Abstract
The primary structure of the bglA gene region encoding a β-glucosidase of Thermotoga maritima strain MSB8 was determined. The bglA gene has the potential to code for a polypeptide of 446 amino acids with a predicted molecular mass of 51545 Da. The T, maritima β-glucosidase (BglA) was overexpressed in E. coli at a level comprising approximately 15–20% of soluble cellular protein. Based on its amino acid sequence, as deduced from the nucleotide sequence of the gene, BglA can be classified as a broad-specificity β-glucosidase and as a member of the β-glucosidase family BGA, in agreement with the results of enzymatic characterization of the recombinant protein. Comparative sequence analysis revealed distant amino acid sequence similarities between BGA family β-glucosidases, a β-xylosidase, β-1,4-glycanases of the enzyme family F (mostly xylanases), and other families of β-1,4-glycosyl hydrolases. This result indicates that BGA β-glucosidases may comprise one enzyme family within a large ‘enzyme order’ of retaining β-glycosyl hydrolases, and that the members of these enzyme groups may be inter-related at the level of active site architecture and perhaps even on the level of overall three-dimensional fold.
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References
Ait N, Cruezet N, Cattaneo J (1982) Properties of β-glucosidase purified from Clostridium thermocellum. J Gen Microbiol 128:569–577
Andersson SGE, Kurland CG (1990) Codon preferences in freeliving microorganisms. Microbiol Rev 54:98–210
Baird SD, Hefford MA, Johnson DA, Sung WL, Yaguchi M, Segily VL (1990) The Glu residue in the conserved Asn-Glu-Pro sequence of two highly divergent endo-β-1,4-glucanases is essential for enzymatic activity. Biochem Biophys Res Commun 169:1035–1039
Beguin P (1990) Molecular biology of cellulose degradation. Annu Rev Microbiol 44:219–248
Day AG, Withers SG (1986) Purification and characterization of a β-glucosidase from Alcaligenes faecalis. Can J Biochem Cell Biol 64:914–922
Gabelsberger J, Liebl W, Schleifer KH (1993a) Cloning and characterization of β-galactoside and β-glucoside hydrolysing enzymes of Thermotoga maritima. FEMS Microbiol Lett 109:131–138
Gabelsberger J, Liebl W, Schleifer KH (1993b) Purification and properties of recombinant β-glucosidase of the hyperthermophilic bacterium Thermotoga maritima. Appl. Microbiol. Biotechnol, in press
Gebler JC, Aebersold R, Withers SG (1992a) Glu-537, not Glu-461, is the nucleophile in the active site of (lacZ) β-galactosidase from Escherichia coli. J Biol Chem 267:11126–11130
Gebler J, Gilkes NR, Claeyssens M, Wilson DB, Beguin P, Wakarchuk WW, Kilburn DG, Miller RC Jr, Warren RAJ, Withers SG (1992b) Stereoselective hydrolysis catalyzed by related β-1,4-glucanases and β-1,4-xylanases. J Biol Chem 267:12559–12561
Gilkes NR, Langsford ML, Kilburn DG, Miller RC Jr, Warren RAJ (1984) Mode of action and substrate specificities of cellulases of cloned bacterial genes. J Biol Chem 259:10455–10459
Gräbnitz F, Seiss M, Rücknagel KP, Staudenbauer WL (1991) Structure of the β-glucosidase gene bg1A of Clostridium thermocellum. Eur J Biochem 200:301–309
Henrissat B (1991) A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 280:309–316
Lüthi E, Love DR, McAnulty J, Wallace C, Caughey PA, Saul D, Bergquist PL (1990) Cloning, sequence analysis, and expression of genes encoding xylan-degrading enzymes from the thermophile Caldocellum saccharolyticum. Appl Environ Microbiol 56:1017–1024
Patchett ML, Daniel RM, Morgan HW (1987) Purification and properties of a stable β-glucosidase from an extremely thermophilic anaerobic bacterium. Biochem J 243:779–787
Platt T (1986) Transcription termination and regulation of gene expression. Ann Rev Biochem 55:339–372
Py B, Bortoli-German I, Haiech J, Chippaux M, Barras F (1991) Cellulase EGZ of Erwinia chrysanthemi: structural organization and importance of His 98 and importance of His 98 and Glu 133 residue for catalysis. Prot Eng 4:325–333
Sanger F, Micklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Trimbur DE, Warren RAJ, Withers SG (1992) Region-directed mutagenesis of residues surrounding the active site nucleophile in β-glucosidase from Agrobacterium faecalis. J Biol Chem 267:10248–10251
Tull D, Withers SG, Gilkes NR, Kilburn DG, Warren RAJ, Aebersold R (1991) Glutamic acid 274 is the nucleophile in the active site of a “retaining” exoglucanase from Cellulomonas fimi. J Biol Chem 266:15621–15625
Withers SG, Warren RAJ, Street IP, Rupitz K, Kempton JB, Aebersold R (1990) Unequivocal demonstration of a glutamate residue as a nucleophile in the mechanism of a “retaining” glucosidase. J Am Chem Soc 112:5887–5889
Wright RM, Yablonsky MD, Shalita ZP, Goyal AK, Eveleigh DE (1992) Cloning, characterization and nucleotide sequence of a gene encoding Microbispora bispora BglB, a thermostable β-glucosidase expressed in Escherichia coli. Appl Environ Microbiol 58:3455–3465
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Liebl, W., Gabelsberger, J. & Schleifer, KH. Comparative amino acid sequence analysis of Thermotoga maritima β-glucosidase (BglA) deduced from the nucleotide sequence of the gene indicates distant relationship between β-glucosidases of the BGA family and other families of β-1,4-glycosyl hydrolases. Molec. Gen. Genet. 242, 111–115 (1994). https://doi.org/10.1007/BF00277355
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DOI: https://doi.org/10.1007/BF00277355