Molecular Cloning, Overexpression, and Purification of a Major Xylanase from Aspergillus oryzae

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概要

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• The gene encoding xylanase G2 (xynG2) was isolated from a genomic library of Aspergillus oryzae KBN616,used for making shoyu koji. The structural part of xynG2 was found to be 767 bp. The nucleotide sequence of cDNA amplified by RT-PCR showed that the open reading frame of xynG2 was interrupted by a single intron which was 71 bp in size and encoded 232 amino acids. Direct N-terminal amino acid sequencing showed that the precursor of XynG2 had a signal peptide of 44 amino acids. The predicted amino acid sequence of XynG2 has strong similarity to other family 11 xylanases from fungi. The xynG2 gene was successfully overexpressed in A. oryzae and the overpexpressed XynG2 was purified. The molecular weight of XynG2 estimated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 21,000. This was almost the same as the molecular weight of 20,047 calculated from the deduced amino acid sequence. The purified XynG2 showed an optimum activity at pH 6.0 and 58℃. It had a Km of 5.1 mg/ml and a Vmax of 123 μmol/min/mg when birch wood xylan was used as a substrate.

• 社団法人日本農芸化学会の論文
• 2000-12-23

著者

• Ohmiya Kunio

  Faculty Of Bioresources Mie University

• Araki T

  Faculty Of Bioresources Mie University

• Arai Takamitsu

  Faculty Of Bioresources And Center For Molecular Biology And Genetics Mie University

• KIMURA Tetsuya

  Faculty of Bioresources, Mie University

• Kajino T

  Special Research Laboratory Ii Toyota Central R&d Laboratories Inc.

• Morimoto K

  Faculty Of Agriculture Kagawa University

• Aburatani Takeshi

  Faculty Of Bioresources Mie University

• SUZUKI Hayato

  Faculty of Bioresources, Mie University

• FURUHASHI Hirofumi

  Faculty of Bioresources, Mie University

• MORIMOTO Kenji

  Faculty of Bioresources, Mie University

• SAKKA Kazuo

  Faculty of Bioresources, Mie University

• KARITA Shuichi

  Center for Molecular Biology and Genetics Mie University

• Morimoto Kenji

  Faculty Of Agriculture Kagawa University

• Morimoto Kenji

  Rare Sugar Research Center Faculty Of Agriculture Kagawa University

• Suzuki H

  Kirin Brewery Co. Ltd. Takasaki Jpn

• Sakka Kazuo

  Faculty Of Bioresources Mie University

• Arai T

  National Inst. Of Fruit Tree Sci. Nagasaki Jpn

• Kimura Tetsuya

  Graduate School Of Bioresources Mie University

• Furuhashi Hirofumi

  Faculty Of Bioresources Mie University

• Kimura Tetsuya

  Faculty Of Bioresources Mie University

• Suzuki Hayato

  Faculty of Bioresources Mie University

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