Abstract
A gene coding for the endo-β-1,3-1,4-glucanase of B. circulans ATCC21367 was cloned into Escherichia coli. The cloned enzyme hydrolyzed lichenan or barley β-glucan to produce 3-O-β-cellobiosyl-d-glucose as a main product but was inactive with carboxymethyl cellulose, laminarin and xylan. The enzyme, M r=28 kDa, remained within the cytoplasm of E. coli. A 771 bp open reading frame was in the 2 kb PstI fragment of the recombinant plasmid pLL200K. The deduced protein sequence consists of 257 amino acids and has a putative signal peptide of 26 amino acids. The amino acid sequence of the endo-β-1,3-1,4-glucanase showed 68 and 51% homology to previously reported endo-β-1,3-1,4-glucanases from Bacillus strain N-137 and B. brevis, respectively.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J. Mol. Biol. 215: 403-410.
Borriss R, Buettner K, Maentsaelae P (1990) Structure of the β-1,3-1,4-glucanase gene of Bacillus macerans: homologies to other beta-glucanases. Mol. Gen. Genet. 222: 278-283.
Bueno A, Vazquez de Aldana CR, Correa J, Villa TG, del Rey F (1990a) Synthesis and secretion of a Bacillus circulans WL-12 1,3-1,4-β-D-glucanase in Escherichia coli. J. Bacteriol. 172: 2160-2167.
Bueno A, Vazquez de Aldana CR, Correa J, del Rey F (1990b) Nucleotide sequence of a 1,3-1,4-β-glucanase-encoding gene in Bacillus circulans WL-12. Nucl. Acids Res. 18: 4248.
Chen H, Li XL, Ljungdahl LG (1997) Sequencing of a 1,3-1,4-β-D-glucanase (lichenase) from the anaerobic fungus Orpinomyces strain PC-2: properties of the enzyme expressed in Escherichia coli and evidence that the gene has a bacterial origin. J. Bacteriol. 179: 6028-6034.
Enari TM, Markkanen PH (1975) Microbial β-glucanases in brewing. Proc. Am. Soc. Brew. Chem. 33: 13-17.
Gosalbes MJ, Perez-Gonzalez JA, Gonzalez R, Navarro A (1991) Two β-glucanase genes are clustered in Bacillus polymyxa: molecular cloning, expression, and sequence analysis of genes encoding a xylanase and an endo-β-(1,3)-(1,4)-glucanase. J. Bacteriol. 173: 7705-7710.
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351-359.
Juncosa M, Pons J, Dot T, Querol E, Planas A (1994) Identi-fication of active site carboxylic residues in Bacillus licheniformis 1,3-1,4-β-D-glucan 4-glucanohydrolase by site-directed mutagenesis. J. Biol. Chem. 269: 14530-14535.
Lee DS, Chang HG (1995) Cloning and expression of a β-1,3-glucanase gene from Bacillus circulans KCTC3004 in Escherichia coli. Biotechnol. Lett. 17: 355-360.
Lloberas J, Perez-Pons JA, Querol E (1991) Molecular cloning, expression and nucleotide sequence of the endo-β-1,3-1,4-Dglucanase gene from Bacillus licheniformis. Predictive structural analyses of the encoded polypeptide. Eur. J. Biochem. 197: 337-343.
Louw ME, Reid SJ, Watson TG (1993) Characterization, cloning and sequencing of a thermostable endo-(1,3-1,4)-β-glucanaseencoding gene from an alkalophilic Bacillus brevis. Appl. Microbiol. Biotechnol. 38: 507-513.
Rodriguez RL, Tait RC (1983) Recombinant DNA Techniques, An Introduction. London: Addison-Wesley, pp. 162-163.
Tabernero C, Coll PM, Fernandez-Abalos JM, Perez P, Santamaria RI (1994) Cloning and DNA sequencing of bgaA, a gene encoding an endo-β-1,3-1,4-glucanase, from an alkalophilic Bacillus strain (N137). Appl. Environ. Microbiol. 60: 1213-1220.
von Heijne G (1986) A new method for predicting signal cleavage sites. Nucl. Acids Res. 14: 4683-4690.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kim, JY., Kim, HB. & Lee, DS. Cloning and expression of the Bacillus circulans endo-β-1,3-1,4-glucanase gene (bglBC1) in Escherichia coli . Biotechnology Letters 24, 53–57 (2002). https://doi.org/10.1023/A:1013813401511
Issue Date:
DOI: https://doi.org/10.1023/A:1013813401511