Cloning and expression in Escherichia coli of a gene coding for a chondroitin lyase from Bacteroides thetaiotaomicron

Journal Article · Fri Nov 01 04:00:00 EST 1985 · J. Bacteriol.; (United States)

OSTI ID:5448903

Guthrie, E P; Shoemaker, N B; Salyers, A A

The authors cloned the gene for one of the two chondroitin lyases of Bacteroides thetaiotaomicron into the cosmid vector pHC79 and subcloned it into pBR328. No proteins the size of B. thetaiotamicron chondroitin lyase I or II (104 to 108 kilodaltons) were detectable in maxicell or in vitro transcription-translation preparations. However, partial purification of the chondroitin lyase activity from the Escherichia coli subclone showed that its properties were similar to those of the B. thetaiotaomicron chondroitin lyases. Antibodies to the chondroitin lyase that was produced in E. coli cross-reacted with the B. thetaiotaomicron chondroitin lyase II but not with chondroitin lyase I. The molecular weight of the enzyme produced in E. coli was slightly lower than those of the two chondroitin lyases from B. thetaiotaomicron. The chondroitin lyase gene was located on the subcloned 7.8-kilobase EcoRI fragment. The size of the gene was approximately 3.3 kilobases, as expected for a protein with a molecular weight of 104,000.

• Research Organization: Univ. of Illinois, Urbana
• OSTI ID: 5448903
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 164:2; ISSN JOBAA
• Country of Publication: United States
• Language: English

Related Subjects

550400 -- Genetics
550700* -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
AMINES
BACTERIA
CARBOHYDRATES
CHONDROITIN
CLONING
DNA-CLONING
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
ESCHERICHIA COLI
LYASES
MICROORGANISMS
MOLECULAR WEIGHT
MUCOPOLYSACCHARIDES
ORGANIC COMPOUNDS
POLYSACCHARIDES
SACCHARIDES