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Applied and Environmental Microbiology, November 1998, p. 4217-4225, Vol. 64, No. 11
Department of
Genetics1 and
Department of Biochemistry
and Molecular Biology,2 University of
Georgia, Athens, Georgia
Received 16 June 1998/Accepted 25 August 1998
Xylanase activity assays were used to screen a Streptomyces
coelicolor genomic library in Escherichia coli, and a
xylanase gene that is 99% identical to the xylanase B gene
(xlnB) of S. lividans (GenBank accession
no. M64552) was identified. The promoter region of this gene was
identified by using a transcriptional fusion between the upstream
region of the S. coelicolor xlnB gene and the
xylE reporter gene. Transcription from the xlnB
promoter was found to be induced by xylan and repressed by glucose. A
single apparent transcription start site was identified by both primer extension analysis and in vitro run off transcription assays. Analysis
of deletions of the promoter identified a region required for glucose
repression. By using the transcriptional and protein localization
signals of the Streptomyces xlnB gene, an in-frame translational fusion between the end of the xlnB signal
sequence and the ATG of the Thermotoga maritima xynA gene
was constructed. The xynA gene encodes a thermostable
xylanase that has been demonstrated to be useful in the bleaching of
Kraft pulp. The xlnB-xynA gene fusion was expressed in
Streptomyces, and the activity of the protein produced was
thermostable and was localized to the supernatant fraction of harvested cells.
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Partial Characterization of the Streptomyces lividans
xlnB Promoter and Its Use for Expression of a
Thermostable Xylanase from Thermotoga maritima
*
Corresponding author. Mailing address: Department of
Genetics, Life Sciences Building, University of Georgia, Athens, GA
30602-7223. Phone: (706) 542-1436. Fax: (706) 542-3910. E-mail:
JANWEST{at}ARCHES.UGA.EDU.
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