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Production of a recombinant chitin deacetylase in the culture medium of Escherichia coli cells
Author(s) -
Tokuyasu Ken,
Kaneko Satoshi,
Hayashi Kiyoshi,
Mori Yutaka
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01113-8
Subject(s) - recombinant dna , escherichia coli , chitinase , chitin , biochemistry , signal peptide , biology , microbiology and biotechnology , enzyme , chitosan , gene
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.