Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Yuji Tsutsumi Last modified date:2020.06.17

Professor / Sustainable Bioresources Science / Department of Agro-environmental Sciences / Faculty of Agriculture


Papers
1. Manami Takeuchi, Watanabe Atsushi, Miho Tamura, Yuji Tsutsumi, The gene expression analysis of Arabidopsis thaliana ABC transporters by real-time PCR for screening monolignol-transporter candidates, Journal of Wood Science, 10.1007/s10086-018-1733-9, 64, 477-484, 2018.10, The transport of monolignols from the cytosol to the cell wall is essential for lignin synthesis. The ATP-binding cassette (ABC) transporters may be involved in the transport of lignin precursors. ABC transporter genes subjected to expression analysis were chosen based on two criteria for screening candidate transporter genes related to lignification. The expression levels of 15 target genes in five plant organs were analyzed by real-time PCR. Five transporter genes (ABCG29, ABCG30, ABCG33, ABCG34, and ABCG37), which were simultaneously expressed with the reference genes, were selected as candi- dates. The candidate gene expression levels in root tissues of T-DNA insertion mutants were determined by semi-quantitative reverse transcription PCR. ABCG30 was more highly expressed in the abcg34 mutant than in the wild-type plants, while the expression of ABCG34 was twofold higher in the abcg30 mutant plants than in the wild-type plants. Thus, the expression of ABCG30 and ABCG34 may affect each other. There was no significant change in lignin content and composition in the single-gene knockout mutants of the candidate transporter genes, which suggested that each candidate gene did not solely contribute to lignin synthesis..
2. Jun Shigeto, Hiroki Honjo, Koki Fujita and Yuji Tsutsumi, Generation of lignin polymer models via dehydrogenative polymerization of coniferyl alcohol and syringyl alcohol via several plant peroxidases involved in lignification and analysis of the resulting DHPs by MALDI-TOF analysis, Holzforschung, 10.1515/hf-2017-0125, 72, 4, 267-274, 2017.12, The mechanism of lignin dehydrogenative poly- merization (DHP), made by means of horseradish peroxi- dase (HRP), was studied in comparison with other plant peroxidases. Interestingly, HRP is efficient for guaiacyl type polymer formation (G-DHPs), but is not efficient in the case of syringyl type DHPs (S-DHPs). It was previously demonstrated that lignification-related Arabidopsis thali- ana peroxidases, AtPrx2, AtPrx25 and AtPrx71, and cati- onic cell-wall-bound peroxidase (CWPO-C) from Populus alba are successful to oxidize syringyl- and guaiacyl-type monomers and larger lignin-like molecules. This is the reason why in the present study the DHP formation by means of these recombinant peroxidases was tested, and all these enzymes were successful for formation of both G-DHP and S-DHP in acceptable yields. CWPO-C led to S-DHP molecular size distribution similar to that of iso- lated lignins..
3. Manami Takeuchi, Takahiro Kegasa, Watanabe Atsushi, Miho Tamura, Yuji Tsutsumi, Expression analysis of transporter genes for screening candidate monolignol transporters using Arabidopsis thaliana cell suspensions during tracheary element differentiation, Journal of Plant Research, 10.1007/s10265-017-0979-4, 131, 297-305, 2018.03, The mechanism of monolignol transportation from the cytosol to the apoplast is still unclear despite being an essential step of ligni cation. Recently, ATP-binding cas- sette (ABC) transporters were suggested to be involved in monolignol transport. However, there are no reliable clues to the transporters of the major lignin monomers coniferyl and synapyl alcohol. In this study, the ligni cation progress of Arabidopsis cultured cells during tracheary element dif- ferentiation was monitored. The expression of selected trans- porter genes, as well as ligni cation and cell-wall formation related genes as references, in di erentiating cultured cell samples harvested at 2-day intervals was analyzed by real- time PCR and the data were statistically processed. The cell wall formation transcription factor MYB46, programmed- cell death related gene XCP1 and lignin polymerization peroxidase AtPrx25 were classi ed into the same cluster. Furthermore, the cluster closest to the abovementioned cluster contained the lignin synthesis transcription factor..
4. Jun Shigeto, Yukie Ueda, Shinya Sasaki, Koki Fujita, Yuji Tsutsumi, Enzymatic activities for lignin monomer intermediates highlight the biosynthetic pathway of syringyl monomers in Robinia pseudoacacia, Journal of Plant Research, 10.1007/s10265-016-0882-4, 130, 1, 203-210, 2017.01, Most of the known 4-coumarate:coenzyme A ligase (4CL) isoforms lack CoA-ligation activity for sinapic acid. Therefore, there is some doubt as to whether sinapic acid contributes to sinapyl alcohol biosynthesis. In this study, we characterized the enzyme activity of a protein mixture extracted from the developing xylem of Robinia pseudoacacia. The crude protein mixture contained at least two 4CLs with sinapic acid 4-CoA ligation activity. The crude enzyme preparation displayed negligible sinapaldehyde dehydrogenase activity, but showed ferulic acid 5-hydroxylation activity and 5-hydroxyferulic acid O-methyltransferase activity; these activities were retained in the presence of competitive substrates (coniferaldehyde and 5-hydroxyconiferaldehyde, respectively). 5-Hydroxyferulic acid and sinapic acid accumulated in the developing xylem of R. pseudoacacia, suggesting, in part at least, sinapic acid is a sinapyl alcohol precursor in this species..
5. Premjet Siripong, Premjet Duangporn, Eri Takata, Yuji Tsutsumi, Phosphoric acid pretreatment of Achyranthes aspera and Sida acuta weed biomass to improve enzymatic hydrolysis, Bioresource Technology, 10.1016/j.biortech.2015.12.037, 203, 303-308, 2016.03, Achyranthes aspera and Sida acuta, two types of weed biomass are abundant and waste in Thailand. We focus on them as novel feedstock for bio-ethanol production because they contain high-cellulose content (45.9% and 46.9%, respectively) and unutilized material. Phosphoric acid (70%, 75%, and 80%) was employed for the pretreatment to improve by enzymatic hydrolysis. The pretreatment process removed most of the xylan and a part of the lignin from the weeds, while most of the glucan remained. The cellulose conversion to glucose was greater for pretreated A. aspera (86.2 ± 0.3%) than that of the pretreated S. acuta (82.2 ± 1.1%). Thus, the removal of hemicellulose significantly affected the efficiency of the enzymatic hydrolysis. The scanning electron microscopy images showed the exposed fibrous cellulose on the cell wall surface, and this substantial change of the surface structure contributed to improving the enzyme accessibility..
6. Yuichi Murayama, Miyako Yoshioka, Hiroyuki Okada, Eri Takata, Kentaro Masujin, Yoshifumi Iwamaru, Noriko Shimozaki, Tomoaki Yamamura, Takashi Yokoyama, Shirou Mohri, Yuji Tsutsumi, Subcritical Water Hydrolysis Effectively Reduces the In Vitro Seeding Activity of PrPSc but Fails to Inactivate the Infectivity of Bovine Spongiform Encephalopathy Prions, PLoS One, 10.1371/journal.pone.0144761, 10, 12, e0144761, 2015.12, The global outbreak of bovine spongiform encephalopathy (BSE) has been attributed to the recycling of contaminated meat and bone meals (MBMs) as feed supplements. The use of MBMs has been prohibited in many countries; however, the development of a method for inactivating BSE prions could enable the efficient and safe use of these products as an organic resource. Subcritical water (SCW), which is water heated under pressure to maintain a liquid state at temperatures below the critical temperature (374°C), exhibits strong hydrolytic activity against organic compounds. In this study, we examined the residual in vitro seeding activity of protease-resistant prion protein (PrPSc) and the infectivity of BSE prions after SCW treatments. Spinal cord homogenates prepared from BSE-infected cows were treated with SCW at 230–280°C for 5–7.5 min and used to intracerebrally inoculate transgenic mice overexpressing bovine prion protein. Serial protein misfolding cyclic amplification (sPMCA) analysis detected no PrPSc in the SCW-treated homogenates, and the mice treated with these samples survived for more than 700 days without any signs of disease. However, sPMCA analyses detected PrPSc accumulation in the brains of all inoculated mice. Furthermore, secondary passage mice, which inoculated with brain homogenates derived from a western blotting (WB)-positive primary passage mouse, died after an average of 240 days, similar to mice inoculated with untreated BSE-infected spinal cord homogenates. The PrPSc accumulation and vacuolation typically observed in the brains of BSE-infected mice were confirmed in these secondary passage mice, suggesting that the BSE prions maintained their infectivity after SCW treatment. One late-onset case, as well as asymptomatic but sPMCA-positive cases, were also recognized in secondary passage mice inoculated with brain homogenates from WB-negative but sPMCA-positive primary passage mice. These results indicated that SCW-mediated hydrolysis was insufficient to eliminate the infectivity of BSE prions under the conditions tested..
7. Jun Shigeto, Itoh Yoshitaka, Hrao Sakie, Ohhira Kaori, Koki Fujita, Yuji Tsutsumi, Simultaneously disrupting AtPrx2, AtPrx25 and AtPrx71 alters lignin content and structure in Arabidopsis stem, JOURNAL OF INTEGRATIVE PLANT BIOLOGY, 10.1111/jipb.12334, 57, 4, 349-356, 2015.04, Lignins are aromatic heteropolymers that arise from oxidative coupling of lignin precursors, including lignin monomers (p-coumaryl, coniferyl, and sinapyl alcohols), oligomers, and polymers. Whereas plant peroxidases have been shown to catalyze oxidative coupling of monolignols, the oxidation activity of well-studied plant peroxidases, such as horseradish peroxidase C (HRP-C) and AtPrx53, are quite low for sinapyl alcohol. This characteristic difference has led to controversy regarding the oxidation mechanism of sinapyl alcohol and lignin oligomers and polymers by plant peroxidases. The present study explored the oxidation activities of three plant peroxidases, AtPrx2, AtPrx25, and AtPrx71, which have been already shown to be involved in lignification in the Arabidopsis stem. Recombinant proteins of these peroxidases (rAtPrxs) were produced in Escherichia coli as inclusion bodies and successfully refolded to yield their active forms. rAtPrx2, rAtPrx25, and rAtPrx71 were found to oxidize two syringyl compounds (2,6-dimethoxyphenol and syringaldazine), which were employed here as model monolignol compounds, with higher specific activities than HRP-C and rAtPrx53. Interestingly, rAtPrx2 and rAtPrx71 oxidized syringyl compounds more efficiently than guaiacol. Moreover, assays with ferrocytochrome c as a substrate showed that AtPrx2, AtPrx25, and AtPrx71 possessed the ability to oxidize large molecules. This characteristic may originate in a protein radical. These results suggest that the plant peroxidases responsible for lignin polymerization are able to directly oxidize all lignin precursors..
8. Takako Harada, Koki Fujita, Jun Shigeto, Yuji Tsutsumi, Stereo-selective oxidations of terpinolene by cytochrome P450 monooxygenases in the microsomal fraction of Cupressus lusitanica cultured cells, Journal of Wood Science, 10.1371/journal.pone.0105332, 9, 8, e105332- e105332, 2014.08.
9. Jun Shigeto, Mariko Nagano, Koki Fujita, Yuji Tsutsumi, Catalytic Profile of Arabidopsis Peroxidases, AtPrx-2, 25 and 71, Contributing to Stem Lignification , PLoS ONE, 10.1371/journal.pone.0105332, 9, 8, e105332, 2014.08, Lignins are aromatic heteropolymers that arise from oxidative coupling of lignin precursors, including lignin monomers (p-coumaryl, coniferyl, and sinapyl alcohols), oligomers, and polymers. Whereas plant peroxidases have been shown to catalyze oxidative coupling of monolignols, the oxidation activity of well-studied plant peroxidases, such as horseradish peroxidase C (HRP-C) and AtPrx53, are quite low for sinapyl alcohol. This characteristic difference has led to controversy regarding the oxidation mechanism of sinapyl alcohol and lignin oligomers and polymers by plant peroxidases. The present study explored the oxidation activities of three plant peroxidases, AtPrx2, AtPrx25, and AtPrx71, which have been already shown to be involved in lignification in the Arabidopsis stem. Recombinant proteins of these peroxidases (rAtPrxs) were produced in Escherichia coli as inclusion bodies and successfully refolded to yield their active forms. rAtPrx2, rAtPrx25, and rAtPrx71 were found to oxidize two syringyl compounds (2,6-dimethoxyphenol and syringaldazine), which were employed here as model monolignol compounds, with higher specific activities than HRP-C and rAtPrx53. Interestingly, rAtPrx2 and rAtPrx71 oxidized syringyl compounds more efficiently than guaiacol. Moreover, assays with ferrocytochrome c as a substrate showed that AtPrx2, AtPrx25, and AtPrx71 possessed the ability to oxidize large molecules. This characteristic may originate in a protein radical. These results suggest that the plant peroxidases responsible for lignin polymerization are able to directly oxidize all lignin precursors..
10. Eri Takata, Tatsushi Tsuruoka, Ken Tsutsumi, Yuji Tsutsumi, Kenji Tabata, Production of xylitol and tetrahydrofurfuryl alcohol from xylan in napier grass by a hydrothermal process with phosphorus oxoacids followed by aqueous phase hydrogenation, Bioresource Technology, 10.1016/j.biortech.2014.05.112, 167, 74-80, 2014.06, The production of xylitol and tetrahydrofurfuryl alcohol (THFA) from napier grass was studied using two
steps: a hydrothermal process with phosphorus oxoacids followed by aqueous phase hydrogenation with
Pd/C. Xylose obtained from the napier grass by the hydrothermal treatment with 3.0 wt% phosphorous
acid was subsequently converted into xylitol at 51.6% yield of the xylan in napier grass by hydrogenation
with 5.0 wt% Pd/C. The furfural produced from napier grass with a 3.0 wt% phosphoric acid treatment was
also directly subjected to the hydrogenation as a hydrolysate to yield 41.4% THFA based on the xylan in
napier grass. The yields of xylitol and THFA obtained by hydrogenation using the napier grass hydrolysate
containing xylose or furfural were almost the same as those of hydrogenation using commercial materi-
als. To our knowledge, this is the first report on the production of THFA in high yield by hydrogenation
directly from biomass hydrolysate..
11. Koki Fujita, Yasufumi Bunyu, Ken-ichi Kuroda, Tatsuya Ashitani, Jun Shigeto, Yuji Tsutsumi, A novel synthetic pathway for tropolone ring formation via the olefin monoterpene intermediate terpinolene in cultured Cupressus lusitanica cells, Journal of Plant Physiology, 10.1016/j.jplph.2013.12.016, 171, 8, 610-614, 2014.03.
12. Eri Takata, Ken Tsutsumi, Yuji Tsutsumi, Kenji Tabata, Production of monosaccharides from napier grass by hydrothermal process with phosphoric acid, Bioresource Technology, 10.1016/j.biortech.2013.05.112, 143, 53-58, 2013.09.
13. Zhang Fujia, Junko Kajiwara, Mineko Ohira, Yasuhiro Mori, Yuji Tsutsumi, RYUICHIRO KONDO, Metabolites from Resistant and Susceptible Pinus thunbergii after Inoculation with Pine Wood Nematode, American Journal of Plant Science, 10.4236/ajps.2013.43065, 4, 3, 512-518, 2013.03.
14. Zhang Fujia, Yasuhiro Mori, Yuji Tsutsumi, RYUICHIRO KONDO, A rapid in vitro bioassay system for testing resistance factors of pine trees to Bursaphelenchus xylophilus, Nematology, 10.1163/15685411-00002709, 15, 6, 665-670, 2013.06.
15. Jun Shigeto, Yuko Kiyonaga, Koki Fujita, RYUICHIRO KONDO, Yuji Tsutsumi, Putative Cationic Cell-Wall-Bound Peroxidase Homologues in Arabidopsis, AtPrx2, AtPrx25, and AtPrx71, Are Involved in Lignification, J. Agric. Food Chem., 10.1021/jf400426g, 61, 16, 3781-3788, 2013.04, The final step of lignin biosynthesis, which is catalyzed by a plant peroxidase, is the oxidative coupling of the
monolignols to growing lignin polymers. Cationic cell-wall-bound peroxidase (CWPO-C) from poplar callus is a unique enzyme
that has oxidative activity for both monolignols and synthetic lignin polymers. This study shows that putative CWPO-C
homologues in Arabidopsis, AtPrx2, AtPrx25, and AtPrx71, are involved in lignin biosynthesis. Analysis of stem tissue using the
acetyl bromide method and derivatization followed by the reductive cleavage method revealed a significant decrease in the total
lignin content of ATPRX2 and ATPRX25 deficient mutants and altered lignin structures in ATPRX2, ATPRX25, and ATPRX71
deficient mutants. Among Arabidopsis peroxidases, AtPrx2 and AtPrx25 conserve a tyrosine residue on the protein surface, and
this tyrosine may act as a substrate oxidation site as in the case of CWPO-C. AtPrx71 has the highest amino acid identity with
CWPO-C. The results suggest a role for CWPO-C and CWPO-C-like peroxidases in the lignification of vascular plant cell walls..
16. Shigeto Jun;Itoh Yoshitaka;Tsutsumi Yuji;et al., Identification of Tyr74 and Tyr177 as substrate oxidation sites in cationic cell wall-bound peroxidase from Populus alba L., FEBS JOURNAL, 10.1111/j.1742-4658.2011.08429x, 279, 2, 348-357, 2012.01.
17. Y. Mori , F. Miyahara , Y. Tsutsumi and R. Kondo , Effects of combinational treatment with ethephon and indole-3-butyric acid on adventitious rooting of Pinus thunbergii cuttings
, Plant Growth Regulation, 63(3) 271-278, 2011.03.
18. Y. Mori, F. Miyahara, Y. Tsutsumi, R. Kondo , Relationship between resistance to pine wilt disease and the migration or proliferation of pine wood nematodes.
, European Journal of Plant Pathology, 122(4), 529-538 , 2008.04.
19. I. Kamei, C. Daikoku, Y. Tsutsumi, R. Kondo , Saline-dependent regulation of manganese peroxidase genes in the hypersaline-tolerant white rot fungus Phlebia sp. MG-60, Applied and Environmental Microbiology, 74(9), 2709-2716 , 2008.04.
20. S. Sasaki, D. Nonaka, H. Wariishi, Y. Tsutsumi, R. Kondo , Role of Tyr residues on the protein surface of cationic cell-wall-peroxidase(CWPO-C) from poplar; Potentially unique oxidation sites for oxidative polymerization of lignin.
, Phytochemistry, 10(4), 651-8, 2008.04.
21. J. Kim, Y. Kim, Y. J. Lee, M. Kobayashi, Y. Tsutsumi, R. Kondo, S. K. Lee and K.-S. Soh., Spontaneous Ultraweak Photon Emission during the Growth of the Cell Population of Cultured HeLa Cell Line, J. Health Sci. , 53, 481-485., 2007.04.
22. Sasaki S, Shimizu S, Wariishi H, Tsutsumi Y, Kondo R, Transcriptional and translational analyses of poplar anionic peroxidase isoenzymes. J. Wood. Sci., , J. Wood. Sci., 53:427-435, 2007.12.
23. Y. Kim, J. Han, S. S. Lee, K. Shimizu, Y. Tsutsumi, R. Kondo, Steroid 9a-hydroxylation during testosterone degradation by resting Rhodococcus equi Cells. , Arch. Pharm. Chem. Life Sci., 340, 209-214 (2007), 2007.10.
24. Shinya Sasaki, Daisuke Nonaka, Hiroyuki Wariishi, Yuji Tsutsumi, Ryuichiro Kondo, Role of Tyr residues on the protein surface of cationic cell-wall-peroxidase (CWPO-C) from poplar: Potentially unique oxidation sites for oxidative polymerization of lignin., Phytochemistry, 69 (2) 348-355 (2008), 2007.03.
25. K. Oshiman, Y. Tsutsum, T. Nishida, Y. Matsumura, Isolation and characterization of a novel bacterium, Sphingomonas bisphenolicum strain AO1, that degrades bisphenol A , Biodegradation, 18, 247-255 , 2007.02.
26. S. Sasaki, K. Baba, T. Nishida, Y. Tsutsumi, R. Kondo, The cationic cell-wall-peroxidase having oxidation ability for polymeric substrate participates in the late stage of lignification of Populus alba L, Plant Mol. Biol., 62(6), 797-807, 2006.12.
27. K. Hamada, T. Nishida, K. Yamauchi, K. Fukushima, R. Kondo, Y. Tsutsumi, 4-Coumarate:coenzyme A ligase in black locust (Robinia pseudoacacia) catalyses the conversion of sinapate to sinapoyl-CoA., Journal of Plant Research, 10.1007/s10265-004-0159-1, 117, 4, 303-310, 117(4) 303-310 (2004), 2004.08.
28. S. Sasaki, T. Nishida, Y. Tsutsumi, R. Kondo., Lignin dehydrogenative polymerization mechanism: a poplar cell wall peroxidase directly oxidizes polymer lignin and produces in vitro dehydrogenative polymer rich in beta-O-4 linkage., FEBS Letters, 10.1016/S0014-5793(04)00224-8, 562, 1-3, 197-201, 562: 197-201 (2004), 2004.04.
29. K Hamada, Y. Tsutsumi, T. Nishida, Treatment of poplar callus with ferulic and sinapic acids II: Effect on related monolignol biosynthetic pathway enzyme activities., J. Wood Sci., 10.1007/s10086-002-0475-9, 49, 4, 366-370, 49:366-370 (2003)
 , 2003.08.
30. K Hamada, Y. Tsutsumi, K. Yamauchi, K. Fukushima, T. Nishida, Treatment of poplar callus with ferulic and sinapic acids I: Incorporation and enhancement of lignin biosynthesis., J. Wood Sci., 10.1007/s10086-002-0477-7, 49, 4, 333-338, 49:333-338 (2003)
 , 2003.08.
31. K.Yamauchi, S. Yasuda, K. Hamada, Y. Tsutsumi, K. Fukushima, Multiform biosynthetic pathway of syringyl lignin in angiosperms, Planta, 10.1007/s00425-002-0865-7, 216, 3, 496-501, 216(3):496-501 (2003), 2003.01.
32. W. Aoyama, S. Sasaki, S. Matsumura, T. Mitsunaga, H. Hirai, Y. Tsutsumi, T. Nishida, Sinapyl alcohol-specific peroxidase isoenzyme catalyzes the formation of the dehydrogenative polymer from sinapyl alcohol., J. Wood Sci., 10.1007/BF00766646, 48, 6, 497-504, 48(6):497-504 (2002), 2002.12.
33. Aoyama W., Matsumura A., Tsutsumi Y., Nishida T, Lignification and peroxidase in tension wood of Eucalyptus viminalis seedlings., J. Wood Sci., 47:419-424 (2001), 2001.01.
34. Tsutsumi Y., Haneda T., Nishida T., Removal of estrogenic activities of bisphenol a and nonylphenol by oxidative enzymes from lignin-degrading basidiomycetes., Chemosphere, 42(3): 271-276, (2001), 2001.01.