| Project/Area Number |
14205112
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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 | 一般 |
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| Research Field |
化学工学一般
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| Research Institution | Kobe University |
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Principal Investigator |
FUKUDA Hideki Kobe University, Graduate School of Science and Technology, Professor, 大学院・自然科学研究科, 教授 (30263396)
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| Co-Investigator(Kenkyū-buntansha) |
KATOH Shigeo Kobe University, Graduate School of Science and Technology, Professor, 大学院・自然科学研究科, 教授 (20026272)
KONDO Akihiko Kobe University, Engineering, Professor, 工学部, 教授 (40205547)
ISHIKAWA Haruo Osaka Prefecture University, Graduate School of Engineering, Emeritus Professor, 大学院・工学研究科, 教授 (00081349)
OOSHIMA Hiroshi Osaka City University, Graduate School of Engineering, Professor, 大学院・工学研究科, 教授 (20112526)
OGINO Hiroyasu Osaka Prefecture University, Graduate School of Engineering, Lecturer, 大学院・工学研究科, 講師 (80233443)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥53,820,000 (Direct Cost: ¥41,400,000、Indirect Cost: ¥12,420,000)
Fiscal Year 2004: ¥11,440,000 (Direct Cost: ¥8,800,000、Indirect Cost: ¥2,640,000)
Fiscal Year 2003: ¥22,230,000 (Direct Cost: ¥17,100,000、Indirect Cost: ¥5,130,000)
Fiscal Year 2002: ¥20,150,000 (Direct Cost: ¥15,500,000、Indirect Cost: ¥4,650,000)
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| Keywords | cell-surface engineering / Flo1p anchor protein / ethanol production / biodiesel fuel / enantioselective reaction / solvent-tolerant enzyme / cell wall structure / immunoliposome / 表層提示用タンパク質Flo1 / グルコアミラーゼ / α-アミラーゼ / ヘミセルロース / キシロース / β-1,6-グルカン / リポソーム / ファージライブラリー / 凝集性遺伝子Flolp / 表層発現酵母 / ZZドメイン / GPIアンカー / 磁性ナノ粒子 / 凝集性遺伝子Flo1p / リパーゼ / アミラーゼ / エタノール醗酵 / キシラナーゼ / whole cell biocatalyst |
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| Research Abstract |
In this study, to construct yeast strains with high cell surface enzyme activity, a novel cell-surface display system, based on the FLO1 gene encoding a lectin-like cell-wall protein (Flo1p) in Saccharomyces cerevisiae, was investigated. Several topics obtained are as follows :
1.Direct and efficient productions of ethanol by fermentation from raw corn starch and cellulosic material were achieved by using the yeast S.cerevisiae condisplaying two amylolytic enzymes such as glucoamylase and α-amylase and three cellulolytic enzymes such as endoglucanase, cellobiohydrolase and β-glucosodase, respectively.
2.Using this system using Flo1p protein, recombinant lipase with a pro sequence from Rhizopus oryzae (ProROL) was displayed on the cell surface, and sucuessfully catalyzed the methanolysis reaction for biodiesel fuel production. Besides, ProROL displayed on the yeast cell surface could efficiently catalyze enantioselective transesterification in non-aqueous organic solvent.
3.A lipase gene (lip 3) was cloned from the Pseudomonas aeroginosa strain LST-03, which tolerates organic solvents and expressed in Eschrichi coli. Furtheremore, the relationship between LST-03 and Lip3 lipases was investigated. The kinetics and mechanism of the reaction catalyzed by the P.aeruginosa PST-01 protease, which was obtained from an organic solvent-tolerant microorganism P.aeruginosa, was clarified.
4.To explore the relationship between the GPI anchor structure and β-1,6-glucan synthesis, deletion mutants in genes involved in GPI synthesis for osmotic remedial growth were screened. The mcd4 deletion causes a decrease in GPI cell wall proteins levels, and the mutation caused a decrease in mannan levels and increase in alkali-insoluble β-1,6-glucan and chitin levels in the cell wall.
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