Production and secretion of a recombinant Vibrio parahaemolyticus chitinase by Escherichia coli and its purification from the culture medium

• Published in: Bioscience, biotechnology, and biochemistry Vol. 71; no. 11; pp. 2848 - 2851
• Main Authors: Kadokura, K.(Nihon Univ., Fujisawa, Kanagawa (Japan). Coll. of Bioresource Sciences), Sakamoto, Y, Saito, K, Ikegami, T, Hirano, T, Hakamata, W, Oku, T, Nishio, T
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.11.2007; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: ADN RECOMBINADO; ADN RECOMBINE; Bacterial Proteins - biosynthesis; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Biological and medical sciences; CHITINASE; Chitinases - biosynthesis; Chitinases - genetics; Chitinases - isolation & purification; Cloning, Molecular; Culture Media - chemistry; ESCHERICHIA COLI; Escherichia coli - genetics; FERMENTACION; FERMENTATION; Fundamental and applied biological sciences. Psychology; GENETICALLY MODIFIED MICROORGANISMS; Genome, Bacterial; IMMUNOTHERAPIE; IMMUNOTHERAPY; INMUNOTERAPIA; MICRO-ORGANISME MODIFIE GENETIQUE; MICROORGANISMOS MODIFICADOS GENETIC; Open Reading Frames - genetics; Protein Sorting Signals - genetics; QUITINASA; RECOMBINANT DNA; recombinant enzyme; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Recombinant Proteins - secretion; signal peptide; Transformation, Genetic; VIBRIO PARAHAEMOLYTICUS; Vibrio parahaemolyticus - enzymology; Vibrio parahaemolyticus - genetics; Culture medium; Leader peptide; Purification; Enzyme; Secretion; Escherichia coli; Vibrio parahaemolyticus; signal peptide; chitinase; Production; Bacteria; recombinant enzyme; Enterobacteriaceae
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.70389

An open reading frame encoding the chitinase gene and its signal sequence was cloned from the Vibrio parahaemolyticus KN1699 genome. An expression plasmid containing the gene was introduced into Escherichia coli cells, and recombinant chitinase (Pa-rChi) was produced and secreted into the culture medium with the aid of the signal peptide. Pa-rChi was purified and its substrate specificity was determined.