Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1995-05-31
1997-12-02
Prouty, Rebecca E.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435200, 4351723, 4352523, 43525233, 4353201, 536 231, 536 232, C12N 1556, C12N 1570, C12N 121, C12N 942
Patent
active
056935196
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 6.4 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:
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. 14,778-783 (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).
Zhu et al., "Chitobiase/B-N-Acetylhexosaminidase from Vibrio Parahemolyticus: Salt Tolerant and Wide pH Range of Activity--Isolation, Characterization and Molecular Cloning," FASEB J., 5(6) A1507 Abstract #6589 (Apr. 1991).
B.C.R. Zhu et al. "Chitobiase/B-N-Acetylhexosaminidase From Vibrio Parahemolyticus: Salt Tolerant and Wide pH Range of Activity--Isolation, Characterization and Molecular Cloning", FASEB J.5(6) A1507, Abst. #6589 1991.
Laine Roger A.
Ou Chin-Yih
Board of Supervisors of Louisiana State University and Agricultu
Prouty Rebecca E.
Runnels John H.
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