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Makoto Ito Last modified date:2018.06.15

Graduate School
Undergraduate School

Academic Degree
Ph. D.
Country of degree conferring institution (Overseas)
Field of Specialization
Functional Biochemistry, Glycobiology/Lipid Biology, Marine Resource Chemistry,
Total Priod of education and research career in the foreign country
Outline Activities
Dr. Makoto Ito, Professor of Molecular Bioscience, Department of Bioscience and Biotechnology, Kyushu University, is involved in the research focussing on the metabolism and functions of sphingolipids and glycolipids. Recently, his research group disclosed the new system for quality control of glucosylceramide in Cryptococcus neoformans, in which a novel enzyme EGCrP was discovered (JBC, 2011). Furthermore, the metabolic pathway of sterylglucoside in fungi was uncovered by his group (JBC, 2015). These findings are highly expected to lead the develop of new drugs for pathogenic fungi. Recently, his research interest expanded to the metabolism of phospholipids, glycerolipids and fatty acids in marine thraustochytrids to supply the new source for energy and medicine/nutrition. For education, he teaches the biochemistry and marine chemistry for graduate and undergraduate students. He was reserved as an council for Japanese Society for Biochemistry, Japanese Carbohydrate Society (JCS) and Japanese Society for Lipid Biochemistry (JSLB) and he chaired the domestic meeting for JCS in 2007 and JSLB in 2012. He was also appointed as an associate editor for Journal of Biochemistry, Glycoconjugate Journal and a corresponding member for International Conference of Lipid Bioscience.
Research Interests
  • Studies on the Metabolic Pathway of Glycolipids and its Biological Significance in Pathogenic Fungi
    keyword : glycolipids, metabolism, vacuole formation, anti-biotics
  • Study on the Production of Bioactive Lipids Using Marine Traustochytrids.
    keyword : Thraustochytrids, Biosynthesis, Metabolism, Lipids, n-3PUFA, DHA, EPA, Phospholipids, Triacylglycerol, Sphingolipids, Lipid Droplet
  • Study on Synthesis of Polyunsaturated Fatty Acids in Marine Microbes.
    keyword : Thraustochytrids, n-3PUFA, EPA, PUFA-synthase, Elongase/Desaturase Pathway
  • Studies on structure and functions of glycolipid-metabolizing enzymes in fungi.
    keyword : glycosphingolipids, metabolism, infection diseases, cryptococcosis, endoglycoceramidase, EGCrP, glucosylceramide, sterylgucoside, vacuole
  • Structures, functions and applications of sphingolipid-signaling enzymes.
    keyword : sphingolipids, glycolipids, ceramide, cellular signaling, metabolic enzymes, microdomain, inhibitors, zebrafish, embryogenesis
    1997.04Molecular Evolution, Functions and Applications of Sphingolipid-signaling Enzymes..
  • Study on O-glycosylation of glycoconjugates.
    keyword : O-glycans, O-glycanse, mucine, glycosphingolipids, cellular traffic, microdomain, neutral ceramidase, microbe adhesion, fish pathogens
    2001.01Structures and Functions of Ser/Thr- or Ceramide-linked O-Glycans..
  • Study on the interaction and adhesion of microbes to glycosphingolipids.
    keyword : glycolipids, ganglioside, lectin, marine lactobacillus, pro-biotics
  • Study on development and applications of marine genomes.
    keyword : genome, microbes, invertebrates, glycolipids/sphingolipids, seaweeds, enzymes, lectins
    2002.04Reserch and Development of Marine Genomes and Genes..
  • Functional analysis of sphingolipid-signaling enzymes using early development of zebrafish.
    keyword : early development, anti-morphorino oligo, gene expression, signal transduction, ceramide, ceramidase, glycolipid synthesis, glucosylceramide
    2000.01Functional Analyses of Genes Encoding Sphingolipid-signaling Enzymes using Zebrafish Early Development and Slime Mold Morphogenesis..
Current and Past Project
  • Systematic production of HUFA in thraustochytrids based on genome data and gene manipulaton.
  • Sphingolipid Biology: Membrane Domain Dynamics and Multifunctional Signaling.
Academic Activities
1. Makoto Ito, Degrading of Glycolipids, Elsevier, volume 3, 193-208, 2007.10.
2. Makoto Ito, Motohiro Tani and Yukihiro Yoshimura, Sphingolipid Biology-
Neutral ceramidase as an integral modulator for the generation of S!P and S1P-mediated signaling
, Springer, pp183-196, 2006.03.
3. Makoto Ito, Nozomu Okino, Motohiro Tani, Susumu Mitsutake, Katsuhiro Kita, Ceramide Signaling -
Molecular evolution of neutral ceramidase: From bacteria to mammals
, Landes Bioscience, 2002.06.
1. A novel metabolic pathway of glucosylceramide that involves Klotho-related protein KLrP.
2. Catalytic mechanism, topology and biological functions of neutral ceramidase uncovered by X-ray crystal structure.
3. Recruitment of the neutral ceramidase to plasma membranes as a type II membrane protein by O-glycosylation of mucin-like domain.
4. Molecular evolution of neutral ceramidase: Signaling molecule and viruence factor.
1. Watanabe T, Sakiyama R, Iimi Y, Sekine S, Abe E, Nomura H, Ishibashi Y, Okino N, Ito M, Regulation of TG accumulation and lipid droplet morphology by the novel TLDP1 in Aurantiochytrium limacinum F26-b, Journal Lipid Research, 2017.10.
2. Nozomu Okino, Makoto Ito, Molecular mechanism for sphingosine-induced Pseudomonas ceramidase expression through the transcriptional regulator SphR, 10.1038/screp38797, 2016.11.
3. Takashi Watanabe, Motohiro Tani, Yohei Ishibashi, Ikumi Endo, NOZOMU OKINO, Makoto Ito, Ergosteryl-β-glucosidase (Egh1) involved in sterylglucoside catabolism and vacuole formation in Saccharomyces cerevisiae. 
, Glycobiology, 25, 10, 1079-1089, 2015.04.
4. Takashi Watanabe, Tomohiro Ito, Hatsumi Goda, Yohei Ishibashi, NOZOMU OKINO, Makoto Ito, Sterylglucoside catabolism in Cryptococcus neoformans with endoglycocoramidase-related protein2(EGCrP2), the first steryl-β-glucosidase identifed in fungi. 
, Journal of Biological Chemistry, 290, 1005-1019, 2015.01, Cryptococcosis is an infectious disease caused by pathogenic fungi such as Cryptococcus neoformans and C. gattii. The ceramide structure (methyl-d18:2/h18:0) of C. neoformans glucosylceramide (GlcCer) is characteristic and strongly related to their pathogenicity. Recently, we reported that endoglycoceramidase-related protein 1 (EGCrP1) is a glucocerebrosidase in C. neoformans and involved in the quality control of GlcCer by eliminating immature GlcCer during the synthesis of GlcCer (Ishibashi et al, J. Biol. Chem., 2012). We report here the identification and characterization of EGCrP2, a homologue of EGCrP1, as an enzyme responsible for sterylglucoside catabolism in C. neoformans. In contrast to EGCrP1 specific to GlcCer, EGCrP2 was found to hydrolyze a variety of β-glucosides including cholesteryl-β-glucoside, ergosteryl-β-glucoside, sitosteryl-β-glucoside, GlcCer, and para-nitrophenyl-β-glucoside, but not α-glucosides or β-galactosides, under the acidic condition. Disruption of the EGCrP2 gene (egcrp2) resulted in the accumulation of glycolipid, and the structure was determined following purification to ergosteryl-3-β-glucoside by mass spectrometric and two-dimensional nuclear magnetic resonance analyses. This glycolipid was found to accumulate in vacuole where EGCrP2 is localized. These results indicated that EGCrP2 is involved in the catabolism of ergosteryl-β-glucoside in the vacuole of C. neoformans. The egcrp2-disrupted mutants showed distinct growth arrest, dysfunction of cell budding, and an abnormal vacuole morphology, suggesting that EGCrP2 is a promising target for anti-cryptococcal drugs. EGCrP2, classified into glycohydrolase family 5, is the first identified steryl-β-glucosidase and a missing link in steryl glucoside metabolism in fungi. .
5. Yohei Ishibashi, NOZOMU OKINO, Makoto Ito, A novel ether-linked phytol-containing digalactosylglycerolipid in the marine green alga, Ulva pertusa., Biochem Biophys Res Commun, 10.1016/j.bbrc.2014.08.056., 452, 873-80, 2014.10.
6. Eriko Abe, Makoto Ito, Novel Lysophospholipid Acyltransferase PLAT1 of Aurantiochytrium limacinum F26-b Responsible for Generation of Palmitate-Docosahexaenoate-Phosphatidylcholine and Phosphatidylethanolamine, PLOS ONE, 10.1371/journal.pone.0102377, 9, 8, 2014.08.
7. Makoto Ito, NOZOMU OKINO, Motohiro Tani, New insights into the structure, reaction mechanism, and biological functions of neutral ceramidase, Biochimica ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 1841, 5, 682-691, 2014.05.
8. Junichiro Ohara, Keishi Sakaguchi, NOZOMU OKINO, Makoto Ito, Two fatty acid elongates possessing C18-Δ6/C18-Δ9/C20-Δ5 or C16-Δ9 Elongase activity in Thraustochytrium sp. ATCC 26185, Marine Biotechnology, 10.1007/s10126-013-9496-1, 2013.04.
9. Chisada, Shin-ichi, Shimizu, Kohei, Kamada, Haruna, Matsunaga, Naoyuk, NOZOMU OKINO, Makoto Ito, Vibrios adhere to epithelial cells in the intestinal tract of red sea bream, Pagrus major, utilizing GM4 as an attachment site, FEMS MICROBIOLOGY LETTERS, 10.1111/1574-6968.12082, 341, 1, 18-26, 2013.04.
10. Ishibashi, Yohei, Ikeda, Kazutaka, Sakaguchi, Keishi, NOZOMU OKINO, Taguchi, Ryo, Makoto Ito, Quality Control of Fungus-specific Glucosylceramide in Cryptococcus neoformans by Endoglycoceramidase-related Protein 1 (EGCrP1), JOURNAL OF BIOLOGICAL CHEMISTRY, 10.1074/jbc.M111.311340, 287, 1, 368-381, 2012.01.
11. Sakaguchi, Keishi, Matsuda, Takanori;, Kobayashi, Takumi, Ohara, Junichiro, Hamaguchi, Rie, Abe, Eriko, Hayashi, Masahiro, Honda, Daiske, NOZOMU OKINO, Makoto Ito, Versatile Transformation System That Is Applicable to both Multiple Transgene Expression and Gene Targeting for Thraustochytrids, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 10.1128/AEM.07129-11, 78, 9, 3193-3202, 2012.05.
12. Takanori Matsuda, Hamaguchi, Rie, Kobayashi, Takumi, Abe, Eriko, Hama, Yoichiro, Hayashi, Masahiro, Honda, Daiske, NOZOMU OKINO, Makoto Ito, Analysis of Delta 12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304, JOURNAL OF LIPID RESEARCH, 10.1194/jlr.M024935, 53, 6, 1210-1222, 2012.06.
13. Ishibashi, Yohei;, Kobayashi, Utaro, Hijikata, Atsushi, Sakaguchi, Keish, 合田 初美, NOZOMU OKINO, Makoto Ito, Preparation and characterization of EGCase I, applicable to the comprehensive analysis of GSLs, using a rhodococcal expression system, JOURNAL OF LIPID RESEARCH, 10.1194/jlr.D028951, 53, 10, 2242-2251, 2012.10.
14. Kobayashi T, Sakaguchi K, Matsuda T, Abe E, Hama Y, Hayashi M, Honda D, Okita Y, Sugimoto S, Okino N, Ito M.
, Increase of EPA in Thraustochytrids through Expression of a Fatty Acid Δ5 Desaturase Gene Driven by the Thraustochytrid Ubiquitin Promoter.
, Appl. Environ. Microbiol. , 77, 11, 3870-3876, 2011.11.
15. Sumida T, Fujimoto K, Ito M, Molecular Cloning and Catalytic Mechanism of a Novel Glycosphingolipid-degrading β-N-Acetylgalactosaminidase from Paenibacillus sp. TS12.
J. Biol. Chem. , 286, 16, 14065-14072, 2011.04.
16. Takara T, Nakagawa T, Isobe M, Okino N, Ichinose S, Omori A, Ito M., Purification, molecular cloning, and application of a novel sphingomyelin-binding protein (clamlysin) from the brackishwater clam, Corbicula japonica.
, Biochim. Biophys. Acta. , 1811, 5, 323-332, 2011.05.
17. Togayachi A, Kozono Y, Ikehara Y, Ito H, Suzuki N, Tsunoda Y, Abe S, Sato T, Nakamura K, Suzuki M, Goda HM, Ito M, Kudo T, Takahashi S, Narimatsu H.
, Lack of lacto/neolacto-glycolipids enhances the formation of glycolipid-enriched microdomains, facilitating B cell activation.
, Proc. Natl. Acad. Sci. USA, 107, 26, 11900-11905 , 2010.04, In a previous study, we demonstrated that β1,3-N-acetylglucosaminyltransferase
5 (B3gnt5) is a lactotriaosylceramide (Lc3Cer) synthase
that synthesizes a precursor structure for lacto/neolacto-series glycosphingolipids
(GSLs) in in vitro experiments. Here, we generated
B3gnt5-deficient (B3gnt5−/−) mice to investigate the in vivo biological
functions of lacto/neolacto-series GSLs. In biochemical analyses,
lacto/neolacto-series GSLs were confirmed to be absent and no
Lc3Cer synthase activity was detected in the tissues of these mice.
These results demonstrate that β3GnT5 is the sole enzyme synthesizing
Lc3Cer in vivo. Ganglioside GM1, known as a glycosphingolipid-
enriched microdomain (GEM) marker, was found to be upregulated
in B3gnt5−/− B cells by flow cytometry and fluorescence
microscopy. However, no difference in the amount of GM1 was observed
by TLC-immunoblotting analysis. The GEM-stained puncta on
the surface of B3gnt5−/− resting B cells were brighter and larger
than those of WT cells. These results suggest that structural alteration
of GEM occurs in B3gnt5−/− B cells.We next examinedwhether
BCR signaling-related proteins, such as BCR, CD19, and the signaling
molecule Lyn, had moved into or out of the GEM fraction. In
B3gnt5−/− B cells, thesemolecules were enriched in the GEMfraction
or adjacent fraction. Moreover, B3gnt5−/− B cells were more sensitive
to the induction of intracellular phosphorylation signals on BCR
stimulation and proliferated more vigorously than WT B cells. Together,
these results suggest that lacto/neolacto-series GSLs play
an important role in clustering of GEMs and tether-specific proteins,
such as BCR, CD19, and related signaling molecules to the GEMs..
18. Nishie T, Hikimochi Y, Zama K., Fukusumi Y, Ito M, Yokoyama H, Baruse C, Ito
M, Asano M.
, β-Galactosyltransferase-5 is a lactosylceramide synthase essential for
mouse extra-embryonic development.
, Glycobiology, 20, 10, 1311-1322, 2010.05.
19. 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.
20. Ishibashi Y, Nagamatsu Y, Meyer S, Imamura A, Ishida H, Kiso M, Okino N, Geyer R, Ito M., Transglycosylation-based Fluorescent Labeling of 6-Gala Series Glycolipids by EGALC, Glycobiology , 19, 7, 797-807, 2009.04.
21. Zama K, Hayashi Y, Ito Y, Hirabayashi Y, Inoue T, Ohno K, Okino N, Ito M., Simultaneous quantification of glucosylceramide and galactosylceramide by normal-phase HPLC using O-phtalaldehyde derivatives prepared with sphingolipid ceramide N-deacylase, Glycobiology , 19, 7, 767-775, 2009.05.
22. Kiyohara M, Sakaguchi K, Yamaguchi K, Araki T, and Ito M., Characterization and application of carbohydrate-binding modules of b-1,3-xylanase XUL4., J Biochem, 146, 5, 633-641, 2009.07.
23. Chisada S, Yoshimura Y, Sakaguchi K, Uchima H, Matsunaga N, Okino N, Uemura S, Go S, Ogura K, Tai T, Ikeda K, Taguchi R, Inokuchi J, and Ito M., Zebrafish and mouse a2,3-sialyltransferases responsible for synthesizing GM4 ganglioside., J Biol Chem., 284, 44, 30534-30546, doi:10.1074/jbc.M109.016188, 2009.10.
24. Sumida T., Ishii R, Yanagisawa T., Yokoyama S., and Ito M., Molecular cloning and crystal structural analysis of a novel b-N-acetylhexosaminidase from Paenibacillus sp. TS12 capable of degrading glycosphingoliids., J Mol Biol , 392, 87-99, 2009.04.
25. Xu X, Horibata Y, Inagaki M, Hama Y, Sakaguchi K, Okino N, and Ito M., A novel fucosyl glycosphingolipid of brine shrimp that is highly sensitive to endoglycoceramidase., Glycobiology , 19, 1446-1451, 2009.10.
26. Y. Hayashi, K. Zama, E. Abe, N. Okino, T. Inoue, K. Ohono, and M. Ito, A sensitive and reproducible fluorescent-based HPLC assay to measure the activity of acid as well as neutral b-glucocerebraosidases., Analytical Biochemistry, 383, 122-129, 2008.05.
27. H. M. Goda, K. Ushigusa, N. Okino, H. Narimatsu, and M. Ito, Molecular cloning, expression and characterization of a novel endo-a-N-acetylgalactosaminidase from Enterococcus faecalis., Biochemistry Biophysics Research Communication, 375, 541-546, 2008.05.
28. Y. Ishibashi, T. Nakasone, M. Kiyohara, K. Sakaguchi, A. Hijikata, N. Okino, M. Ito, A novel endogalactosylceramidase hydrolyzes oligogalactosylceramides to produce galactooligosaccharides and ceramides., Journal of Biological Chemistry, 282, 11386-11389, 2007.05.
29. Y. Hayashi, N. Okino, Y. Kakuta, T. Shikanai, H. Narimatsu, M. Ito, Klotho-related protein is a novel cytosolic neutral b-glycosylceramidase., Journal of Biological Chemistry, 282, 30889-30900, 2007.10.
30. Eriko Abe, Yasuhiro Hayashi, Yoichiro Hama, Masahiro Hayashi, Masanori Inagaki, and Makoto Ito, A novel phosphatidylcholine which contains pentadecanoic acid at sn-1 and docosahexaenoic acid at sn-2 in Shizochytrium sp. F26-b., Journal of Biochemistry, 140, 247-253, 2006.01.
31. Motohiro Tani, Yasuyuki Igarashi and Makoto Ito, Involvement of neutral ceramidase in ceramide metabolism at the plasma membrane and in extracellular milieu, Journal of Biological Chemistry, 10.1074/jbc.M506827200, 280, 44, 36592-36600, 280(44),36592-36600, 2005.11.
32. Shin-ichi Chisada, Yasuhiro Horibata, Yoichiro Hama, Masanori Inagaki, Naruto Furuya, Nozomu Okino and Makoto Ito, The glycosphingolipid receptor for Vibrio trachuri in the red sea bream intestine is a GM4 ganglioside which contains 2-hydroxy fatty acids., Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2005.05.110, 333, 2, 367-373, 333, 367-373, 2005.01.
33. Tetsuto Nakagawa, Akio Morotomi, Motohiro Tani, Noriyuki Sueyoshi, Hironobu Komori, and Makoto Ito, C18:3-GM1a induces appoptosis in neuro2a cells: enzymatic remodeling of fatty acyl chains of glycosphingolipids, Journal of Lipid Reserch, 10.1194/jlr.M400516-JLR200, 46, 6, 1103-1112, 46, 1103-1112, 2005.01.
34. Motohiro Tani, Nozomu Okino, Noriyuki Sueyoshi, and Makoto Ito, Conserved amino acid residues in the COOH-terminal tail are indispenzable for the correct folding and localization and enzyme activity of neutral ceramidase., Journal of Biological Chemistry, 10.1074/jbc.M404012200, 279, 28, 29351-29358, 2004.07.
35. Yukihiro Yoshimura, Motohiro Tani, Nozomu Okino, Hiroshi Iida, and Makoto Ito, Molecular cloning and functional analysis of zebrafish neutral ceramidase., Journal of Biological Chemistry, 10.1074/jbc.M405598200, 279, 42, 44012-44022, 279(42), 44012-44022, 2004.10.
36. Yasuhiro Horibata, Keishi Sakaguchi, Nozomu Okino, Hiroshi Iida, Masanori Inagaki, Yoichiro Hama and Makoto Ito, Unique catabolic pathway of glycosphingolipids in a Hydrozoan, Hydra magnipapillata, involving endoglycoceramidase, Journal of Biological Chemistry, 10.1074/jbc.M401460200, 279, 32, 33379-33389, 279(32), 33379-33389, 2004.08.
37. Motohiro Tani, Hiroshi Iida, Makoto Ito, O-Glycosylation of mucin-like domain retains the neutral ceamidase on the plasma membranes as a type II integral membrane protein, Journal of Biological Chemistry, 10.1074/jbc.M207932200, 278, 12, 10523-10530, 278 (12), 10523-10530, 2003.03.
38. Masako Furusato, Noriyuki Sueyoshi, Susumu Mitsutake, Makoto Ito et. al., Molecular cloning and characterization of sphingolipid ceramide N-deacylase from a marine bacterium, Shewanella alga G8, Journal of Biological Chemistry, 10.1074/jbc.m110688200, 277, 19, 17300-17307, 277(19), 17300-17307, 2002.05.
39. Susumu Mitsutake, Motohiro Tani, Nozomu Okino, Makoto Ito et al., Purification, characterization, molecular cloning , and subcellular distribution of neutral ceramidase of rat kidney., Journal of Biological Chemistry, 10.1074/jbc.M102233200, 276, 28, 26249-26259, 276(28), 26249-26259, 2001.07.
40. Motohiro Tani, Nozomu Okino, Susumu Mitsutake, Makoto Ito et al., Purification and characterization of a neutral ceamidase from mouse liver: A single protein catalyzes the reversile reaction in which ceramide is both hydrolyzed and synthesized, Journal of Biological Chemistry, 10.1074/jbc.275.5.3462, 275, 5, 3462-3468, 275(5), 3462-3468, 2000.02.
41. Motohiro Tani, Nozomu Okino, Kaoru Mori, Makoto Ito et al., Molecular cloning ot the full-length cDNA encoding mouse neutral ceamidase: A novel but highly conserved gene family of neutral ceramidase, Journal of Biological Chemistry, 10.1074/jbc.275.15.11229, 275, 15, 11229-11234, 275(15), 11229-11234, 2000.04.
42. Yasuhiro Horibata, Nozomu Okino, Sachiyo Ichinose, Akira Omori, Makoto Ito, Purification, characteriation, and cDNA cloning of a novel acidic endoglycoceramidase from the jellyfish, Cyanea nozakii, Journal of Biological Chemistry, 10.1074/jbc.M003575200, 275, 40, 31297-31304, 275(40), 31297-31304, 2000.10.
43. Hironobu Komori, Shinichi Ichikawa, Yoshio Hirabayashi, and Makoto Ito, Regulation of intracelular ceramide content in B16 melanoma cells: Biological implications of ceramide glycosylation, Journal of Biological Chemistry, 10.1074/jbc.274.13.8981, 274, 13, 8981-8987, 274(13), 8981-8987, 1999.03.
44. Nozomu Okino, Motohiro Tani, Shuhei Imayama, and Makoto Ito, Purification and characterization of a novel ceramidase from Pseudomonas aeruginosa, Journal of Biological Chemistry, 10.1074/jbc.273.23.14368, 273, 23, 14368-14373, 273(23), 14368-14373, 1999.06.
1. Makoto Ito, • Biological significance of sterylglucoside metabolism in Cryptococcus neoformans and Saccharomyces cerevisiae., International Conference of Lipid Bioscience (ICBL), 2015.09.
2. Takashi Watanabe, Makoto Ito, Integral roles of endoglycoceramidase-related protein 2 (EGCrP2) in pathogenic fungi Cryptococcus neoformans.
, International Conference of Lipid Bioscience (ICBL), 2014.06.
3. Ryo Sakiyama, Makoto Ito, LDRP1 regulates the triacylglycerol metabolism in thraustochytrids., International Conference of Lipid Bioscience (ICBL), 2014.06.
4. Ikumi Endo, Makoto Ito, Unique thraustochytrid steryl-β-glucosides and the gene responsible for their synthesis.
, International Conference of Lipid Bioscience (ICBL), 2014.06.
5. Makoto Ito, Shinya Uno, Eriko Abe, Takahiro Mizobuchi, Yuki Yamamoto, NOZOMU OKINO, Synthesis of DHA and DHA-containing glycerolipids and their accumulation in lipid droplets of thraustochytrids, 54th International Conference on the Bioscience of Lipids, 2013.09, Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) and their metabolites have attracted increasing attention in the development of medicines and nutritional supplements. Some of the primary producers of DHA in marine environments are thraustochytrids, which are protists capable of accumulating large amounts of DHA in lipid droplets (LDs). Recently, we showed a versatile transformation system (Sakaguchi et al. Appl Environ Microbiol, 2012), two distinct active pathways for DHA synthesis (Matsuda et al. J Lipid Res, 2012), and the increase of EPA production by delta 5 desaturase overexpression in thraustochytrids (Kobayashi et al. Appl Environ Microbiol, 2011). We previously reported that DHA made up 50 and 30% of fatty acids of phosphatidylcholine (PC) and triacylglycerol (TG), respectively (Abe et al. J Biochem, 2006); however, the mechanism of how DHA is incorporated into lipids at this ratio remains unknown. Here, we report a possible mechanism in which a novel phosphatidic acid (PA)-synthesizing enzyme, PLAT6, is involved. We cloned PLAT6 from Aurantiochytrium sp. as a novel lysophospholipid acyl transferase, and examined the specificity using the recombinant enzyme expressed in budding yeasts. PLAT6 was capable of catalyzing the specific transfer of DHA from DHA-CoA to lyso-PA, generating DHA-containing PA, which is a precursor for the synthesis of TG and PC in thraustochytrids. We also generated two mutant thraustochytrids, which exhibit abnormal phenotypes of LD formation, by targeted gene disruption: an mhdgat2-disruption mutant and ldrp1-disruption mutant lacking the conversion of diacylglycerol to TG and regulation of the LD size, respectively. The transformation system, novel genes, and mutants shown in this study will advance understanding of the global lipid metabolism of thraustochytrids, which will accelerate the industrial application of these promising single-cell organisms. .
6. 伊東 信, Lipids in Single Cell Organism: Unitiy in Diversity and Diversity in Unity, RIKEN Symposium, Evolution of Lipids, 2012.11.