Novel Bi-modular GH19 chitinase with broad pH stability from a fibrolytic intestinal symbiont of eisenia fetida, cellulosimicrobium funkei HY-13

Abstract

Endo-type chitinase is the principal enzyme involved in the breakdown of N-acetyl-D-glucosamine-based oligomeric and polymeric materials through hydrolysis. The gene (966-bp) en-coding a novel endo-type chitinase (ChiJ), which is comprised of an N-terminal chitin-binding domain type 3 and a C-terminal catalytic glycoside hydrolase family 19 domain, was identified from a fibrolytic intestinal symbiont of the earthworm Eisenia fetida, Cellulosimicrobium funkei HY-13. The highest endochitinase activity of the recombinant enzyme (rChiJ: 30.0 kDa) toward colloidal shrimp shell chitin was found at pH 5.5 and 55 °C and was considerably stable in a wide pH range (3.5– 11.0). The enzyme exhibited the highest biocatalytic activity (338.8 U/mg) toward ethylene glycol chitin, preferentially degrading chitin polymers in the following order: ethylene glycol chitin > col-loidal shrimp shell chitin > colloidal crab shell chitin. The enzymatic hydrolysis of N-acetyl-β-D-chitooligosaccharides with a degree of polymerization from two to six and colloidal shrimp shell chitin yielded primarily N,N′-diacetyl-β-D-chitobiose together with a small amount of N-acetyl-D-glucosamine. The high chitin-degrading ability of inverting rChiJ with broad pH stability suggests that it can be exploited as a suitable biocatalyst for the preparation of N,N′-diacetyl-β-D-chitobiose, which has been shown to alleviate metabolic dysfunction associated with type 2 diabetes.