Production of a recombinant chitin deacetylase in the culture medium of Escherichia coli cells

FEBS Letters

Mori, Yutaka¹  Hayashi, Kiyoshi¹  Tokuyasu, Ken¹  Kaneko, Satoshi¹ 

[1] National Food Research Institute, Kannondai 2-1-2, Tsukuba 305-8642, Japan

关键词: Chitin deacetylase;    Recombinant enzyme;    Colletotrichum lindemuthianum;    Escherichia coli;    CDA;    chitin deacetylase;   

DOI  :  10.1016/S0014-5793(99)01113-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

With the aid of a signal sequence of a chitinase from Streptomyces lividans, a recombinant chitin deacetylase, whose gene originated from a Deuteromycete, Colletotrichum lindemuthianum, was produced in the culture medium of Escherichia coli cells, existing as a highly active form without the signal peptide. During the production of the recombinant chitin deacetylase, both a slight increase in the value of OD_{600 nm} in the culture medium and a drastic decrease in viable cell number were observed. When penta-N-acetyl-chitopentaose was used as the substrate, the recombinant chitin deacetylase had comparable kinetic parameters to those of the original enzyme from the fungus. The addition of a C-terminal six histidine sequence to the recombinant enzyme caused a slight decrease in the k _cat value, and the further addition of a 12 amino acid sequence at its N-terminus caused a further decrease in the value. This production system allowed us to easily produce in the culture media the recombinant chitin deacetylases possessing as good properties as the original enzyme, without any disruption steps of the E. coli cells.

【 授权许可】

Unknown