| Project/Area Number |
08556015
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
ITO Makoto Kyushu University, Agriculture, Associate Professor, 農学部, 助教授 (40253512)
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| Co-Investigator(Kenkyū-buntansha) |
IZU Hiroyuki Takara Shuzo Co., Researcher, バイオ研究所, 研究員
HIGASHI Hideyoshi Mitsubishi Institute of Life Science, Senior researcher, 生命科学研究所, 主任研究員
OKINO Nozomu Kyushu University, Graduate School Division of Agriculture, Research Fellow of t, 大学院・農学研究科, 日本学術振興会特別研
佐野 睦 宝酒造バイオ研究所, 次席研究員
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥18,700,000 (Direct Cost: ¥18,700,000)
Fiscal Year 1997: ¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1996: ¥12,000,000 (Direct Cost: ¥12,000,000)
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| Keywords | Endoglycoceramidase / GM1 / Oligosaccharide / SCDase / Neuron / Apoptosis / Cell differentiation / Neurite / エンドブリコセラミダーゼ / 分化 / 糖脂質 / スフィンゴ脂質 / リゾ糖脂質 / EGCase / 逆反応 |
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| Research Abstract |
(1) The method for preparation of monosialoganglioside GM1 using sialidase-producing marine bacteria as a microbial biocatalyst was developed. A new sialidase-producing bacterium, identified as Pseudomonas sp. YF-2, was isolated from seawater. When YF-2 was cultured in a synthetic medium containing crude bovine brain gangliosides at 25ﾟC for 3 days, 80-90% of gangliosides were converted to GM1. The GM1 obtained was found to induceneurite outgrowth of neuroblastoma Neuro2a cells at a concentration of 33-100muM in the presence of fetal calf serum.
(2) The synthesis of [^<14>C]-labeled glycosphingolipids using the reverse hydrolysis reaction (condensation) of sphingolipid ceramide N-deacylase was developed. The optimum concentration of lyso-glycosphingolipids was 100-400muM depending on the species of lyso-form when[^<14>C]-stearic acid was used at the concentration of 100muM.Free[^<14>C]-fatty acids and lyso-glycosphingolipids were separated from the synthesized[^<14>C]-glycosphingolipids by using a Sep-Pak Plus Silica and a Sep-Pak CM or a QMA cartridge, respectively.
(3) The new method for preparation of a D-erythro SPC,which is natural occurring but difficult to be prepared by chemical methods, using marine bacteria as a biocatalyst was establised. We found for the first time that D-erythro SPC induced an apoptosis for human leukemia cell line HL 60 cells at a concentration of 50 muM.
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