Molecular cloning and expression of a xylanase gene from Bacillus polymyxa in Escherichia coli

Journal Article · Fri Apr 01 04:00:00 EST 1988 · Appl. Environ. Microbiol.; (United States)

OSTI ID:6958327

Yang, R C.A.; MacKenzie, R; Bilous, D

Genomic fragments of Bacillus polymyxa derived form separate and complete digestion by EcoRI, HindIII, and BamHI were ligated into the corresponding sites of pBR322, and the resulting chimeric plasmids were transformed into Escherichia coli. Of 6000 transformants screened, 1 (pBPX-277) produced a clear halo on Remazol brilliant blue xylan plates. The insert in the pBPX-277 recombinant, identified as an 8.0-kilobase BamHI fragment of B. polymyxa, was subsequently subjected to extensive mapping and a series of subclonings into pUC19. A 2.9-kilobase BamHI-EcoRI subfragment was found to code for xylanase activity. Xylanase activity expressed by E. coli harboring the cloned gene was located primarily in the periplasm and corresponded to one of two distinct xylanases produced by B. polymyxa. Xylanase expression by the cloned gene occurred in the absence of xylan and was reduced by glucose and xylose. Southern blot hybridization with the cloned fragment as a probe against complete genomic digests of the bacilli B. polymyxa, B. circulans, and B. subtilis revealed that the cloned xylanase gene was unique to B. polymyxa. The xylanase expressed by the cloned gene had a molecular weight of approximately 48,000 and an isoelectric point of 4.9.

• Research Organization: National Research Council of Canada, Ottawa
• OSTI ID: 6958327
• Journal Information: Appl. Environ. Microbiol.; (United States), Journal Name: Appl. Environ. Microbiol.; (United States) Vol. 54:4; ISSN AEMID
• Country of Publication: United States
• Language: English

Related Subjects

500200* -- Environment
Atmospheric-- Chemicals Monitoring & Transport-- (-1989)
54 ENVIRONMENTAL SCIENCES
BACILLUS
BACTERIA
CLONING
DNA
DNA-CLONING
ENZYMES
ESCHERICHIA COLI
GENE REGULATION
GENETIC ENGINEERING
GENETIC MAPPING
GLYCOSYL HYDROLASES
HYBRIDIZATION
HYDROLASES
MAPPING
MICROORGANISMS
NUCLEIC ACIDS
O-GLYCOSYL HYDROLASES
ORGANIC COMPOUNDS
RECOMBINANT DNA
XYLANASE