Molecular Cloning of Acid-Stable Glucose Isomerase Gene from Streptomyces olivaceoviridis E-86 by a Simple Two-Step PCR Method, and Its Expression in Escherichia coli

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概要

• 論文の詳細を見る

• Glucose isomerase (GI) from Streptomyces olivaceoviridis E-86 is a unique enzyme, very acid-stable with a large potential for corn sweetener industries. The gene encoding this unique enzyme was cloned by a simple two-step PCR method, and expressed in Escherichia coli. A single open reading frame consisting of 1164 base pairs (70.7 mol% of G+C content) that encoded a polypeptide composed of 388 amino acid residues (M_r 42,993) was found. The E.coli transformant carrying the gene overproduced the recombinant GI (rGI) and the enzyme was successfully expressed as a tetramer under the transcriptional control of the tac-promoter. The purified recombinant enzyme was indistinguishable from that of the authentic enzyme e.g. molecular weight, immunological properties, N-terminal amino acid sequences, subunit structures, and temperature and pH profiles. The relationships between structure and properties of the enzymes are also discussed.

• 社団法人日本農芸化学会の論文
• 2001-05-23

著者

• Takahashi S

  Kyoto Univ. Kyoto Jpn

• Saito K

  Department Of Kampo Medicinal Science Hokkaido College Of Pharmacy

• Kawamura Yukio

  Department Of Fermentation Science Faculty Of Applied Bio-science Tokyo University Of Agriculture

• Kawamura Y

  The Graduate School Of Science And Technology Kobe University

• Kawamura Y

  Department Of Bioengineering Akita Research Institute Offood And Brewing (arif)

• Takahashi K

  School Of Life Sciences Tokyo Univ. Of Pharmacy And Life Sciences

• Takahashi Kayoko

  Central Research Lnstitute Meiji Milk Products Co. Ltd.

• Takahashi S

  Department Of Biosciences Teikyo University

• Takahashi S

  Akita Res. Inst. Of Food And Brewing

• Takahashi Saori

  Akita Res. Inst. Of Food And Brewing

• Takahashi S

  Akita Res. Inst. Food And Brewing Akita

• Takahashi Shunji

  Institute Of Molecular And Cellular Biosciences University Of Tokyo

• Takahashi Shunji

  Department Of Bioresources Chemistry Faculty Of Horticulture Chiba University

• 斉藤 謙一

  Department Of Physiology And Anatomy Nihon University College Of Pharmacy

• TAKAHASHI Saori

  Department of Bioengineering, Akita Research Institute of Food and Brewing (ARIF)

• SAITO Kyuichi

  Department of Bioengineering, Akita Research Institute of Food and Brewing (ARIF)

• KANEKO Takahiro

  Department of Bioengineering, Akita Research Institute of Food and Brewing (ARIF)

• Kawamura Yukio

  Department Of Bioengineering Akita Research Institute Offood And Brewing (arif)

• Saguchi Ken-ichi

  Laboratory Of Biopharmaceutics School Of Pharmaceutical Sciences Showa University

• Shinozaki Kazuo

  Riken Kanagawa Jpn

• Takahashi Kyoko

  Department Of Food Science And Nutrition School Of Human Environmental Sciences Mukogawa Women'

• Sasaki Kanako

  Riken Kanagawa Jpn

• Kageyama T

  Kyoto Univ. Aichi Jpn

• Suzuki Takahito

  Biological Laboratory Faculty Of Science Nara Women's University

• Takahashi Saori

  Department Of Applied Biology Faculty Of Textile Science Kyoto Institute Of Technology

• Takahashi Kenji

  Laboratory Of Molecular Biochemistry School Of Life Science Tokyo University Of Pharmacy And Life Sc

• Kawamura Yukio

  Department Of Applied Biological Chemistry Graduate School Of Agriculture Kinki University

• Saito Kyuichi

  Department of Bioengineering Akita Research Institute of Food and Brewing (ARIF)

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