternative to conventional ILs for various purposes, including biological applications.

DOI： 10.1016/j.jcis.2019.04.095

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2019.118495

Language：English   Publishing type：Research paper (scientific journal)  

A nano-sized gel-in-oil suspension for transcutaneous protein delivery
We developed a new oil-based delivery system for transdermal protein delivery, a gel-in-oil (G/O) nanosuspension, where gelatin-based hydrogel was coated with hydrophobic surfactants. The high entrapment efficiency of a model protein, phycocyanin (PC), into nano-sized gelatin hydrogel particles was achieved. Spectroscopic evaluation of PC suggested that the G/O nanosuspension could retain the functional form of PC in isopropyl myristate. In vitro skin permeation studies showed that the G/O nanosuspension facilitated the delivery of PC through the stratum corneum of Yucatan micropig skin.

DOI： 10.1016/j.ijpharm.2019.118495

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.pep.2019.03.010

Language：Others  

Expression and purification of biologically active human granulocyte-macrophage colony stimulating factor (hGM-CSF) using silkworm-baculovirus expression vector system.

DOI： 10.1016/j.pep.2019.03.010

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2019.05.020

Language：English   Publishing type：Research paper (scientific journal)  

Polymerization of Horseradish Peroxidase by a Laccase-Catalyzed Tyrosine Coupling Reaction.
The polymerization of proteins can create newly active and large bio-macromolecular assemblies that exhibit unique functionalities depending on the properties of the building block proteins and the protein units in polymers. Herein, the first enzymatic polymerization of horseradish peroxidase (HRP) is reported. Recombinant HRPs fused with a tyrosine-tag (Y-tag) through a flexible linker at the N- and/or C-termini are expressed in silkworm, Bombyx mori. Trametes sp. laccase (TL) is used to activate the tyrosine of Y-tagged HRPs with molecular O2 to form a tyrosyl-free radical, which initiates the tyrosine coupling reaction between the HRP units. A covalent dityrosine linkage is also formed through a HRP-catalyzed self-crosslinking reaction in the presence of H2 O2 . The addition of H2 O2 in the self-polymerization of Y-tagged HRPs results in lower activity of the HRP polymers, whereas TL provides site-selectivity, mild reaction conditions and maintains the activity of the polymeric products. The cocrosslinking of Y-tagged HRPs and HRP-protein G (Y-HRP-pG) units catalyzed by TL shows a higher signal in enzyme-linked immunosorbent assay (ELISA) than the genetically pG-fused HRP, Y-HRP-pG, and its polymers. This new enzymatic polymerization of HRP promises to provide highly active and functionalized polymers for biomedical applications and diagnostics probes.

DOI： 10.1002/biot.201800531

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acsabm.9b00275

Language：Others  

Development of a novel ionic liquid-curcumin complex to enhance its solubility, stability, and activity

Language：Others  

Complementary interaction with peptide amphiphiles guides size-controlled assembly of small molecules for intracellular delivery

DOI： 10.1039/c9cc02473e

Language：English   Publishing type：Research paper (scientific journal)  

Rapid, convenient, sensitive detection methods are of the utmost importance in analytical tools. Enzyme-based signal amplification using horseradish peroxidase (HRP) is commonly implemented in clinical diagnostics kits based on enzyme-linked immunosorbent assay (ELISA), by which the limit of detection is greatly improved. Herein we report the design and preparation of recombinant fusion proteins comprising HRP and streptavidin (Stav), in which HRP was fused to either the N- or C-terminus of Stav ((HRP)
₄
–Stav or Stav–(HRP)
₄
, respectively) using a baculovirus-silkworm expression system. Both (HRP)
₄
–Stav and Stav–(HRP)
₄
were secreted in the apo form but they were easily converted to the holo form and activated by simple incubation with hemin overnight at 4 °C. The activated (HRP)
₄
–Stav and Stav–(HRP)
₄
could be combined with a commercial biotinylated anti-OVA IgG antibody to detect ovalbumin (OVA) as the antigen in ELISA. The enzymatic activity of (HRP)
₄
–Stav was twofold higher than that of Stav–(HRP)
₄
, and the sensitivity of (HRP)
₄
-Stav in ELISA was higher than that of a commercial HRP–Stav chemical conjugate. The successful use of (HRP)
₄
–Stav chimeric protein as a molecular probe in ELISA shows that the baculovirus-silkworm expression system is promising to produce enzyme–Stav conjugates to substitute for those prepared by chemical methods.

DOI： 10.1016/j.jbiotec.2019.03.007

Language：Others  

Language：English   Publishing type：Research paper (scientific journal)  

Molecular chaperone prefoldin-assisted biosynthesis of gold nanoparticles with improved size distribution and dispersion
Here we report a novel aspect of molecular chaperone prefoldin (PFD) as a biomaterial in the biocatalytic synthesis of gold nanoparticles (AuNPs) using glycerol dehydrogenase (GLD). We found that PFD could inhibit the aggregation of AuNPs during the biosynthesis, leading to the formation of AuNPs with controlled size distribution.

DOI： 10.1039/c8bm01026a

Language：Others  

In vivo biocompatibility, pharmacokinetics, antitumor efficacy, and hypersensitivity evaluation of ionic liquid-mediated paclitaxel formulations

Language：Others  

Functional horseradish peroxidase-streptavidin chimeric proteins prepared using a silkworm-baculovirus expression system for diagnostic purposes.

DOI： 10.1016/j.jbiotec.2019.03.007

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/chem.201900370

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1088/1755-1315/278/1/012077

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c8bm01026a

Language：Others  

Transcutaneous Codelivery of Tumor Antigen and Resiquimod in Solid-in-Oil Nanodispersions Promotes Antitumor Immunity

Language：Japanese  

Language：Others  

Self-Assembled Reduced Albumin and Glycol Chitosan Nanoparticles for Paclitaxel Delivery

DOI： 10.1021/acs.langmuir.8b02809

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jphotochem.2018.10.006

Language：Others  

Enhanced Potential of Therapeutic Applications of Curcumin Using Solid-in-Water Nanodispersion Technique

DOI： 10.1252/jcej.18we060

Language：Others  

Effect of macromolecular crowding on the conformational behaviour of a porphyrin rotor

DOI： 10.1016/j.jphotochem.2018.10.006

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acsbiomaterials.9b00260

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c9cc02812a

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c9cc02473e

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201800531

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.18we060

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acsabm.8b00271

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201800085

Language：Others  

Designer aromatic peptide amphiphiles for self-assembly and enzymatic display of proteins with morphology control

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jbiosc.2018.05.013

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jbiosc.2018.05.013

Language：Others  

Design of lipid-protein conjugate with a self-assembling ability on a cell membrane by using microbial transglutaminase reaction

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.aspen.2018.05.002

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.aspen.2018.05.002

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ejpb.2018.01.020

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.8b00305

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201700624

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.8b00305

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.colsurfb.2018.01.041

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.procbio.2017.12.013

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.7b00894

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.procbio.2017.12.013

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.molpharmaceut.7b00894

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.17we293

Language：Others  

DOI： 10.1002/9783527803293.ch6

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.11416/jibs.87.2_053

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201700195

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.bioconjchem.7b00594

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2017.07.020

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2017.07.020

Language：English  

DOI： 10.1252/jcej.16we335

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s12010-016-2322-2

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00726-016-2322-0

Language：English   Publishing type：Research paper (scientific journal)  

Transcutaneous pollinosis immunotherapy using a solid-in-oil nanodispersion system carrying T cell epitope peptide and R848.
Antigen-specific immunotherapy is the only curative approach for the treatment of allergic diseases such as Japanese cedar pollinosis. Immunotherapy using a T cell epitope vaccine in combination with the adjuvant R848 is of particular interest as a safe and effective approach to treat allergic diseases. Herein, we propose a simple and easy to handle vaccine administration method using the original solid-in-oil (S/O) nanodispersion system that permeates through the skin. The S/O nanodispersion system is composed of nanoparticles of hydrophilic molecules surrounded with hydrophobic surfactants that are dispersed in an oil vehicle. The system has potential to carry and deliver both hydrophilic and hydrophobic bioactives. Hydrophilic T cell epitope peptide was efficiently delivered through mouse skin using the S/O nanodispersion system and lowered antigen-specific IgE levels in pollinosis model mice. Addition of the hydrophobic adju1vant R848 significantly lowered the antibody secretion and shifted the Th1/Th2-balance toward Th1-type immunity in the model mice, showing the potential to alleviate Japanese cedar pollinosis.

DOI： 10.1002/btm2.10048

Language：English  

DOI： 10.1007/s00726-016-2322-0

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.biomac.6b01538

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.biomac.6b01538

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.16we335

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s12010-016-2322-2

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00726-016-2322-0

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201600081

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201600081

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201600087

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201600650

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2016.07.013

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2016.07.013

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/acs.biomac.6b01141

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2016.06.005

Language：Others  

Diglycolic amic acid-modified E. coli as a biosorbent for the recovery of rare earth elements
© 2016 Elsevier B.V. Biosorption has recently attracted much attention as an alternative to conventional techniques for the recovery of rare earth elements (REEs). In this study, Escherichia coli (E. coli) was chemically modified to improve its performance as a biosorbent for REEs. The diglycolic amic acid group, which shows high affinity to REEs, was introduced by succinylation of the amine groups on the E. coli. Adsorption curves using the modified E. coli were characteristic of the diglycolic amic acid group. The adsorption performance for transition metal ions was not affected by the modification. These results suggest that modification of E. coli with a functional group with high affinity to REEs increases the effectiveness of adsorption. The maximum uptakes of REEs on the modified E. coli were doubled. Modification of E. coli is an effective method for enhancing the adsorption performance for REEs.

DOI： 10.1016/j.bej.2016.06.005

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/C6RA16926K

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2016.04.121

Language：Others  

Powerful peracetic acid-ionic liquid pretreatment process for the efficient chemical hydrolysis of lignocellulosic biomass
© 2016 Elsevier Ltd. The aim of this work was to design a new method for the efficient saccharification of lignocellulosic biomass (LB) using a combination of peracetic acid (PAA) pretreatment with ionic liquid (IL)-HCl hydrolysis. The pretreatment of LBs with PAA disrupted the lignin fractions, enhanced the dissolution of LB and led to a significant increase in the initial rate of the IL-HCl hydrolysis. The pretreatment of Bagasse with PAA prior to its 1-buthyl-3-methylimidazolium chloride ([Bmim][Cl])-HCl hydrolysis, led to an improvement in the cellulose conversion from 20% to 70% in 1.5 h. Interestingly, the 1-buthyl-3-methylpyridium chloride ([Bmpy][Cl])-HCl hydrolysis of Bagasse gave a cellulose conversion greater than 80%, with or without the PAA pretreatment. For LB derived from seaweed waste, the cellulose conversion reached 98% in 1 h. The strong hydrolysis power of [Bmpy][Cl] was attributed to its ability to transform cellulose I to II, and lowering the degree of polymerization of cellulose.

DOI： 10.1016/j.biortech.2016.04.121

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/C6OB01227B

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.aspen.2016.03.014

Language：Others  

High-level expression and purification of biologically active human IL-2 using silkworm-baculovirus expression vector system

DOI： 10.1016/j.aspen.2016.03.014

Language：Others  

Enzymatic conjugation of multiple proteins on a DNA aptamer in a tail-specific manner
Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Conjugation of single-strand DNA aptamers and enzymes has been of great significance in bioanalytical and biomedical applications because of the unlimited functions provided by DNA aptamer direction. Therefore, we developed efficient tailing of a DNA aptamer, with end-specific conjugation of multiple enzymes, through enzymatic catalysis. Terminal deoxynucleotidyl transferase (TdT) added multiple Z-Gln-Gly (Z-QG) moieties to the 3′-end of a DNA aptamer via the addition of Z-QG-modified deoxyuridine triphosphate (Z-QG-dUTP) and deoxynucleoside triphosphates (dNTPs). The resultant (Z-QG)m-(dN)l-aptamer, whose Z-QGs with dN spacers served as stickers for microbial transglutaminase (MTG), were crosslinked between the Z-QGs on the DNA and a substrate peptide sequence containing lysine (K), fused to a recombinant enzyme (i.e. bacterial alkaline phosphatase; BAP) by MTG. The incorporation efficiency of Z-QG moieties on the aptamer tail and the subsequent conjugation efficiency with multiple enzyme molecules were dramatically altered by the presence of dNTPs, revealing that a combination of Z-QG-dUTP/dTTP comprised the best labeling efficiency and corresponding properties during analytical performance. Thus, a novel optimized platform for designing (BAP)n-(dT)l-DNA aptamers was demonstrated for the first time in this article, offering unique opportunities for tailoring new types of covalent protein-nucleic acid conjugates in a controllable way.

DOI： 10.1002/biot.201500560

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biot.201500560

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c6ra16926k

Language：English  

DOI： 10.1007/978-4-431-55825-5_17

Language：Others  

© The Royal Society of Chemistry 2016. β-Annulus peptides bearing Cys at the N-terminal from tomato bushy stunt virus were synthesised using a standard Fmoc-protected solid-phase method, and the peptide was modified with Ni-NTA at the N-terminal. The Ni-NTA-modified β-annulus peptide self-assembled into virus-like nanocapsules of approximately 40 nm in diameter. The critical aggregation concentration of these nanocapsules in 10 mM Tris-HCl buffer (pH 7.3) at 25 °C was 0.053 μM, which is 470 times lower than that of unmodified β-annulus peptides. Moreover, size exclusion chromatography of the peptide assembly indicated encapsulation of His-tagged green fluorescent protein in the Ni-NTA-modified artificial viral capsid.

DOI： 10.1039/c6ob01227b

Language：Others  

Selective extraction of scandium from transition metals by synergistic extraction with 2-thenoyltrifluoroacetone and tri-n-octylphosphine oxide
There has been an increasing demand for scandium (Sc). However the supply is insufficient because the separation and recovery of Sc as a byproduct from ores is difficult. In this study, extraction and separation of Sc3+from a sulfuric acid solution containing transition metals such as Al3+, Fe3+, Mn2+, Co2+, Ni2+and Zn2+was investigated using a β-diketone, HTTA and a neutral extractant, TOPO. Due to the synergistic effect with the two extractants, Sc3+was effectively and selectively separated even from Fe3+, the separation of which is difficult with HTTA alone. The extraction mechanism was examined and it was revealed that Sc3+is extracted with three HTTA and one TOPO molecules. Scandium was readily stripped from the extracting phase by 1 M sulfuric acid. The potential use of the combination of the β-diketone with TOPO for the recovery of Sc3+in a refining process was demonstrated.

DOI： 10.15261/serdj.23.137

Language：Others  

Mutual separation of indium, gallium, and zinc with the amic acid-type extractant D2EHAG containing glycine and amide moieties
Solvent extraction of indium (In3+), gallium (Ga3+), and zinc (Zn2+) was investigated using N-[N,N-di(2-ethylhexyl)aminocarbonylmethyl]glycine (D2EHAG) and N-[N,N-di(2-ethylhexyl)aminocarbonylmethyl]-sarcosine (D2EHAS) which we have developed recently. Indium and gallium were selectively extracted from zinc under high acid conditions (0 > pH ≥ 2.0), and easily stripped using an acidic solution such as 2 mol dm-3 HNO3. The extraction behavior was compared to that of N,N-dioctyldiglycol amic acid (DODGAA), which has a similar molecular structure to D2EHAG, a commercial alkyl monocarboxylic acid extractant, neodecanoic acid (Versatic 10), or an organophosphorus extractant, di(2-ethylhexyl) phosphoric acid (D2EHPA). Based on the results, D2EHAG and D2EHAS were found to be useful for the separation of In3+ and Ga3+ from Zn2+. The extraction mechanisms of these ions with D2EHAG were investigated through slope analysis and loading tests.

DOI： 10.15261/serdj.23.9

Language：Others  

Biocatalytic formation of gold nanoparticles decorated with functional proteins inside recombinant Escherichia coli cells
© The Japan Society for Analytical Chemistry. A novel strategy for the preparation of protein-decorated gold nanoparticles (Au NPs) was developed inside Escherichia coli cells, where an artificial oxidoreductase, composed of antibody-binding protein (pG), Bacillus stearothermophilus glycerol dehydrogenase (BsGLD) and a peptide tag with gold-binding affinity (H6C), was overexpressed in the cytoplasm. In situ formation of Au NPs was promoted by a natural electron-donating cofactor, nicotinamide adenine dinucleotide (NAD), which was regenerated to the reduced form of NADH by the catalytic activity of the fusion protein (pG-BsGLDH6C) overexpressed in the cytoplasm of E. coli, with the concomitant addition of exogenous glycerol to the reaction system. The fusion protein was self-immobilized on Au NPs inside the E. coli cells, which was confirmed by SDS-PAGE and western blotting analyses of the resultant Au NPs. Finally, the IgG binding ability of the pG moiety displayed on Au NPs was evaluated by an enzyme-linked immunosorbent assay. 2016

DOI： 10.2116/analsci.32.295

Language：Others  

BODIPY-labeled fluorescent aptamer sensors for turn-on sensing of interferon-gamma and adenine compounds on cells
© The Japan Society for Analytical Chemistry. An on-cell aptamer sensor has the potential to reveal cell-cell communications by signalling molecules. We attempted to design new fluorescent aptamer sensors for the sensing of IFN-? and adenine compounds on cells. BODIPY-labeled external quencher-free aptamer sensors have allowed a turn-on detection of the target molecule with improved off/on efficiency.

DOI： 10.2116/analsci.32.543

Language：English  

DOI： 10.1007/978-4-431-55825-5_17

Language：English   Publishing type：Research paper (scientific journal)  

β-Annulus peptides bearing Cys at the N-terminal from tomato bushy stunt virus were synthesised using a standard Fmoc-protected solid-phase method, and the peptide was modified with Ni-NTA at the N-terminal. The Ni-NTA-modified β-annulus peptide self-assembled into virus-like nanocapsules of approximately 40 nm in diameter. The critical aggregation concentration of these nanocapsules in 10 mM Tris-HCl buffer (pH 7.3) at 25 °C was 0.053 μM, which is 470 times lower than that of unmodified β-annulus peptides. Moreover, size exclusion chromatography of the peptide assembly indicated encapsulation of His-tagged green fluorescent protein in the Ni-NTA-modified artificial viral capsid.

DOI： 10.1039/c6ob01227b

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.2116/analsci.32.543

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.23.137

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c5me00011d

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.23.9

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.2116/analsci.32.295

Language：Japanese   Publishing type：Research paper (scientific journal)  

DOI： 10.1208/s12249-015-0333-x

Language：English  

Transcutaneous Peptide Immunotherapy of Japanese Cedar Pollinosis Using Solid-in-Oil Nanodispersion Technology
© 2015, American Association of Pharmaceutical Scientists. Peptide immunotherapy is an attractive approach to relieve allergic symptoms such as rhinitis and asthma. Treatment of Japanse cedar pollinosis (Cryptomeria japonica; Cj), from which over one quarter of Japanese population suffer, is becoming a great concern. Recently, oral feeding of a peptide (7crp) consisting of seven immunodominant human T cell epitopes derived from two enzymes present in Cj pollen was demonstrated to have a benefit in treating Cj pollinosis. In this work, we aimed to apply a novel transcutaneous administration system as a simple and easy peptide delivery for an immunotherapy using a T cell epitope peptide. A modified 7crp peptide (7crpR) which contained triarginine linkers between each epitopes was designed to increase water solubility and was encapsulated in a unique solid-in-oil (S/O) nanodispersion. The S/O nanodispersion consists of a nano-sized peptide-surfactant complex dispersed in an oil vehicle. The S/O nanopartilces having an average diameter of 230 nm facilitated the permeation of the peptide 7crpR into the skin and suppressed serum total IgE and antigen-specific IgE levels in a Cj pollinosis mouse model. Transcutaneous administration of the T cell epitope peptide using the S/O nanodispersion system has potential for future simple and easy immunotherapy of Cj pollinosis.

DOI： 10.1208/s12249-015-0333-x

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1246/cl.150794

Language：English   Publishing type：Research paper (scientific journal)  

Language：Japanese   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biombioe.2015.05.023

Language：Others  

Heme precursor injection is effective for Arthromyces ramosus peroxidase fusion protein production by a silkworm expression system

DOI： 10.1016/j.jbiosc.2015.02.013

Language：Others  

Great potency of seaweed waste biomass from the carrageenan industry for bioethanol production by peracetic acid-ionic liquid pretreatment
© 2015 Published by Elsevier Ltd. Seaweed waste biomass from the carrageenan industry (SWBC) is a potential biomass feedstock for producing sustainable biofuel because it increases the product value and reduces the pollutant risk. Peracetic acid (PAA) followed by ionic liquid (IL) pretreatment has been used to increase the enzymatic saccharification of pretreated SWBC. The SWBC cellulose content was comparable with that of terrestrial biomasses. PAA+1-hexylpyridinium chloride ([Hpy][Cl]), and PAA+1-ethyl-3-methylimidazolium diethylphosphate ([Emim][DEP]) pretreatments produced saccharide and unknown oligosaccharide fractions in regenerated water (~4-6% SWBC cellulose content). For 48h of saccharification, the untreated SWBC and the SWBC pretreated using PAA followed by [Hpy][Cl], [Emim][DEP] or 1-ethyl-3-methylimidazole acetate ([Emim][OAc]) produced cellulose conversions of 77, 91, 84 and 62%, respectively. The untreated SWBC had a high cellulose conversion, which may be caused by the low lignin and hemicellulose contents of the SWBC. PAA+IL pretreatment could yield pretreated SWBCs with more amorphous cellulose structures, which lead to an almost-complete cellulose conversion.

DOI： 10.1016/j.biombioe.2015.05.023

Language：Japanese   Publishing type：Research paper (scientific journal)  

Language：Others  

Transcutaneous immunization against cancer using solid-in-oil nanodispersions
© The Royal Society of Chemistry. Transcutaneous immunization is a novel, non-invasive alternative to conventional immunization by injection. Skin immunocompetence composed of abundant antigen-presenting cells in the epidermal and dermal layers of the skin can provide an effective tool for transcutaneous immunization, whereas the outermost, hydrophobic layer of skin, the stratum corneum, hinders the penetration of antigens into the skin. To realize effective transcutaneous delivery of antigens, we have developed a solid-in-oil (S/O) technique that produces an oil dispersion of hydrophilic biomolecules. In this study, we applied the S/O nanodispersions in transcutaneous immunization to induce cancer immunity. The topical application of S/O nanodispersions bearing ovalbumin (OVA) as a model cancer-antigen allowed the penetration of OVA into the deep dermis region of the skin by intercellular and transfollicular pathways. Inhibition of OVA-bearing tumor growth and production of cytokines responsible for cellular immunity were achieved, demonstrating the applicability of S/O nanodispersions to the induction of cancer immunity.

DOI： 10.1039/c5md00168d

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c5md00168d

Language：English   Publishing type：Research paper (scientific journal)  

Effect of pretreatment methods on the synergism of cellulase and xylanase during the hydrolysis of bagasse

DOI： 10.1016/j.biortech.2015.02.041

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.enzmictec.2015.01.007

Language：Others  

Synergistic effect and application of xylanases as accessory enzymes to enhance the hydrolysis of pretreated bagasse.

DOI： 10.1016/j.enzmictec.2015.01.007

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s11095-014-1554-5

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/kakoronbunshu.41.157

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.14we162

Language：Japanese   Publishing type：Research paper (scientific journal)  

Language：English  

DOI： 10.1007/978-1-4939-2303-8_29

Language：English  

DOI： 10.1007/978-1-4939-2303-8_29

Language：Japanese   Publishing type：Research paper (scientific journal)  

Language：Others  

Transdermal immunization using solid-in-oil nanodispersion with CpG oligodeoxynucleotide adjuvants
© 2014 Springer Science+Business Media New York. Purpose: Simple and noninvasive vaccine administration alternatives to injections are desired. A solid-in-oil (S/O) nanodispersion system was able to overcome skin barriers and induce an immune response; however, antibody levels remained low. We applied an immune potentiator, CpG oligodeoxynucleotide (ODN), to enhance the immune response by controlling the T helper 1 (Th1)/T helper 2 (Th2) balance. Methods: S/O nanodispersions containing ovalbumin (OVA) and CpG ODN (CpG-A or CpG-B) were characterized by size distribution analysis and a protein release test. The skin permeation of fluorescence-labeled OVA was observed by fluorescence microscopy. Antigen-specific IgG, IgG1, and IgG2a responses were measured by enzyme-linked immunosorbent assay. Results: Co-encapsulation of CpG ODNs in S/O nanodispersions enhanced induction of OVA-specific IgG. S/O nanodispersion containing OVA and CpG-A had a smaller mean particle size and permeated the skin more efficiently. In contrast, CpG-B showed the highest protein release and induction of OVA-specific IgG. IgG subclass analysis revealed that OVA induced a Th2-dominant immune response, while the S/O nanodispersion containing CpG-A skewed the immune response toward a Th1-bias. Conclusions: In combination with CpG ODN, the S/O nanodispersion system efficiently induced an antigen-specific antibody response. The Th1/Th2 immune balance could be controlled by the selection of CpG ODN type.

DOI： 10.1007/s11095-014-1554-5

Language：Others  

Site-specific conjugation of an antibody-binding protein catalyzed by horseradish peroxidase creates a multivalent protein conjugate with high affinity to IgG
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Cross-linking proteins offers an approach to enhance the distinct function of proteins due to the multivalent effect. In this study, we demonstrated the preparation of a multivalent antibody-binding protein possessing high affinity to IgG by conjugating a number of antibody-binding proteins using the horseradish peroxidase (HRP)-mediated protein conjugation method. By introducing a peptide tag containing a tyrosine (Y-tag) to the C-terminus of the model protein, a chimera protein of protein G and protein A (pG2pA), the Tyr residue in the Y-tag was efficiently recognized by HRP and cross-linked with each other to yield a pG2pA conjugate, composed of mainly two to three units of pG2pA. The cross-linking occurred site specifically at the Tyr residue in the Y-tag and introduction of the Y-tag showed no effect on the function of pG2pA. The affinity of the Y-tagged pG2pA conjugate against IgG clearly increased because of the multivalent effect, demonstrating the benefit of this protein cross-linking reaction, which yields functional protein oligomers. Such multivalent protein conjugates created by this reaction should have potential to be used in ELISA and Western blotting applications in which highly sensitive detection of target molecules is desired.

DOI： 10.1002/biot.201400512

Language：Others  

Separation of gold(III) in acidic chloride solution using porous polymeric ionic liquid gel
© 2015 The Society of Chemical Engineers, Japan. A novel polymeric ionic liquid (PIL) gel was synthesized via radical polymerization of the ionic liquid monomers, 1-vinyl- 3-ethylimidazoliumbis(trifluorometyl-sulfonyl) imide ([veim][Tf2N]). The gel was made porous by removing the water phase dispersed by polyoxyethylene (20) sorbitan monolaurate (Tween-20). The porous structure of the synthesized gel was observed by SEM imaging. In comparison to the nonporous PIL gel synthesized without Tween-20, the adsorption capacity of Au(III) onto porous PIL gel was enhanced. The adsorption behavior of the synthesized gel was studied in acidic chloride solutions containing various metal ions, and it was found that the gel selectively adsorbed Au(III) over other metal ions. Maximum adsorption capacity of Au(III) was more than 140 mg/g; whereas, those for Pt(IV) and Pd(II) were lower than 20 mg/g. The adsorbed Au(III) onto the gel can easily be desorbed using an acidified thiourea aqueous solution. The feasibility of utilizing the polymeric ionic liquid gel for the separation of Au(III) was thereby demonstrated.

DOI： 10.1252/jcej.14we162

Language：Others  

Ionic liquid-mediated transcutaneous protein delivery with solid-in-oil nanodispersions
© 2015 The Royal Society of Chemistry. As a potentially safe and non-invasive vaccination method, transcutaneous immunization represents an attractive alternative to conventional vaccine delivery by injection. However, the development of transcutaneous immunization has remained a challenge for a large number of hydrophilic macromolecules including protein and peptide antigens. We report a novel ionic liquid (IL)-mediated transcutaneous vaccine formulation consisting of a solid-in-oil (S/O) nanodispersion of antigen coated with pharmaceutically accepted surfactants dispersed in IL-containing oil. The introduction of the IL [C12mim][Tf2N] (1-dodecyl-3-methyl imidazolium bis(trifluoromethyl sulfonyl) amide) as a penetration enhancer in the formulation significantly enhanced the skin permeability of ovalbumin (OVA), a model antigen. It was also found that the IL-mediated S/O nanodispersion obtained high levels of OVA-specific serum IgG compared with both S/O nanodispersions without IL and PBS control. These findings clearly indicate that ILs-which are potentially attractive "green" and "designer" solvents-could serve as potential skin penetration enhancers in transcutaneous vaccination for hydrophilic macromolecules.

DOI： 10.1039/c5md00378d

Language：Others  

Enzyme-mediated preparation of hydrogels composed of poly(ethylene glycol) and gelatin as cell culture platforms
© The Royal Society of Chemistry 2015. A redox-responsive hydrogel composed of poly(ethylene glycol) and gelatin was prepared via an enzymatic oxidative reaction. Fibroblast cells adhered on the hydrogel showed proliferation and they could be recovered as a cell sheet by degradation of the scaffold under mild reductive conditions, thus providing a novel platform for cell culture.

DOI： 10.1039/c4ra12334d

Language：Others  

Activity and stability of enzyme immobilized with ionic liquid based polymer materials
© School of Engineering, Taylor’s University. Enzyme immobilization methods are continuously being developed for a wide range of applications including biocatalysis and biotransformation. Compared to other immobilization methods, physical entrapment into a matrix is relatively simple and inexpensive, and causes a relatively small perturbation to the native enzyme structure and function. However, significant enzyme leaching from such biocatalytic polymers is the main constraint in using them for industrial levels. This study reports an ionic liquid (IL) polymer materials incorporating enzymes that can be used as active, stable and reusable biocatalysts to overcome the limitations. Lipase was microencapsulated in surfactant aggregates formed in an IL monomer or the solution of an IL monomer/IL and then incorporated into polymer frameworks through the free radical polymerization of an IL (1-vinyl-3-ethylimidazolium bis(trifluoromethylsulfonyl) amide ([veim][Tf2N]). The activity, stability and reusability of IL polymer materials containing lipase were evaluated using lipase-catalyzed hydrolysis of p-nitrophenyl butyrate (p-PNB) as a model reaction. It was found that lipase IL polymer materials exhibited excellent stability in aqueous solutions. More importantly, these biopolymer materials retained most of their activity after six reaction cycles. We firmly believe that enzyme containing IL polymeric materials developed in this study will provide several advantages over conventional polymer based methods for diverse enzymatic reactions.

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/978-1-4939-2303-8_29

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/kakoronbunshu.41.38

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1246/cl.150794

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c5md00378d

Language：Others   Publishing type：Research paper (scientific journal)  

DOI： 10.1186/s40508-014-0019-9

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c4md00179f

Language：Others  

Application of cellulose acetate to the selective adsorption and recovery of Au(III)
Cellulose acetyl derivatives were examined for the selective recovery of Au(III) from acidic chloride solutions as an adsorbent, and cellulose acetate fibers (CAF) were found to be effective for the separation of Au(III) from other metal ions, including the precious metal ions Pt(IV) and Pd(II). The amount of Au(III) adsorbed by the fibers increased with an increase in the hydrochloric acid concentration, but decreased with an increase in the ionic strength of the solution. The adsorption of Au(III) onto CAF took place quickly and an adsorption equilibrium was reached within 1 h. The maximum adsorption capacity of Au(III) was determined to be 110 mg/g at 2 M hydrochloric acid. The loaded Au(III) was readily recovered by incineration. © 2014 Elsevier Ltd.

DOI： 10.1016/j.carbpol.2014.05.003

Language：Others  

A novel surface-coated nanocarrier for efficient encapsulation and delivery of camptothecin to cells
© 2014 the Partner Organisations. In the present study, we developed a novel surface-coated nanocarrier (SCN) for efficient and stable encapsulation of a poorly water-soluble anticancer agent, camptothecin (CPT). Using emulsification and freeze-drying processes in the presence of a hydrophilic surfactant, Pluronic F-68 or F-127, CPT-Pluronic complexes with crystalline features were synthesised. Investigation of the in vitro anticancer activities and cellular internalisation of the complexes revealed that the SCN efficiently delivered CPT to cells while maintaining anticancer activity.

DOI： 10.1039/c4md00179f

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.carbpol.2014.05.003

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2014.09.003

Language：Others  

Characterization of enzymatically gellable, phenolated linear poly(ethylene glycol) with different molecular weights for encapsulating living cells
© 2014 Elsevier B.V. Enzymatic hydrogelation has received much attention due to the high biocompatibility and the ease of control of reaction kinetics under physiological conditions. In particular, horseradish peroxidase (HRP)-mediated phenol coupling reaction has great potential for developing in situ hydrogelation systems. Herein, we report the HRP-catalyzed preparation and characterization of hydrogels composed of a terminally bis-phenolated linear poly(ethylene glycol) (PEG-Ph-OH) with different molecular weights (Mws 3100, 8800, 11,000, 20,000g/mol). The gelation time of polymer solution can be controlled in the range from few second to few minute, suggestion that the PEG-Ph-OH has a potential as a in situ forming hydrogel. In addition, the physicochemical properties of the hydrogels, such as swelling ratio, mesh size and mechanical property, were controlled by the molecular weight of the PEG-Ph-OH. The results could be attributed to the alteration in the cross-linking density by the variation of molecular weight of the gel precursor. Furthermore, the viability of mammalian cells encapsulated in the PEG-Ph-OH hydrogels was approximately 90%. These results indicate that PEG-Ph-OH has potential for biomedical applications including tissue engineering.

DOI： 10.1016/j.bej.2014.09.003

Language：English  

DOI： 10.1252/jcej.14we360

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/ie403524a

Language：Others  

The self-assembly and secondary structure of peptide amphiphiles determine the membrane permeation activity
Membrane fusogenic peptides have attracted increasing attention because of their unique biofunctions in membrane translocation and viral infection. Here, we designed GALA-related peptides with palmitoyl tails. Our study indicated that the self-assembling propensity and the secondary structure of these peptide amphiphiles greatly influenced the membrane permeability. This journal is © the Partner Organisations 2014.

DOI： 10.1039/c4ra02901a

Language：Others  

Sucrose laurate enhanced transcutaneous immunization with a solid-in-oil nanodispersion

DOI： 10.1039/c3md00164d

Language：Others  

Separation of precious metals by using undiluted ionic liquids
The ionic liquid (IL), 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([Bmim][Tf2N]), was evaluated as an extractant without dilution for the separation and recovery of precious metals. Au(III) was efficiently extracted from the aqueous phase to the IL phase, and a concentrated Au(III) solution almost 8 times higher than the initial feed solutions could be produced. In contrast, Pd(II) was rarely extracted and Pt(IV) was partly extracted under the present experimental conditions. Furthermore, the effect of the IL anions on the extraction ability for the metal ions was examined, and it was found that an IL having an anion component slightly more hydrophilic compared to [Tf2N], such as 1-butyl-3-methylimidazolium hexafluorophosphate [Bmim][PF6], showed a higher extraction ability than [Bmim][Tf2N]. It was suggested that the extraction proceeded via an anion exchange mechanism and it was shown that ILs could be effective extractants for the separation of precious metals.

DOI： 10.15261/serdj.21.89

Language：Others  

One step effective separation of platinum and palladium in an acidic chloride solution by using undiluted ionic liquids
Imidazolium-based ionic liquids (ILs) with Tf2N- as anions were studied as extractants without dilution for the extractive separation of platinum and palladium in an acidic chloride solution. The effect of the alkyl chain length on the extraction performance was evaluated and, based on the performance, the IL, 1-methyl-3-octylimidazolium bis(trifluoromethylsulfonyl) imide ([Omim][Tf2N]), was selected to be studied in detail. The extraction of Pt(IV) underwent little change over the HCl concentration studied, while that of Pd(II) decreased with increasing HCl concentration. The separation of Pt(IV) and Pd(II) was highly effective, and the highest separation factor between Pt(IV) and Pd(II) reached 312 at the optimum conditions in a binary component system composed of Pt(IV) and Pd(II). The effect of the initial Pt(IV) concentration on loading capacity and extraction efficiency of Pt(IV) was also investigated.

DOI： 10.15261/serdj.21.129

Language：Others  

Development of novel extractants with amino acid structure for efficient separation of nickel and cobalt from manganese ions
The solvent extraction separation of nickel and cobalt from manganese is an important issue to be resolved. Novel amic acid type extractants with the glycine or sarcosine moiety N-[N,N-di(2-ethylhexyl)aminocarbonylmethyl]glycine (D2EHAG) and N-[N,N-di(2-ethylhexyl)aminocarbonylmethyl]sarcosine (D2EHAS) were synthesized to achieve this separation. These extractants were prepared by a simple two-step reaction and were readily soluble in aliphatic organic solvents. The extraction behavior of Ni2+, Co2+, and Mn 2+ with D2EHAG and D2EHAS in n-dodecane was investigated compared with that of conventional extractants. The novel extractants extracted Ni 2+ and Co2+ preferentially with a high selectivity to Mn2+. The metal ions could be back-extracted quantitatively using 1 mol dm-3 sulfuric acid. The extraction mechanism of the three metal ions with D2EHAG and D2EHAS was investigated, and results suggest that one divalent metal ion is extracted with two extractant molecules. © 2013 American Chemical Society.

DOI： 10.1021/ie403524a

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c4ra08897b

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c3ra45668d

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c3md00164d

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.21.129

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c4ra02901a

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.14we360

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.21.89

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jbiosc.2013.05.025

Language：Others  

Tailing DNA aptamers with a functional protein by two-step enzymatic reaction
An efficient, quantitative synthetic strategy for aptamer-enzyme conjugates was developed by using a two-step enzymatic reaction. Terminal deoxynucleotidyl transferase (TdT) was used to first incorporate a Z-Gln-Gly (QG) modified nucleotide which can act as a glutamine donor for a subsequent enzymatic reaction, to the 3&#039;-OH of a DNA aptamer. Microbial transglutaminase (MTG) then catalyzed the cross-linking between the Z-QG modified aptamers and an enzyme tagged with an MTG-reactive lysine containing peptide. The use of a Z-QG modified dideoxynucleotide (Z-QG-ddUTP) or a deoxyuridine triphosphate (Z-QG-dUTP) in the TdT reaction enables the controlled introduction of a single or multiple MTG reactive residues. This leads to the preparation of enzyme-aptamer and (enzyme)n-aptamer conjugates with different detection limits of thrombin, a model analyte, in a sandwich enzyme-linked aptamer assay (ELAA). Since the combination of two enzymatic reactions yields high site-specificity and requires only short peptide substrates, the methodology should be useful for the labeling of DNA/RNA aptamers with proteins. © 2013 The Society for Biotechnology, Japan.

DOI： 10.1016/j.jbiosc.2013.05.025

Language：Others  

Needle-free immunization using a solid-in-oil nanodispersion enhanced by skin-permeable oligoarginine peptide

DOI： 10.1016/j.ijpharm.2013.10.006

Language：Others  

Enzymatic fabrication of protein-decorated gold nanoparticles by the aid of artificial peptides with gold-binding affinity
Here, we report a new approach for the biofabrication of protein-immobilized gold nanoparticles (Au NPs), using oxidoreductase with gold-binding peptide-tagged recombinant proteins. The reduction of Au ions to Au0 was achieved using a natural electron-donating cofactor, nicotinamide adenine dinucleotide, which was regenerated by the glycerol dehydrogenase (GLD) enzyme. First, we selected the A3 peptide (AYSSGAPPMPPF) as a gold binding moiety. The A3 peptide was introduced to the C-terminus of fusion proteins of immunoglobulin G (IgG)-binding domains of protein G and protein A. In the presence of the recombinant protein, the GLD-catalyzed cofactor reduction resulted in the efficient in situ fabrication of Au NPs immobilized with the fusion protein. Moreover, the protein-immobilized Au NPs were shown to have IgG binding activity. Although the A3 peptide had the ability to stabilize Au NPs, the results suggested that its binding affinity for Au NPs was unexpectedly weaker than that of His-tag. A cysteine residue was thus introduced to a recombinant protein adjacent to the A3 peptide. Finally, an artificial peptide, comprising A3 sequence with the C-terminal single cysteine residue, enabled the stable display of a fusion protein while maintaining its IgG binding activity through the Au-S bond. This enzyme-assisted one-pot methodology for protein-Au NPs conjugation offers one potent route for the facile fabrication of biomolecule-decorated metal NPs. © 2013 American Chemical Society.

DOI： 10.1021/la401327h

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2013.10.006

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/la401327h

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.13we009

Language：Others  

Ionic liquid-in-oil microemulsions as potential carriers for the transdermal delivery of Methotrexate
Ionic liquid-in-oil (IL/o) microemulsion (ME) techniques have been employed to increase the solubility of sparingly soluble drugs and enhance the extent of their topical and transdermal delivery. Methotrexate (MTX) has been used as a model drug in the current study. The size and size distributions of the ME droplets in the presence and in the absence of MTX were determined by dynamic light scattering analysis. The mean droplet diameter of the ME droplets containing 4 mg/mL of MTX was less than 20 nm, and smaller than the corresponding value observed in the absence of MTX. MTX was incorporated into IL droplets as well as other formulations and its permeation across Yucatan micropig (YMP) porcine skin evaluated. The use of IL/o ME has been shown to dramatically increase MTX administration. The results obtained in this study reveal the versatility of IL/o MEs as efficient drug delivery systems for sparingly soluble drugs. © 2013 The Society of Chemical Engineers, Japan.

DOI： 10.1252/jcej.13we009

Language：Others  

Aligning an endoglucanase Cel5A from Thermobifida fusca on a DNA scaffold: potent design of an artificial cellulosome

DOI： 10.1039/c3cc42614a

Language：Others  

Biosorption of rare earth elements by Escherichia coli
Biosorption, which takes place by passive metal-sequestering, has recently attracted attention as an alternative to conventional separation processes such as precipitation and solvent extraction. In this work, the adsorption behavior of rare earth elements (REEs) onto an Escherichia coli (E. coli) bacterial adsorbent was examined. The adsorption of REEs increased with increasing pH of the feed solution. Also, E. coli showed a higher selectivity for heavier REEs, especially for scandium. To elucidate the adsorption mechanism, the functional groups on the E. coli were chemically modified. By analyzing the adsorption behavior and FT-IR spectra of these E. coli samples before and after adsorption, it was confirmed that REEs are adsorbed at phosphate and carboxyl groups on the surface of E. coli. It is concluded that E. coli is a potential novel adsorbent for REEs. © 2013 The Society of Chemical Engineers, Japan.

DOI： 10.1252/jcej.13we031

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1080/09205063.2012.755600

Language：Others  

Enzymatic preparation of streptavidin-immobilized hydrogel using a phenolated linear poly(ethylene glycol)
Hybrid gels constructed from proteins and polymers have attracted a wide range of attention in the field of biomedicine and bioengineering. We report herein the enzymatic preparation of polymer-protein hybrid hydrogels composed of terminally bis-functionalized linear poly(ethylene glycol) (PEG) and streptavidin (SA). PEG was conjugated with tyramine to introduce terminal phenolic hydroxyl (Ph-OH) groups. A peptide tag containing a tyrosine residue (G4Y-tag) was genetically introduced at the C-terminus of SA. The Ph-OH-modified PEG and G4Y-tagged SA (SA-G4Y) were treated by horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2) to yield (PEG-Ph-OH)-(SA-G4Y) hybrid gels. Biotinylated enhanced green fluorescent protein (biotin-EGFP) was selectively captured in the obtained hybrid gels, indicating that SA-G4Y retained its biological function. The amount of biotin-EGFP immobilized in the hybrid gels depended on the concentration of SA-G4Y. In addition, biotinylated bacterial alkaline phosphatase (biotin-BAP) was immobilized in the hybrid gel. The immobilized biotin-BAP exhibited more than 95% of the initial activity after 5 rounds of recycling. The results suggest the facile functionalization of the hybrid gel with a variety of biotinylated functional molecules. © 2013 Elsevier B.V.

DOI： 10.1016/j.bej.2013.04.007

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c3cc42614a

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2013.04.007

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2013.03.147

Language：Others  

Selective extraction and recovery of rare earth metals from phosphor powders in waste fluorescent lamps using an ionic liquid system
The recycling of rare earth metals from phosphor powders in waste fluorescent lamps by solvent extraction using ionic liquids was studied. Acid leaching of rare earth metals from the waste phosphor powder was examined first. Yttrium (Y) and europium (Eu) dissolved readily in the acid solution; however, the leaching of other rare earth metals required substantial energy input. Ionization of target rare earth metals from the waste phosphor powders into the leach solution was critical for their successful recovery. As a high temperature was required for the complete leaching of all rare earth metals, ionic liquids, for which vapor pressure is negligible, were used as an alternative extracting phase to the conventional organic diluent. An extractant, N, N-dioctyldiglycol amic acid (DODGAA), which was recently developed, showed a high affinity for rare earth metal ions in liquid-liquid extraction although a conventional commercial phosphonic extractant did not. An effective recovery of the rare earth metals, Y, Eu, La and Ce, from the metal impurities, Fe, Al and Zn, was achieved from the acidic leach solution of phosphor powders using an ionic liquid containing DODGAA as novel extractant system. © 2013 Elsevier B.V.

DOI： 10.1016/j.jhazmat.2013.03.026

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.molliq.2013.03.012

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2013.03.147

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.molliq.2013.03.012

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jhazmat.2013.03.026

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2012.06.096

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2012.06.096

Language：Others  

DOI： 10.5360/membrane.38.92

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/bc300606b

Language：Japanese  

Language：Others  

Split Spy0128 as a potent scaffold for protein cross-linking and immobilization
Site-specific cross-linking techniques between proteins and additional functional groups have become increasingly important for expanding the utility of proteins in biochemistry and biotechnology. In order to explore powerful techniques for practical bioconjugation applications, we have validated a technique mediated by a unique property of Streptcoccus pyogenes pilin subunit Spy0128, an autocatalytic intramolecular isopeptide formation in Spy0128. Recently, it has been revealed that Spy0128 can be split into two fragments (split-Spy0128 (residues 18-299 of Spy0128) and isopeptag (residues 293-308 of Spy0128)) that were capable of forming an intermolecular covalent complex. We focused on this unique reconstitution property and first studied the bioconjugation of blue and green fluorescent proteins, enabling the direct monitoring of cross-linking reactions by Förster resonance energy transfer (FRET). A fluorescence lifetime study shows that spatial control of two proteins on the Spy0128 scaffold is possible when one protein is fused to the N-terminus of split-Spy0128 and another one is tethered at the N- or C-terminus of the isopeptag. Furthermore, we demonstrated site-specific protein immobilization mediated by the reconstitution of split-Spy0128 and isopeptag. In this case, a split-Spy0128 mutant with a free N-terminal Cys residue was first immobilized onto beads chemically modified with a maleimide group through a Michael addition process. Then, an isopeptagged protein was successfully immobilized onto the split-Spy0128-immobilized beads. These results suggest that Spy0128 is a potent proteinaceous scaffold available for bioconjugation both in solution and at a solid surface. © 2013 American Chemical Society.

DOI： 10.1021/bc300606b

Language：Others  

Protein supramolecular complex formation by site-specific avidin-biotin interactions
The precise accumulation of protein functions on a nanoscale to fabricate advanced biomaterials has become possible by a bottom-up approach based on molecular self-assembly. The avidin-biotin interaction is widely employed in the design of functional protein self-assemblies. Herein we assessed how the spatial arrangement of the avidin-biotin interaction between protein building blocks affects the formation of a protein supramolecular complex (PSC). The enzymatic site-specific internal labeling of a symmetric protein scaffold, bacterial alkaline phosphatase (AP), with specifically designed biotinylation substrates revealed that the precise positioning of the biotinylation sites on AP and the linker flexibility of the substrate are critical factors for the growth of PSCs in the presence of streptavidin (SA). A potential diagnostic application of the PSCs comprised of AP and SA was demonstrated in an enzyme-linked immunosorbent assay. © 2013 The Royal Society of Chemistry.

DOI： 10.1039/c2ob26625c

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c2ob26625c

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2012.10.002

Language：English   Publishing type：Research paper (scientific journal)  

S/O-nanodispersion electrospun nanofiber mesh effective for sustained release of healthy plasmid DNA with the structuraland functional integrity
Localized and sustained delivery of the therapeutic genes using a solid carrier matrix is a potential approach to develop highly curative treatments. Electrospun nanofiber mesh of biodegradable polymer has been applied extensively as a carrier for localized and sustained delivery of drugs, proteins, and DNA, but it remains difficult to release sufficient amounts of DNA while maintaining structural and functional integrity. To realize the stable sustained release of the healthy plasmid DNA (pDNA) from electrospun fiber mesh, a novel method was examined for loading pDNA into the fibers based on solid-in-oil (S/O) nanodispersion of pDNA in organic solvent for electrospinning polymer solution: S/O nanodispersion electrospinning. A prepared pDNA-loaded fiber mesh made of biodegradable polymer showed sustained release of pDNA without burst release. From luciferase activity-based in vitro transcription-translation assay, pDNA released from meshes of the S/O nanodispersion retained about 60% luciferase activity of control pDNA, whereas pDNA released from the meshes of simple mixing showed only about 5% activity, indicating that S/O nanodispersion electrospinning is effective for loading pDNA into electrospun fiber meshes while maintaining their healthy functions. Effectiveness of S/O nanodispersion electrospinning was verified for fabricating a sustained release carrier matrix for high molecular weight bioactives, including therapeutic genes.

DOI： 10.1080/09205063.2012.755600

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.bej.2012.10.002

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.20.225

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.20.123

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1252/jcej.13we031

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c2md20059g

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c2md20059g

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2012.09.007

Language：English  

DOI： 10.1016/j.ijpharm.2012.09.007

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s10529-012-0998-0

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s10529-012-0998-0

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c2ob25529d

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ejpb.2012.05.016

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ejpb.2012.05.016

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/bc300137s

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/bc300137s

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jconrel.2012.05.001

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.jconrel.2012.05.001

Language：English   Publishing type：Research paper (scientific journal)  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/ac2034198

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/ac2034198

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c2an35172b

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1039/c2cc30618b

Language：English   Publishing type：Research paper (scientific journal)  

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ab.2011.11.028

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.actbio.2011.09.023

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ab.2011.11.028

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.actbio.2011.09.023

Language：Japanese  

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2011.10.003

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.biortech.2011.10.003

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.15261/serdj.19.69

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/chem.201102121

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/chem.201102121

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.ijpharm.2011.08.025

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1021/bc200420v

Language：English   Publishing type：Research paper (scientific journal)