Cloning and Sequencing of a Novel Endo-1,4-β-Glucanase Gene from Ruminococcus albus

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概要

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• The gene encoding an endo-1,4-β-glucanase from Ruminococcus albus F-40 was cloned in Escherichia coli JM109 using pBR322. The nucleotide sequence of the 1,798 bp PstI-PvuII fragement which includes a cellulase gene was determined. There as a single open reading frame (ORF) consisting of 936 bp encoding a peptide of 312 amino acid residues with a molecular weight of 35,760. The N-terminal amino acid sequence determined for the enzyme expressed in E. coli was identitical to that deduced from the beginning of the ORF. A putative ribosome-binding site and a promotor were located upstream of the ORF. Activity was expressed from this fragment when it was subcloned in both orientations in pUC118 and pUC119,indicating that its own promotor functioned in E. coli. The amino acid sequence of the endoglucanase deduced from the nucleotide sequence showed 44% homology with CelA from Butyrivibrio fibrisolvens A46,suggesting that this enzyme was a member of family A2. The enzyme purified from E. coli exhibited the highest activity against carboxymethyl cellulose (CMC) at 40℃ and pH around 7.0.

• 社団法人日本生物工学会の論文
• 1993-12-25

著者

• Kajino T

  Toyota Central Research & Development Labs. Inc.

• Kajino T

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

• Kimura Tetsuya

  National Center For Seeds And Seedlings

• Morimoto K

  Faculty Of Agriculture Kagawa University

• SAKKA Kazuo

  Applied Microbiology Laboratory, Graduate School of Bioresources, Mie University

• KARITA Shuichi

  Center for Molecular Biology and Genetics Mie University

• Morimoto Kenji

  Rare Sugar Research Center Faculty Of Agriculture Kagawa University

• MORIOKA KENSEI

  Applied Microbiology Laboratory, Faculty of Bioresources

• KAJINO TSUTOMU

  Mie University, Toyota Central Research and Development Laboratory Inc.,

• SHIMADA KYO

  Applied Microbiology Laboratory, Faculty of Bioresourses

• OHMIYA KUNIO

  Applied Microbiology Laboratory, Faculty of Bioresources

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