- Original Article
- Published:
Characterization and application of recombinant β-glucosidase (BglH) from Bacillus licheniformis KCTC 1918
Journal of Wood Science volume 55, pages 329–334 (2009)
Abstract
β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC.3.2.1.21) 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%.
References
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
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
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
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
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
Maheshwari R, Bharadwaj G, Mahalingeshwara KB (2000) Thermophilic fungi: their physiology and enzymes. Microbiol Mol Biol Rev 64:461–488
Bhat MK, Bhat S (1997) Cellulose degradation enzymes and their potential industrial applications. Biotechnol Adv 15:583.620
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
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
Henrissat B (1991) A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 280:309.316
Henrissat B, Davies G (1997) Structural and sequence based classification of glycoside hydrolases. Curr Opin Struct Biol 7:637–644
Holm L, Sander C (1994) Structural similarity of plant chitinase and lysozymes from animals and phage. FEBS Lett 340:129–132
Singh A, Hayashi K (1995) Construction of chimeric β-glucosidases with improved enzymatic properties. J Biol Chem 270:21928–21933
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
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
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
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
Kwon KS, Kang HG, Hah YC (1992) Purification and characterization of two extracellular β-glucosidases from Aspergillus nidulans. FEMS Microbiol Lett 97:149–154
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
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
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
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
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
Rouvinen J, Bergfors T, Teeri T, Knowles J, Jone T (1990) Three-dimensional structure of cellobiohydrolase II from Tricoderma reesei. Science 249:380–386
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
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
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
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
Mawadza C, Kaul R, Zvauya R, Mattiasson B (2000) Purification and characterization of cellulase produced by two Bacillus strains. J Biotechnol 83:177–187
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
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
Berlin A, Maximenko A, Gilkes N, Saddler J (2007) Optimization of enzyme complexes for lignocelluloses hydrolysis. Biotechnol Bioeng 97:287–296
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
Author information
Authors and Affiliations
Corresponding author
Additional information
Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007
Rights and permissions
About this article
Cite this article
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). https://doi.org/10.1007/s10086-009-1044-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10086-009-1044-2