Heterologous Expression and Characterization of Endoglucanase I (EGI) from Trichoderma viride HK-75
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概要
- 論文の詳細を見る
Endoglucanase I (EGI) secreted from Trichoderma viride HK-75 has a unique transglycosylation activity. The genomic and cDNA clones encoding EGI (egl1) of T. viride HK-75 were isolated and characterized. The coding region of egl1,composed of 1392 bp, was found to encode a polypeptide of 464 amino acids that has extensive similarity (93.8%) with EGI of T. reesei. Expression of the egl1 gene in E. coli as a fusion protein (with N-terminal thioredoxin and C-terminal histidine tag) led to a large production of a nonglycosylated protein of 62.5 kDa. However, it formed an insoluble inclusion body. Upon denaturation with 8M urea followed by dialysis and successive purification, the enzymatically active recombinant EGI (rEGI) was obtained at a level as high as 18.3mg/l of 1,000ml of culture. The rEGI had 67.8% activity for carboxymethyl cellulose (CMC), compared to native EGI (nEGI). The optimum pH and optimum temperature of rEGI were lower than those of nEGI by 0.5 and 5℃, respectively. The rEGI also had narrower CMCase ranges than nEGI in pH and temperature stabilities. However, the catalytic and transglycosylation abilities against cellotriose of rEGI were comparable to those of nEGI. These results suggest that the glycosylation is important for the stabilities of EGI but not critical for the essential enzymatic capacity.
- 社団法人日本農芸化学会の論文
- 1999-10-23
著者
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Goto M
Univ. Tokyo Tokyo Jpn
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FURUKAWA Kensuke
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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Goto Masatoshi
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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Ekino K
Kyushu Univ. Fukuoka Jpn
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Ekino Keisuke
Department Of Bioscience And Biotechnology Faculty Of Agriculture Kyushu University
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Goto Masatoshi
Department Of Agricultural Chemistry Kyushu University
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Furukawa Kensuke
Division Of Bioresource And Bioenvironmental Sciences Kyushu University
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Furukawa Kensuke
Graduate School Of Bioresource And Bioenvironmental Sciences Kyushu University
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Goto Masatoshi
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kyushu University
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Furukawa K
Graduate School Of Bioresource And Bioenvironmental Sciences Kyushu University
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KWON Il
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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EKINO Keisuke
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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Kwon Il
Department Of Bioscience And Biotechnology Faculty Of Agriculture Kyushu University
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