Degradation of Chitin and Chitosan by a Recombinant Chitinase Derived from a Virulent <i>Aeromonas hydrophila</i> Isolated from Diseased Channel Catfish — Oak Academic Publishing
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Degradation of Chitin and Chitosan by a Recombinant Chitinase Derived from a Virulent <i>Aeromonas hydrophila</i> Isolated from Diseased Channel Catfish
Aquatic Animal Health Research Unit, USDA-ARS, Auburn, USA
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Southern Regional Research Center, USDA-ARS, New Orleans, USA
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Aquatic Animal Health Research Unit, USDA-ARS, Auburn, USA
,
Southern Regional Research Center, USDA-ARS, New Orleans, USA
1 Aquatic Animal Health Research Unit, USDA-ARS, Auburn, USA
2 Southern Regional Research Center, USDA-ARS, New Orleans, USA
3 Aquatic Animal Health Research Unit, USDA-ARS, Auburn, USA
4 Southern Regional Research Center, USDA-ARS, New Orleans, USA
A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish ( Ictalurus punctatus ). Recombinant chitinase (rChi-Ah) was produced in Escherichia coli . Purified rChi-Ah had optimal activity at temperature of 42 ℃ and pH 6.5. The affinity ( Km ) for chitosan was 4.18 mg ·ml -1 with Vmax of 202.5 mg·min -1 ·mg -1 . With colloidal chitin as substrate, rChi-Ah generated N,N’-diacetyl-glucosamine predominantly. Conversion of chitosan (≥75% deacetylated) by rChi-Ah revealed five major products: 2 to 4 units of glucosamine, all of which had at least one acetyl group. It was determined that N-acetylated glucosamine was the recognition and cleavage site of rChi-Ah; the minimal and maximal cleavages were two and four glucosamine units, respectively. Functional analysis of rChi-Ah suggests that A. hydrophila chitinase is a bioactive chitinolytic enzyme, which may benefit the pathogen for survival and/or infection.
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