Cloning and characterization of two genes from Bacillus polymyxa expressing. beta. -glucosidase activity in Escherichia coli

Journal Article · Fri Dec 01 04:00:00 EST 1989 · Applied and Environmental Microbiology; (USA)

OSTI ID:5948478

Gonzalez-Candelas, L; Aristoy, M C; Polaina, J; Flors, A ^[1]

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1. Consejo Superior de Investigaciones Cientificas, Valencia (Spain)

DNA fragments from Bacillus polymyxa which encode {beta}-glucosidase activity were cloned in Escherichia coli by selection of yellow transformants able to hydrolyze the artificial chromogenic substrate p-nitrophenyl-{beta}-D-glucopyranoside. Restriction endonuclease maps and Southern analysis of the cloned fragments showed the existence of two different genes. Expression of either one of these genes allowed growth of E. coli in minimal medium with cellobiose as the only carbon source. One of the two enzymes was found in the periplasm of E. coli, hydrolyzed arylglucosides more actively than cellobiose, and rendered glucose as the only product upon cellobiose hydrolysis. The other enzyme was located in the cytoplasm, was more active toward cellobiose, and hydrolyzed this disaccharide, yielding glucose and another, unidentified compound, probably a phosphorylated sugar.

• OSTI ID: 5948478
• Journal Information: Applied and Environmental Microbiology; (USA), Journal Name: Applied and Environmental Microbiology; (USA) Vol. 55:12; ISSN 0099-2240; ISSN AEMID
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
BACILLUS
BACTERIA
CARBOHYDRATES
CELLOBIOSE
CLONING
DISACCHARIDES
DNA HYBRIDIZATION
DNA-CLONING
ENZYMES
ESCHERICHIA COLI
GENETIC ENGINEERING
GLUCOSIDASE
GLYCOSYL HYDROLASES
HYBRIDIZATION
HYDROLASES
METABOLISM
MICROORGANISMS
O-GLYCOSYL HYDROLASES
OLIGOSACCHARIDES
ORGANIC COMPOUNDS
SACCHARIDES