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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제25권 제7호
- 발행연도
- 2015.1
- 수록면
- 988 - 998 (11page)
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초록· 키워드
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The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.
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참고문헌 (23)
참고문헌 신청
Akao T / 2006 / Cloning and expression of 1, 2-α-mannosidase gene (fmanIB) from filamentous fungus Aspergillus oryzae: in vivo visualization of the FmanIBp-GFP fusion protei
Balkan B / 2005 / Production and properties of α-amylase from Penicillium chrysogenum and its application in starch hydrolysis / Prep. Biochem. Biotechnol 35 : 169 ~ 178
Barbesgaard P / 1992 / On the safety of Aspergillus oryzae: a review / Appl. Microbiol. Biotechnol 36 : 569 ~ 572
Bradford MM / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72 :
Chalfie M. / 1995 / Green fluorescent protein / Photochem. Photobiol 62 : 651 ~ 656
Balkan B / 2005 / Production and properties of α-amylase from Penicillium chrysogenum and its application in starch hydrolysis / Prep. Biochem. Biotechnol 35 : 169 ~ 178
Barbesgaard P / 1992 / On the safety of Aspergillus oryzae: a review / Appl. Microbiol. Biotechnol 36 : 569 ~ 572
Bradford MM / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72 :
Chalfie M. / 1995 / Green fluorescent protein / Photochem. Photobiol 62 : 651 ~ 656
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