1887

Abstract

The gene for thermostable 1,3-β-glucosidase BgIB was cloned from the chromosome of and its primary sequence was determined. The purified recombinant β-glucosidase B had a monomer molecular mass of 81 kDa in accordance with the amino acid sequence predicted from the nucleotide sequence of clone pTT51. It was a member of glycosylhydrolase family 3 and belonged to enzyme class EC 3.2.1.21. β-Glucosidase B had a specific activity of 255 U mgon 4-nitrophenyl(PNP)-β-glucoside at the optima of pH (5.5) and temperature (90 °C), and values of 0.1, 10 and 50 mM for PNP-β-glucoside, laminaribiose and cellobiose, respectively. The gene was located immediately upstream of the laminarinase gene Both genes were transcribed from the same DNA strand and were not separated by a palindromic transcription terminator. The two purified enzymes 1,3-β-glucosidase BgIB (laminaribiase) and 1,3-β-glucanase LamA (laminarinase) were together capable of completely degrading laminarin to glucose.

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1997-11-01
2021-07-29
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References

  1. Bause E., Legler G. 1980; Isolation and structure of a tryptic glycopeptide from the active site of β-glucosidase A3 from Aspergillus wentii. . Biochim Biophys Acta 626:459–465
    [Google Scholar]
  2. Bok J.D., Goers S.K., Eveleigh D.E. 1994; Cellulase and xylanase systems of Thermotoga neapolitana. . In Enzymatic Conversion of Biomass for Fuel Production pp. 54–65 Edited by Himmel M. E., Baker J. O., Overend R. P. ACS Symposium series 566. Washington, DC: American Chemical Society;
    [Google Scholar]
  3. Dakhova O.N., Kurepina N.E., Zverlov V.V., Svetlichnyi V.A., Velikodvorskaya G.A. 1993; Cloning and expression in Escherichia coli of Thermotoga neapolitana genes coding for enzymes of carbohydrate substrate degradation.. Biochem Biophys Res Commun 194:1359–1364
    [Google Scholar]
  4. Gräbnitz F., Rücknagel K.P., SeiB M., Staudenbauer W.L. 1989; Nucleotide sequence of the Clostridium thermocellum bglB gene encoding thermostable β-glucosidase B: homology to fungal β-glucosidases.. Mol Gen Genet 217:70–76
    [Google Scholar]
  5. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids.. J Mol Biol 16:557–580
    [Google Scholar]
  6. Henrissat B., Bairoch A. 1993; New families in the classification of glycosyl hydrolases based on amino acid sequence similarities.. Biochem J 280:309–316
    [Google Scholar]
  7. Huber R., Stetter K.O. 1992; The order Thermotogales. . In The Prokaryotes, 2nd edn. pp. 3809–3815 Edited by Balows A., Trüper H. G., Dworkin M., Harder W., Schleifer K.-H. New York: Springer;
    [Google Scholar]
  8. Liebl W., Gabelsberger J., Schleifer K.-H. 1994; 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-l,4-glycosyl hydrolases.. Mol Gen Genet 242:111–115
    [Google Scholar]
  9. Podbielski A., Pohl B., Woischnik M., Körner C., Schmidt K.-H., Rozdzinski E., Leonard B.A.B. 1996; Molecular characterization of group A streptococcal (GAS) oligopeptide permease (Opp) and its effect on cystein protease production.. Mol Microbiol 21:1087–1099
    [Google Scholar]
  10. Rojas A., Romeu A. 1996; A sequence analysis of the β- glucosidase sub-family B.. FEBS Lett 378:93–97
    [Google Scholar]
  11. Ruttersmith L.D., Daniel R.M. 1993; Thermostable β- glucosidase and β-xylosidase from Thermotoga sp. Strain FjSS3- B.l.. Biochim Biophys Acta 1156:167–172
    [Google Scholar]
  12. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  13. Wood T.M., Bhat K.M. 1988; Methods for measuring cellulase activities.. Methods Enzymol 160:87–112
    [Google Scholar]
  14. Zverlov V., Piotukh K., Dakhova O., Velikodvorskaya G., Borriss R. 1996; The multidomain xylanase A of the hyper-thermophilic bacterium Thermotoga neapolitana is extremely thermoresistant.. Appl Microbiol Biotechnol 45:245–247
    [Google Scholar]
  15. Zverlov V.V., Volkov I.Y., Velikodvorskaya T.V., Schwarz W.H. 1997; Highly thermostable endo-l,3-β-glucanase (laminar- inase) LamA from Thermotoga neapolitana: nucleotide sequence of the gene and characterization of the recombinant gene product.. Microbiology 143:1701–1708
    [Google Scholar]
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