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Structural and functional analysis of insoluble-polysaccharide-hydrolyzing-enzyme by chimeric enzymes construction.

Research Project

Project/Area Number 07660093
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

OOI Toshihiko  Hokkaido Univ., Fac.of Eng., Assoc.Pro., 工学部, 助教授 (40223713)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsAspergillus aculeatus / Bacillus pumilus / cellulase / xylanase / chieric enzyme / glycosidase / Aspergillus aculeatus
Research Abstract

To understand the structure and function relationsip of the insoluble polysaccharide-hydrolyzing-enzymes, such as cellulase and xylanase, we planned the research that construct chimeric protein (enzyme) between cellulase of Aspergillus aculeatus and xylanase of Bacillus pumilus that based on both stereostructure of the enzymes. To produce the chimeric enzyme proteins, the genes of the both enzymes were used. The chemeric genes which named XCX and CXC,were constructed by using restriction site and/or PCR method. The constructed genes were identified by the nucleotide sequencing. Both genes were expressed in Escherichia coli. The chimeric proteins that expressed in E.coli were purified to homogeneity by conventional chromatographic tequnique. Both purified chimeric proteins were reacted to antiserum against cellulase and xylanase from original strains. The activity of the chimeric proteins were analyzed to sensitive substrates, which were 4-methylumbeiferyl-beta-D-cellobiose, cellotriose, and xylose, respectively. The activities of the chimeric proteins were less than ditectable level.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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