Chitinase A from Stenotrophomonas maltophilia shows transglycosylation and antifungal activities

• Published in: Bioresource technology Vol. 133; pp. 213 - 220
• Main Authors: Suma, Katta, Podile, Appa Rao
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.04.2013; Elsevier
• Subjects: Antifungal activity; Antifungal Agents - pharmacology; antifungal properties; Biological and medical sciences; Chitin - metabolism; Chitinase; Chitinases - chemistry; Chitinases - isolation & purification; Chitinases - metabolism; Chitinases - ultrastructure; Chitooligosaccharides; Cloning, Molecular; Enzyme Stability - drug effects; Enzymes; Escherichia coli; Fundamental and applied biological sciences. Psychology; Fungi - drug effects; Fusarium oxysporum; Genes; Glycosylation - drug effects; Hydrogen-Ion Concentration - drug effects; Hydrolysis - drug effects; Kinetics; Microbial Sensitivity Tests; molecular cloning; Optimization; Polymers; Protein Structure, Tertiary; proteins; Sequence Analysis, Protein; Solubility; Stenotrophomonas maltophilia; Stenotrophomonas maltophilia - enzymology; Substrate Specificity - drug effects; Temperature; Time Factors; Transglycosylation; Stenotrophomonas maltophilia; Chitinase; Transglycosylation; Antifungal activity; Chitooligosaccharides; Pseudomonadales; Enzyme; Oligosaccharide; Biological activity; Glycosylases; Antifungal agent; Glycosidases; Bacteria; Hydrolases; Pseudomonadaceae
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2013.01.103

► Multi domain chitinase A and a single catalytic domain chitinase B of Stenotrophomonas maltophilia were characterized. ► Chitinase A was an endo acting enzyme active on both oligomeric and polymeric substrates. ► Chitinase B was exo/endo acting enzyme, that can hydrolyze only oligomeric substrates. ► Chitinase A, and not chitinase B, exhibited transglycosylation on tetra- and hexameric chitooligosaccharides. ► Chitinase A also showed antifungal activity against Fusarium oxysporum. Stenotrophomonas maltophilia chitinase (StmChiA and StmChiB) genes were cloned and expressed as soluble proteins of 70.5 and 41.6kDa in Escherichia coli. Ni–NTA affinity purified StmChiA and StmChiB were optimally active at pH 5.0 and 7.0, respectively and exhibited broad range pH activity. StmChiA and StmChiB had an optimum temperature of 40°C and are stable up to 50 and 40°C, respectively. Hydrolytic activity on chitooligosaccharides indicated that StmChiA was an endo-acting enzyme releasing chitobiose and StmChiB was both exo/endo-acting enzyme with the release of GlcNAc as the final product. StmChiA showed higher preference to β-chitin and exhibited transglycosylation on even chain length tetra- and hexameric substrates. StmChiA, and not StmChiB, was active on chitinous polymers and showed antifungal activity against Fusarium oxysporum.