Development of thermostable protein-refolding catalysis and characterization of the kinetic properties
| Project/Area Number |
21580112
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied biochemistry
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| Research Institution | Okayama University |
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Principal Investigator |
TAMURA Takashi 岡山大学, 大学院・自然科学研究科, 准教授 (40253009)
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| Co-Investigator(Renkei-kenkyūsha) |
KURAMITSU Seiki 大阪大学, 大学院・理学部研究科, 教授 (60153368)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2010: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 酵素化学 / ジスルフィド架橋 / 蛋白質フォールディング / 酸化還元電位 / 平衡論支配 / 速度論支配 / シグナルペプチド配列 / ペリプラズム空間 / ジスルフィドイソメラーゼ / ジスルフイド架橋 / ジスルフィト架橋 / シグナルペブチド配列 / ペリープラズム空間 |
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| Research Abstract |
The present study assessed the contribution of sctive site sequence of Escherichia coli DsbA by screening a library of mutant dsbA genes with the genetically randomized active-site sequence CXXC. Alleles of 24 dsbA mutants gave significant variation in the cellular functions, and a mutant dsbA gene that encodes Cys-Asp-Ile-Cys sequence enhanced pahge sensitivity by 40-fold more than the wild-type dsbA. The redox potential of DsbA[CDIC] was determined to be. 171. 8 mV, which suggested less oxidizing catalyst than the wild type DsbA(-125. 9 mV) and the redox potential was close to that of yeast protein disulfide isomerase(175 mV). It was suggested that DsbA[CDIC] improved the net yield of the correctly folded periplasmic proteins through the disulfide isomerase catalysis. Thermostable homologs were also characterized for application of folding catalysis at high temperature.
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Report
(4 results)
Research Products
(41 results)
