Production and secretion of a recombinant Vibrio parahaemolyticus chitinase by Escherichia coli and its purification from the culture medium.
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.
Kadokura
Kazunari
K
Department of Biological Chemistry, College of Bioresource Sciences, Nihon University, Fujisawa, Kanagawa 252-8510, Japan.
Sakamoto
Yusuke
Y
Saito
Kaori
K
Ikegami
Takanori
T
Hirano
Takako
T
Hakamata
Wataru
W
Oku
Tadatake
T
Nishio
Toshiyuki
T
D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D002688:Chitinases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D016680:Genome, Bacterial; D016366:Open Reading Frames / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D014170:Transformation, Genetic; D014736:Vibrio parahaemolyticus / Q000201:enzymology* / Q000235:genetics
D001426:Bacterial Proteins; D003470:Culture Media; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D002688:Chitinases