Abstract
A recombinant putative β-galactosidase from Thermoplasma acidophilum was purified as a single 57 kDa band of 82 U mg−1. The molecular mass of the native enzyme was 114 kDa as a dimer. Maximum activity was observed at pH 6.0 and 90°C. The enzyme was unstable below pH 6.0: at pH 6 its half-life at 75°C was 28 days but at pH 4.5 was only 13 h. Catalytic efficiencies decreased as p-nitrophenyl(pNP)-β-d-fucopyranoside (1067) > pNP-β-d-glucopyranoside (381) > pNP-β-d-galactopyranoside (18) > pNP-β-d-mannopyranoside (11 s−1 mM−1), indicating that the enzyme was a β-glycosidase.
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References
Bhat M, Bhat T (1997) Cellulose degradation enzyme and their potential industrial applications. Biotechnol Adv 15:583–620
Bhatia Y, Mishra S, Bisaria VS (2002) Microbial beta-glucosidases: cloning, properties, and applications. Crit Rev Biotechnol 22:375–407
Chuankhayan P, Rimlumduan T, Svasti J, Cairns JR (2007) Hydrolysis of soybean isoflavonoid glycosides by Dalbergia beta-glucosidases. J Agric Food Chem 55:2407–2412
Cobucci-Ponzano B, Moracci M, Di Lauro B, Ciaramella M, D’Avino R, Rossi M (2002) Ionic network at the C-terminus of the beta-glycosidase from the hyperthermophilic archaeon Sulfolobus solfataricus: functional role in the quaternary structure thermal stabilization. Proteins 48:98–106
D’Auria S, Morana A, Febbraio F, Vaccaro C, De Rosa M, Nucci R (1996) Functional and structural properties of the homogeneous beta-glycosidase from the extreme thermoacidophilic archaeon Sulfolobus solfataricus expressed in Saccharomyces cerevisiae. Protein Expr Purif 7:299–308
D’Auria S, Rossi M, Nucci R, Irace G, Bismuto E (1997) Perturbation of conformational dynamics, enzymatic activity, and thermostability of beta-glycosidase from archaeon Sulfolobus solfataricus by pH and sodium dodecyl sulfate detergent. Proteins 27:71–79
D’Auria S, Moracci M, Febbraio F, Tanfani F, Nucci R, Rossi M (1998) Structure-function studies on beta-glycosidase from Sulfolobus solfataricus. Molecular bases of thermostability. Biochimie 80:949–957
Dion M, Fourage L, Hallet JN, Colas B (1999) Cloning and expression of a beta-glycosidase gene from Thermus thermophilus. Sequence and biochemical characterization of the encoded enzyme. Glycoconj J 16:27–37
Gabelsberger J, Liebl W, Schleifer K (1993) Purification and properties of a recombinant beta-glucosides of the hyper thermophilic bacterium Thermotoga maritima. Appl Micobiol Biotechnol 40:44–52
Kengen SW, Luesink EJ, Stams AJ, Zehnder AJ (1993) Purification and characterization of an extremely thermostable beta-glucosidase from the hyperthermophilic archaeon Pyrococcus furiosus. Eur J Biochem 213:305–312
Leite RS, Gomes E, da Silva R (2007) Characterization and comparison of thermostability of purified beta-glucosidases from a mesophilic Aureobasidium pullulans and a thermophilic Thermoascus aurantiacus. Process Biochem 42:1101–1106
Matsui I, Sakai Y, Matsui E, Kikuchi H, Kawarabayasi Y, Honda K (2000) Novel substrate specificity of a membrane-bound beta-glycosidase from the hyperthermophilic archaeon Pyrococcus horikoshii. FEBS Lett 467:195–200
Petzelbauer I, Nidetzky B, Haltrich D, Kulbe KD (1999) Development of an ultra-high-temperature process for the enzymatic hydrolysis of lactose. I. The properties of two thermostable beta-glycosidases. Biotechnol Bioeng 64:322–332
Takase M, Horikoshi K (1988) A thermostable beta-glucosidase isolated from a bacterial species of the genus Thermus. Appl Microbiol Biotechnol 29:55–60
Venturi LL, Polizeli Mde L, Terenzi HF, Furriel Rdos P, Jorge JA (2002) Extracellular beta-d-glucosidase from Chaetomium thermophilum var. coprophilum: production, purification and some biochemical properties. J Basic Microbiol 42:55–66
Zverlov VV, Volkov IY, Velikodvorskaya TV, Schwarz WH (1997) Thermotoga neapolitana bglB gene, upstream of lamA, encodes a highly thermostable beta-glucosidase that is a laminaribiase. Microbiology 143:3537–3542
Acknowledgments
This study was carried out with the support of ‘Forest Science and Technology Projects (Project No. S210707L010120)’ provided by Korea Forest Service, by the 21C Frontier Project for Microbial Genomics, Ministry of Science and Technology, and by the Korea Research Foundation Grant (MOEHRD) (KRF-2006-351-D00012).
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Kim, HJ., Park, AR., Lee, JK. et al. Characterization of an acid-labile, thermostable β-glycosidase from Thermoplasma acidophilum . Biotechnol Lett 31, 1457–1462 (2009). https://doi.org/10.1007/s10529-009-0018-1
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DOI: https://doi.org/10.1007/s10529-009-0018-1