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Characterization and application of recombinant β-glucosidase (BglH) from Bacillus licheniformis KCTC 1918

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β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC. catalyzes the hydrolysis of β-glucosidic bonds between saccharides and aryl or alkyl groups. A gene encoding β-glucosidase from Bacillus licheniformis KCTC 1918, an anaerobic spore-forming soil bacterium, was cloned and characterized. The structural gene for the β-glucosidase consists of 1410 bp encoding 469 amino acid residues, and has a molecular weight of 53.4 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis with 12% separating gel. The enzyme activity was determined against pNPG as a substrate. The enzyme was optimally active at pH 6.0 (citrate-phosphate buffer) and 47°C. β-Glucosidase retained 100% of its original activity for 24 h. The activity of the enzyme was stimulated by glycerol and urea and was decreased by Ca2+, Cu2+, Hg2+, Mg2+, and Mn2+. In particular, Cu2+ had the strongest negative effect on β-glucosidase activity. The purified β-glucosidase was active against pNPG and cellobiose. When the β-glucosidase was tested for cellulose hydrolysis, the supplement of β-glucosidase with cellulose increased the glucose yield from pine wood powder by 139.8%.


  1. 1.

    Caralho AFAC, Goncalves AZ, da Silva R, Gomes E (2006) A specific short dextrin-hydrolyzing extracellular glucosidase from the thermophilic fungus Thermoascus aurantiacus 179-5. J Microbiol 44:276–283

  2. 2.

    Goyal K, Selvakumar P, Hayashi K (2001) Characterization of a thermostable β-glucosidase (BglB) from Thermotoga maritima showing transglycosylation activity. J Mol Catal B 15:45–53

  3. 3.

    Kim SJ, Lee CM, Kim MY, Yeo YS, Yoon SH, Kang HC, Koo BS (2007) Screening and characterization of an enzyme with β-glucosidase activity from environmental DNA. J Microbiol Biotechnol 17:905–912

  4. 4.

    Park JN, Kim HO, Shin DJ, Kim HJ, Lee HB, Chun SB, Bai S (2001) Cloning of a Paenibacillus sp. endo-β-1,4-glucanase gene and its coexpression with the Endomyces fibuliger β-glucosidase gene in Saccharomyces cerevisiae. J Microbiol Biotechnol 11: 685–692

  5. 5.

    Yoshida M, Igarashi K, Kawai R, Aida K, Samejima M (2004) Differential transcription of β-glucosidase and cellobiose dehydrogenase genes in cellulose degradation by the basidiomycete Phanerochaete chrysosporium. FEMS Microbiol Lett 235:177–182

  6. 6.

    Maheshwari R, Bharadwaj G, Mahalingeshwara KB (2000) Thermophilic fungi: their physiology and enzymes. Microbiol Mol Biol Rev 64:461–488

  7. 7.

    Bhat MK, Bhat S (1997) Cellulose degradation enzymes and their potential industrial applications. Biotechnol Adv 15:583.620

  8. 8.

    Harhangi HR, Steenbakkers P, Akhmanova AS, Jetten MSM, van der Drift C, Op den Camp HJM (2002) A highly expressed family 1 β-glucosidase with transglycosylation capacity from the anaerobic fungus Piromyces sp. E2. Biochim Biophys Acta Gene Struct Expr 1574:293–303

  9. 9.

    Paavilainen S, Hellman J, Korpela T (1993) Purification, characterization, gene cloning, and sequencing of a new β-glucosidase from Bacillus circulans subsp. alkalophilus. Appl Environ Microb 59:927–932

  10. 10.

    Henrissat B (1991) A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 280:309.316

  11. 11.

    Henrissat B, Davies G (1997) Structural and sequence based classification of glycoside hydrolases. Curr Opin Struct Biol 7:637–644

  12. 12.

    Holm L, Sander C (1994) Structural similarity of plant chitinase and lysozymes from animals and phage. FEBS Lett 340:129–132

  13. 13.

    Singh A, Hayashi K (1995) Construction of chimeric β-glucosidases with improved enzymatic properties. J Biol Chem 270:21928–21933

  14. 14.

    Painbeni E, Valles S, Polaina J, Flors A (1992) Purification and characterization of a Bacillus polymyxa β-glucosidase expressed in Escherichia coli. J Bacteriol 174:3087–3091

  15. 15.

    Kuo LC, Lee KT (2008) Cloning, expression, and characterization of two β-glucosidases from isoflavone glycosidase-hydrolyzing Bacillus subtilis natto. J Agr Food Chem 56:119–125

  16. 16.

    Tobisch S, Glaser P, Kruger S, Hecker M (1997) Identification and characterization of a new β-glucoside utilization system in Bacillus subtilis. J Bacteriol 179:496–506

  17. 17.

    Bollet C, Gevaudan MJ, Lamballerie X, Zandotti C, Micco P (1991) A simple method for the isolation of chromosomal DNA from gram positive or acid-fast bacteria. Nucleic Acids Res 19: 1955

  18. 18.

    Kwon KS, Kang HG, Hah YC (1992) Purification and characterization of two extracellular β-glucosidases from Aspergillus nidulans. FEMS Microbiol Lett 97:149–154

  19. 19.

    Bae HJ, Turcotte G, Chamberland H, Karita S, Vezina LP (2003) A comparative study between an endoglucanase IV and its fused protein complex Cel5-CBM6, FEMS Microbiol Lett 227:175–181

  20. 20.

    Archana A, Satyanarayana T (2003) Purification and characterization of a cellulase-free xylanase of a moderate thermophile Bacillus licheniformis A99. World J Microb Biotechnol 19:53–57

  21. 21.

    Abel M, Iversen K, Planas A, Christensen U (2003) Presteady-state kinetics of Bacillus licheniformis 1,3-1,4-β-glucanase: evidence for a regulatory binding state. Biochem J 371:997–1003

  22. 22.

    Igarashi K, Tani T, Kawaki R, Samegima M (2003) Family 3 β-glucosidase from cellulose-degrading culture of the white-rot fungus Phanerochaete chrysosporium is a glucan 1,3-β-glucosidase. J Biosci Bioeng 95:572–576

  23. 23.

    Karnchanatat A, Petsom A, Sanvanich P, Piaphukiew J, Whalley AJ, Reynolds CD, Sihanonth P (2007) Purification and biochemical characterization of an extracellular β-glucosidase from the wood-decaying fungus Daldinia eschscholzii (Ehrenb.:Fr.) Rehm. FEMS Microbiol Lett 270:162–170

  24. 24.

    Rouvinen J, Bergfors T, Teeri T, Knowles J, Jone T (1990) Three-dimensional structure of cellobiohydrolase II from Tricoderma reesei. Science 249:380–386

  25. 25.

    Ohmiya Y, Takeda T, Nakamura S, Sakai F, Hayashi T (1995) Purification and properties of a wall-bound endo-1,4-β-glucanase from suspension-cultured poplar cells. Plant Cell Physiol 36: 607–614

  26. 26.

    Gueguen Y, Chemardin P, Arnaud A, Galzy P (1995) Purification and characterization of an intracellular β-glucosidase from Botrytis cinerea. Enzyme Microb Tech 17:900–906

  27. 27.

    Riou C, Salmon J, Vallier M, Gunata Z, Barre P (1998) Purification, characterization, and substrate specificity of a novel highly glucose tolerant β-glucosidase from Asperillus oryzae. Appl Environ Microb 64:3607–3614

  28. 28.

    Eric P, Salvador V, Julio P, Agusti F (1992) Purification and characterization of a Bacillus polymyxa β-glucosidase expressed in Escherichia coli. J Bacteriol 174:3087–3091

  29. 29.

    Mawadza C, Kaul R, Zvauya R, Mattiasson B (2000) Purification and characterization of cellulase produced by two Bacillus strains. J Biotechnol 83:177–187

  30. 30.

    Yazdi MT, Khosravi AA, Nemati M, Motlagh DV (2003) Purification and characterization of two intracellular β-glucosidases from the Neurospora crassa mutant cell-1. World J Microb Biotechnol 19:79–84

  31. 31.

    Berlin A, Gilkes N, Kilburn D, Bura R, Markov A, Skomarovsky A, Okunev O, Gusakov A, Maximenko V, Gregg D, Sinitsyn A, Saddler J (2005) Evaluation of novel fungal cellulase preparations for ability to hydrolyze softwood substrates — evidence for the role of accessory enzymes. Enzyme Microb Tech 37:175–184

  32. 32.

    Berlin A, Maximenko A, Gilkes N, Saddler J (2007) Optimization of enzyme complexes for lignocelluloses hydrolysis. Biotechnol Bioeng 97:287–296

  33. 33.

    Rahman MD, Katayama T, Suzuki T, Yoshihara Y, Nakagawa T (2007) Stereochemistry and biosynthesis of (+)-lyoniresinol, a syringyl tetrahydronaphthalene lignin in Lyonia ovalifolia var. elliptica II: feeding experiments with 14C labeled precursors. J Wood Sci 53:114–120

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Correspondence to Hyeun-Jong Bae.

Additional information

Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007

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Choi, I.S., Wi, S.G., Jung, S.R. et al. Characterization and application of recombinant β-glucosidase (BglH) from Bacillus licheniformis KCTC 1918. J Wood Sci 55, 329–334 (2009) doi:10.1007/s10086-009-1044-2

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Key words

  • Cellulose
  • Cellulose degradation
  • β-Glucosidase