Cloning, Sequencing, and Expression of a β-Amylase Gene from Bacillus cereus var. mycoides and Characterization of Its Products

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概要

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• The cloned gene was composed of 1638 bp for coding plus promoter like and SD-like sequences ahead of it. The deduced amino acid sequence had high similarity with known β-amylases. The N-terminal sequence of the cloned β-amylase seemed to be a signal peptide. The gene was introduced into Bacillus subtilis 1A289 using pHY300PLK as a vector and the expressed protein was recovered from the culture media. The enzyme fraction produced was divided into two components upon the DEAE column chromatography. The amino acid sequence of one fraction (FrI) was the same as the mature enzyme, and the other (FrII) lacked the N-terminal amino acid residue (Ala) of the mature enzyme. The kinetic parameters of the hydrolysis catalyzed by the enzyme component fir were measured, and the subsite affinities of the enzyme were evaluated. In conclusion, it was shown that the recombinant enzyme was the same as the mature enzyme functionally and proteochemically.

• 社団法人日本農芸化学会の論文
• 1996-08-23

著者

• Kozaki S

  Laboratory Of Veterinary Public Health College Of Agriculture Osaka Prefecture University

• YAMAGUCHI TAICHI

  Laboratory of Human Performance and Fitness, Graduate School of Education, Hokkaido University

• Hibino Takeshi

  Laboratory Of Biophysical Chemistry College Of Agriculture Osaka Prefecture University

• Takasaki Yoshiyuki

  Department of Materials Science, Faculty of Engineering, Miyazaki University

• Yamaguchi Takayuki

  Department Of Bioresources Chemistry Faculty Of Horticulture Chiba University Engineering Department

• Yamaguchi Taichi

  Laboratory Of Biophysical Chemistry College Of Agriculture Osaka Prefecture University

• Takasaki Yoshiyuki

  Department Of Applied Chemistry Faculty Of Engineering Miyazaki University

• Takasaki Yoshiyuki

  Laboratory Of Biophysical Chemistry College Of Agriculture Osaka Prefecture University:dainippon Phe

• Yamaguchi T

  Department Biochemistry National Institute Of Agrobiological Science

• Matsumoto Yasuo

  Laboratory Of Biophysical Chemistry College Of Agriculture Osaka Prefecture University

• NITTA Yasunori

  Laboratory of Biophysical Chemistry, College of Agriculture, Osaka Prefecture University

• Nitta Y

  College Of Agriculture Osaka Prefecture University

• SHIRAKAWA Mayumi

  Laboratory of Biophysical Chemistry, College of Agriculture, Osaka Prefecture University

• KIBE Mayumi

  Laboratory of Biophysical Chemistry, College of Agriculture, Osaka Prefecture University

• KOZAKI Shunji

  Laboratory of Veterinary Public Health, College of Agriculture, Osaka Prefecture University

• SHIRAKAWA Mayumi

  Department of Materials Science, Faculty of Engineering, Miyazaki University

• Kibe Mayumi

  Laboratory Of Biophysical Chemistry College Of Agriculture Osaka Prefecture University

• Nitta Yasunori

  Laboratory Of Biophysical Chemistry College Of Agriculture Osaka Prefecture University

• Shirakawa Mayumi

  Department Of Materials Science Faculty Of Engineering Miyazaki University

• Kozaki Shunji

  Laboratory Of Veterinary Epidemiology Department Of Veterinary Science Graduate School Of Life And Environmental Sciences Osaka Prefecture University

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