Thermostable, salt tolerant, wide PH range novel chitobiase

Details Thermostable, salt tolerant, wide PH range novel chitobiase Thermostable, salt tolerant, wide PH range novel chitobiase

1993-12-21

1997-02-11

Wax, Robert A.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

435200, 435 691, 4351723, 4353201, 435 712, 4352523, 43525233, 536232, C12N 942, C12N 924, C12N 1556, C12N 1563

Patent

active

056020201

ABSTRACT:
A cloned chitobiase from a Vibrio parahemolyticus gene cloned into the plasmid pUC18 in E. coli strain DH5.alpha.. The plasmid construct, called pC120, had a 604 kb DNA insert. The recombinant gene expressed chitobiase activity similar to that found in native V. parahemolyticus. In addition to chitobiose, at least six additional substrates were observed to be hydrolyzed by the recombinant chitobiase, including .beta.-N-acetyl galactosamine glycosides, showing that the enzyme is an N-acetyl-hexosaminidase. The enzyme showed resistance to denaturation by 2M NaCl, was thermostable at 45.degree. C., and possessed an unusual range of activity from pH 5 to 9. The enzyme is useful in the degradation of crustacean shells. It catalyzes the production of N-acetyl-glucosamine, a compound which should be valuable as a chiral precursor or intermediate in the synthesis or manufacture of pharmaceutical compounds.

REFERENCES:
B. C. R. Zhu et al. FASEB J. 5(6) A1507 Abstract #6589 (Apr. 1991).
Zhu, B. C-R. "Chitobiase/B-N-Acetylhexosaminidase from Vibrio Parhemolyhzus; Salt Tolerant and Wide pH Range of Activity-Isolation, Characterization and Molecular Cloning." FASEB J. 5(6) A1507 Abstract #6589 (Apr. 1991).
Jannatipour et al., "Translocation of Vibrio harveyi N,N'-Diacetylchitobiase to the Outer Membrance of Escherichia coli," J. Bacteriol., vol. 169, pp. 3785-3791 (1987).
Soto-Gil et al., "N,N'-Diacetylchitobiase of Vibrio harveyi,"J. Biol. Chem. vol. 264, No. 25, pp. 14,778-14,783 (1989).
Wortman, et al., "Chitinase Determinants of Vibrio vulnificus: Gene Cloning and Applications of a Chitinase Probe," Appl. Environ. Microbiol., vol. 52, No. 1, pp. 142-145 (1986).
Kless et al., "Cloning of the Gene Coding for Chitobiase of Serratia marcescens,"Mol. Gen. Genet., vol. 217, pp. 471-473 (1989).
Facsimile letter from Roger Laine to Scott Davis (Feb. 13, 1991).

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