Kyushu University [NOZOMU OKINO (Associate Professor) Faculty of Agriculture, Department of Bioscience and Biotechnology]

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NOZOMU OKINO
Associate Professor
Molecular Bioscience
Department of Bioscience and Biotechnology
Faculty of Agriculture
Last modified date：2024.04.08

Graduate School

Department of Innovative Science and Technology for Bio-industry
Graduate School of Bioresource and Bioenvironmental Sciences

Department of Bioscience and Biotechnology
Graduate School of Bioresource and Bioenvironmental Sciences

Undergraduate School

Department of Bioresource and Bioenvironment
School of Agriculture

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Country of degree conferring institution (Overseas)

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ORCID(Open Researcher and Contributor ID)

Total Priod of education and research career in the foreign country

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Educational

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1.	Ito M, Ishibashi Y, Watanabe T, Iwaki J, Kurita T, Okino N., Assays and Utilization of Enzymes Involved in Glycolipid Metabolism in Bacteria and Fungi, Methods Mol Biol. 2023;2613:229-256, 10.1007/978-1-0716-2910-9_18, 2023.01.
2.	Ito M, Chisada S, Matsunaga N, Okino N. , Vibrio-binding gangliosides in fish intestinal tracts , Glycoconj J. 2023 Jun;40(3):315-322. , 10.1007/s10719-023-10110-1, 2023.06.
3.	Makoto Ito, Nozomu Okino, Motohiro Tani, New insight into the structure, reaction mechanism, and biological functions of neutral ceramidase, Biochimica et Biophysica Acta, 1841, 682–691, 2014.05.
4.	The critical reason for production of ceramidase in Pseudomonas aeruginosa Nozomu Okino.

1.	Nyunoya, Hayato; Ishibashi, Yohei; Ito, Makoto; Okino, Nozomu, Significance of mitochondrial fatty acid β-oxidation for the survivability of Aurantiochytrium limacinum ATCC MYA-1381 during sugar starvation., Bioscience, Biotechnology, and Biochemistry, 10.1093/bbb/zbac141, 86, 11, 1524-1535, 2022.10.
2.	Nyunoya, Hayato; Noda, Tatsuki; Kawamoto, You; Hayashi, Yasuhiro; Ishibashi, Yohei; Ito, Makoto; Okino, Nozomu, Lack of increment 5 Desaturase Activity Impairs EPA and DHA Synthesis in Fish Cells from Red Sea Bream and Japanese Flounder, MARINE BIOTECHNOLOGY, 10.1007/s10126-021-10040-9, 23, 3, 472-481, 2021.06.
3.	Nozomu Okino, Mengbai Li, Qingjun Qu, Tomoko Nakagawa, Yasuhiro Hayashi, Mitsufumi Matsumoto, Yohei Ishibashi, Makoto Ito, Two bacterial glycosphingolipid synthases responsible for the synthesis of glucuronosylceramide and alpha-galactosylceramide, Journal of Biological Chemistry, 10.1074/jbc.RA120.013796, 295, 31, 10709-10725, 2020.07, Bacterial glycosphingolipids such as glucuronosylceramide and galactosylceramide have been identified as ligands for invariant natural killer T cells and play important roles in host defense. However, the glycosphingolipid synthases required for production of these ceramides have not been well-characterized. Here, we report the identification and characterization of glucuronosylceramide synthase (ceramide UDP-glucuronosyltransferase [Cer-GlcAT]) in Zymomonas mobilis, a Gram-negative bacterium whose cellular membranes contain glucuronosylceramide. On comparing the gene sequences that encode the diacylglycerol GlcAT in bacteria and plants, we found a homologous gene that is widely distributed in the order Sphingomonadales in the Z. mobilis genome. We first cloned the gene and expressed it in Escherichia coli, followed by protein purification using nickel-Sepharose affinity and gel filtration chromatography. Using the highly enriched enzyme, we observed that it has high glycosyltransferase activity with UDP-glucuronic acid and ceramide as sugar donor and acceptor substrate, respectively. Cer-GlcAT deletion resulted in a loss of glucuronosylceramide and increased the levels of ceramide phosphoglycerol, which was expressed in WT cells only at very low levels. Furthermore, we found sequences homologous to Cer-GlcAT in Sphingobium yanoikuyae and Bacteroides fragilis, which have been reported to produce glucuronosylceramide and α-galactosylceramide, respectively. We expressed the two homologs of the cer-glcat gene in E. coli and found that each gene encodes Cer-GlcAT and Cer-galactosyltransferase, respectively. These results contribute to the understanding of the roles of bacterial glycosphingolipids in host-bacteria interactions and the function of bacterial glycosphingolipids in bacterial physiology..
4.	Nozomu Okino, Hiroyoshi Wakisaka, Yohei Ishibashi, Makoto Ito, Visualization of Endoplasmic Reticulum and Mitochondria in Aurantiochytrium limacinum by the Expression of EGFP with Cell Organelle-Specific Targeting/Retaining Signals, Marine Biotechnology, 10.1007/s10126-018-9795-7, 1-11, 2018.01.
5.	Nozomu Okino, Makoto Ito, Molecular mechanism for sphingosine-induced Pseudomonas ceramidase expression through the transcriptional regulator SphR, Scientific Reports, 6, 38797, 2016.12.
6.	Inoue T, Okino N, Kakuta Y, Hijikata A, Okano H, Goda HM, Tani M, Sueyoshi N, Kambayashi K, Matsumura H, Kai Y, Ito M., Mechanistic insights into the hydrolysis and synthesis of ceramide by neutral ceramidase., J Biol Chem, 284(14):9566-9577, 2009.04.
7.	N. Okino and M. Ito, Ceramidase Enhances Phospholipase C-induced Hemolysis by Pseudomonas aeruginosa, J. Biol. Chem. , 282, 6021-6030, 2007.03.
8.	H. Monjusho, N. Okino, M. Tani, M. Maeda, M. Yoshida, and M. Ito, A Neutral Ceramidase Homologue of Dictyostelium discoideum Exhibits an Acidic pH Optimum, Biochem. J., 10.1042/BJ20030652, 376, 473-479, 376, pp473-479, 2003.12.
9.	X. He, N. Okino, R. Dhami, A. Dagan, S. Gatt, H. Schulze, K. Sandhoff, and E. H. Schuchman, Purification and Characterization of Recombinant, Human Acid Ceramidase, J. Biol. Chem., 278, pp32978-32986, 2003.08.
10.	N. Okino, X. He, S. Gatt, K. Sandhoff, M. Ito, and E. H. Schuchman, The Reverse Activity of Human Acid Ceramidase, J. Biol. Chem., 10.1074/jbc.M303310200, 278, 32, 29948-29953, 278, pp29948-29953, 2003.08.

1.	Nozomu Okino, Makoto Ito, Identification and functional analysis of the bacterial glucuronosylceramide synthase, 11th Asian Community of Glycoscience and Glycotechnology Conference, 2019.11.
2.	Hayato Nyunoya, Nozomu Okino, Yohei Ishibashi, Makoto Ito, Evaluation of delta5 desaturase activity in marine fish cells, Marine Biotechnology Conference 2019, 2019.09.
3.	Nozomu Okino, Hiroyoshi Wakisaka, Yohei Ishibashi, Makoto Ito, Visualization of endoplasmic reticulum and mitochondria in Aurantiochytrium limacinum ATCC MYA-1381, First International Conference on Labyrinthulean Protists (ICoLP), 2019.08.
4.	Nozomu Okino, Mengbai Li, Yohei Ishibashi, Makoto Ito, Identification and functional characterization of ceramide UDP-glucronosyltransferase in Zymomonas mobilis, 60th International Converence on the Bioscience of Lipids, 2019.06.
5.	Nozomu Okino, Makoto Ito, Identification and functional analysis of diacylglycerol glucuronosyltransferase in Pseudomonas aeruginosa, 2nd Japan-Korea Lipid Joint Symposium, 2018.09.
6.	Nozomu Okino, Makoto Ito, Transcriptional regulation of Pseudomonas ceramidase through the sphingosine-specific transcriptional regulator SphR , 58th International Conference on the Bioscience of Lipids, 2017.09.
7.	沖野望, Yoshimitsu Kakuta, 鵜木(加藤)陽子, 大坪　怜央, 石橋洋平, 小林　宇太郎, makoto kimura, 伊東信, Crystal structures of unique glycosphingolipid-degrading enzymes, The 7thACGG conference, 2015.11.
8.	沖野望, 伊東信, Marine biotechnology: New understanding and developments, PKNU – KU Joint Symposium SMART Oceans and Fisheries, 2015.10.
9.	沖野望, 伊東信, Identification of a transcriptional regulator SphR for ceramidase expression in Pseudomonas aeruginosa, 第10回スフィンゴテラピィ研究会, 2015.06.