Cloning and Expression of Endo-β-1,3-glucanase gene from Flavobacterium dormitatov in Escherichia coli and Characterization of the Gene Product(Biological Chemistry)

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概要

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• The β-1,3-glucanase (1,3-β-D-glucan glucanohydrolase, EC 3.2.1.6) gene from Flavobacterium dormitator var. glucanolyticae was cloned into Escherichia coli C600 with a vector plasmid, pBR322. The E. coli cells carrying a recombinant plasmid, pKUβGl (8.2kb), showed a high β-1,3-glucanase activity and a lytic activity on viable yeast cells. These activities were found in the periplasmic space of E. coli clone cells. Southern hybridization analysis showed that the cloned gene was derived from F. dormitator chromosomal DNA. The gene products were purified from the periplasmic fraction of E, coli by ammonium sulfate fractionation and ion-exchange chromatography. The purified enzymes were demonstrated to be identical with a lytic endo-β-1,3-glucanase II and a nonlytic endo-β-1,3-glucanase I from F. dormitator from their enzymological and immunological properties. In the E. coli cells, endo-β-1,3-glucanase I was also formed by a proteolytic digestion of endo-β-1,3-glucanase II during the cultivation as in F. dormitator. Thus, the only endo-β-1,3-glucanase II was coded for in the cloned gene.

• 1990-10-23

著者

• Meguro H

  Tohoku Fukushi University

• Meguro H

  Department Of Applied Biological Chemistry Faculty Of Agriculture Tohoku University

• Misono Haruo

  Department Of Bioresources Science Faculty Of Agriculture Kochi University

• Misono Haruo

  Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kochi University

• NAGATA Shinji

  Laboratory of Bioorganic Chemistry, Department of Applied Biological Chemistry, Graduate School of A

• Maru Isafumi

  Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kochi University

• Nagasaki S

  Department Of Bioresources Science Kochi University

• Nagata Shinji

  Laboratory Of Applied Microbiology Department Of Bioresources Science Kochi University

• Nagasaki Susumu

  Laboratory of Applied Microbiology, Department of Bioresources Science, Kochi University

• ISHIHARA Fumiaki

  Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kochi University

• Moriyama Hideaki

  Japan Synchrotron Radiation Research Institute (JASRI)

• Ishihara Fumiaki

  Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kochi University

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