Molecular cloning, expression and characterization of a chitosanase from Microbacterium sp.
A gene encoding a chitosanase (mschito) was cloned from Microbacterium sp. OU01. The ORF consists of 801 bp which encoded a polypeptide of 266 amino acid residues. The deduced amino acid sequence shows 98% identity to that of the chitosanase reported in Pseudomonas sp. A-01. In addition, the fusion protein containing MSCHITO was expressed in E. coli and purified using Ni-NTA affinity chromatography. The purified rMSCHITO protein degraded the chitosan (the degree of deacetylation of 99%) and produced a mixture of chitooligosaccharides. The MSCHITO is thus an endo-chitosanase.
Zhang
Jiquan
J
Key Laboratory of Marine Biotechnology of Jiangsu Province, Huaihai Institute of Technology, Lianyungang, 222005, P.R. China.
Sun
Yuying
Y
D000595:Amino Acid Sequence; D001709:Biotechnology / Q000379:methods*; D048271:Chitosan / Q000737:chemistry; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D016678:Genome; D016680:Genome, Bacterial; D006026:Glycoside Hydrolases / Q000737:chemistry*; D006094:Gram-Positive Bacteria / Q000201:enzymology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D048271:Chitosan; D006026:Glycoside Hydrolases; C019868:chitosanase