Overexpression and Purification of Aspergillus aculeatus β-Mannosidase and Analysis of the Integrated Gene in Aspergillus oryzae

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概要

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• An expression plasmid for the manB gene encoding Aspergillus aculeatus β-D-mannosidase (MANB) was constructed by using an expression vector carrying an improved promoter. After transformation of A.oryzae by the plasmid, several transformants formed colonies emitting fluorescence on a plate containing 4-methylumbelliferyl β-D-mannopyranoside (MU-Man) under UV-irradiation. The transformant that displayed the strongest fluorescence, named A.oryzae BMN1, produced about 270 mg MANB/l in liquid culture. Recombinant MANB overproduced in BMN1 was purified by two steps of column chromatography to a single protein band on SDS-polyacrylamide gel electrophoresis and had a molecular weight of 130, 000. Analyses by Southern blotting and genomic PCR demonstrated that a single copy of the plasmid was integrated into the chromosome by recombination at the niaD locus.

• 社団法人日本生物工学会の論文
• 2001-08-25

著者

• Takada G

  Rare Sugar Research Center Kagawa University

• KAWAGUCHI TAKASHI

  Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University

• SUMITANI JUN-ICHI

  Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University

• ARAI MOTOO

  Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University

• Kawaguchi Takashi

  Department Of Applied Life Sciences Graduate School Of Life And Environmental Sciences Osaka Prefect

• Kawaguchi Takashi

  Department Of Applied Biological Chemistry College Of Agriculture Osaka Prefecture University

• Kawaguchi Takashi

  College Of Agriculture University Of Osaka Prefecture

• Kawaguchi Takashi

  Department Of Drug Safety Research Eisai Co. Ltd.

• Arai M

  Department Of Environmental Biology College Of Bioscience And Biotechnology Chubu University

• Arai Motoo

  Graduate School Of Agriculture And Biological Sciences Osaka Prefecture University

• KANAMASA Shin

  Graduate School of Life and Environmental Sciences, Osaka Prefecture University

• Kawaguchi T

  Department Of Applied Life Sciences Graduate School Of Life And Environmental Sciences Osaka Prefect

• Tajima Gou

  Department Of Pediatrics Hiroshima University Graduate School Of Biomedical Sciences

• Kanamasa Shin

  Graduate School Of Life And Environmental Sciences Osaka Prefecture University

• TAKADA GORO

  Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University

• Takata Goro

  Rare Sugar Research Center Kagawa University

• Sumitani J

  Graduate School Of Life And Environmental Sciences Osaka Prefecture University

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