A new [kappa]-carrageenase CgkS from marine bacterium Shewanella sp. Kz7

A new [kappa]-carrageenase gene cgkS was cloned from marine bacterium Shewanella sp. Kz7 by using degenerate and site-finding PCR. The gene was comprised of an open reading frame of 1224 bp, encoding 407 amino acid residues, with a signal peptide of 24 residues. Based on the deduced amino acid seque...

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Published inJournal of Ocean University of China. JOUC Vol. 14; no. 4; p. 759
Main Authors Wang, Linna, Li, Shangyong, Zhang, Shilong, Li, Jiejing, Yu, Wengong, Gong, Qianhong
Format Journal Article
LanguageEnglish
Published Dordrecht Springer Nature B.V 01.08.2015
Subjects
Amino acids
Bacteria
Carbohydrates
Cloning
E coli
Enzymes
Sodium chloride
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Summary:A new [kappa]-carrageenase gene cgkS was cloned from marine bacterium Shewanella sp. Kz7 by using degenerate and site-finding PCR. The gene was comprised of an open reading frame of 1224 bp, encoding 407 amino acid residues, with a signal peptide of 24 residues. Based on the deduced amino acid sequence, the [kappa]-carrageenase CgkS was classified into the Glycoside Hydrolase family 16. The cgkS gene was expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity with a specific activity of 716.8 U mg^sup -1^ and a yield of 69%. Recombinant CgkS was most active at 45°C and pH 8.0. It was stable at pH 6.0-9.0 and below 30°C. The enzyme did not require NaCl for activity, although its activity was enhanced by NaCl. CgkS degraded [kappa]-carrageenan in an endo-fashion releasing tetrasaccharides and disaccharides as main hydrolysis products.
ISSN:1672-5182
1672-5174
DOI:10.1007/s11802-015-2713-y