Abstract
In this study, a new α-glucosidase gene from Thermoanaerobacter ethanolicus JW200 was cloned and expressed in Escherichia coli by a novel heat-shock vector pHsh. The recombinant α-glucosidase exhibited its maximum hydrolytic activity at 70°C and pH 5.0∼5.5. With p-nitrophenyl-α-D-glucoside as a substrate and under the optimal condition (70°C, pH 5.5), K m and V max of the enzyme was 1.72 mM and 39 U/mg, respectively. The purified α-glucosidase could hydrolyze oligosaccharides with both α-1,4 and α-1,6 linkages. The enzyme also had strong transglycosylation activity when maltose was used as sugar donor. The transglucosylation products towards maltose are isomaltose, maltotriose, panose, isomaltotriose and tetrasaccharides. The enzyme could convert 400 g/L maltose to oligosaccharides with a conversion rate of 52%, and 83% of the oligosaccharides formed were prebiotic isomaltooligosaccharides (containing isomaltose, panose and isomaltotriose).
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Abbreviations
- pNP:
-
p-nitrophenol
- pNPG:
-
p-nitrophenyl-α-D-glucoside
References
Benson D.A., Karsch-Mizrachi I., Lipman D.J., Ostell J. & Sayers E.W. 2009. GenBank. Nucleic Acids Res. 37: D26–D31.
Chiba S. 1997. Molecular mechanism in α-glucosidase and glucoamylase. Biosci. Biotech. Biochem. 61: 1233–1239.
Chung C.H. 2006. Production of glucooligosaccharides and mannitol from Leuconostoc mesenteroides B-742 fermentation and its separation from byproducts. J. Microbiol. Biotechnol. 16: 325–329.
Crittenden R.G. & Playne M.J. 1996. Production, properties and applications of food-grade oligosaccharides. Trends Food Sci. Technol. 7: 353–361.
Duan K.J., Sheu D.C. & Lin C.T. 1995. Transglucosylation of a fungal α-glucosidase — the enzyme properties and correlation of isomaltooligosaccharide production. Ann. N. Y. Acad. Sci. 750: 325–328.
Ferrer M., Golyshina O.V., Plou F.J., Timmis K.N. & Golyshin P.N. 2005. A novel α-glucosidase from the acidophilic archaeon Ferroplasma acidiphilum strain Y with high transglycosylation activity and an unusual catalytic nucleophile. Biochem. J. 391: 269–276.
Giannesi G.C., Polizeli M.D., Terenzi H.F. & Jorge J.A. 2006. A novel α-glucosidase from Chaetomium thermophilum var. coprophilum that converts maltose into trehalose: purification and partial characterisation of the enzyme. Process Biochem. 41: 1729–1735.
Hasler C.M. 1996. Functional foods: the western perspective. Nutr. Rev. 54: S6–10.
Henrissat B. 1991. A classification of glycosyl hydrolases based on amino acid sequence similarity. Biochem. J. 280: 309–316.
Hung V.S., Hatada Y., Goda S. & Lu J. 2005. α-Glucosidase from a strain of deep-sea Geobacillus: a potential enzyme for the biosynthesis of complex carbohydrates. Appl. Microbiol. Biotechnol. 68: 757–765.
Ichikawa Y., Look G.C., Wong C.H. & Kajimoto T. 1992. Synthesis of oligosaccharides using glycosyltransferases. J. Syn. Org. Chem. Jpn. 50: 441–450.
Kobayashi I., Tokuda M., Hashimoto H., Konda T., Nakano H. & Kitahata S. 2003. Purification and characterization of a new type of α-glucosidase from Paecilomyces lilacinus that has transglucosylation activity to produce α-1,3- and α-1,2-linked oligosaccharides. Biosci. Biotech. Biochem. 67: 29–35.
Lucia F.A., Dolores M., De Segura A.G., Dolores L., Miguel A. & Patricia G.A. 2007. Transformation of maltose into prebiotic isomaltooligosaccharides by a novel α-glucosidase from Xantophyllomyces dendrorhous. Process Biochem. 42: 1530–1536.
Mala S., Dvorakova H., Hrabal R. & Kralova B. 1999. Towards regioselective synthesis of oligosaccharides by use of α-glucosidases with different substrate-specificity. Carbohydr. Res. 322: 209–218.
Murase H., Yamauchi R., Kato K., Kunieda T. & Terao J. 1997. Synthesis of a novel vitamin E derivative, 2-(α-D-glucopyranosyl) methyl-2,5,7,8-tetramethylchroman-6-ol, by α-glucosidase-catalyzed transglycosylation. Lipids 32: 73–78.
Olano-Martin E., Mountzouris K.C., Gibson G.R. & Rastall R.A. 2000. In vitro fermentability of dextran, oligodextran and maltodextrin by human gut bacteria. Br. J. Nutr. 83: 247–255.
Prodanovic R., Milosavic N., Sladic D., Zlatovic M., Bozic B. & Velickovic T.C. 2005. Transglucosylation of hydroquinone catalysed by alpha-glucosidase from baker’s yeast. J. Mol. Catal. B. Enzymatic 35: 142–146.
Remaut E., Stanssens P. & Fiers W. 1981. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene 15: 81–93.
Shao W., Wu H. & Pei J. 2006. A plasmid vector controlled by the σ32 factor of Escherichia coli and its use for the expression of heterologous protein. International Patent; PCT WO2006/002574A1.
Wiegel J., Carriera L.H., Mothershed C.P. & Ljungdahl L.G. 1983. Production of ethanol from biopolymers by anaerobic, thermophilic and extreme thermophilic bacteria. II. Thermoanaerobacter ethanolicus JW200 and its mutants in batch cultures and resting cell experiments. Biotechnol. Bioeng. Symp. 13: 193–205.
Wiegel J. & Ljungdahl L.G. 1981. Thermoanaerobacter ethanolicus gen. nov., spec. nov., a new, extreme thermophilic, anaerobic bacterium. Arch. Microbiol. 128: 343–348.
Wu H., Pei J., Wu G. & Shao W. 2008. Overexpression of GH10 endoxylanase XynB from Thermotoga maritima in Escherichia coli by a novel vector with potential for industrial application. Enzyme. Microb. Technol. 42: 230–234.
Yin E., Le Y., Pei J., Shao W. and Yang Q. 2008. High-level expression of the xylanase from Thermomyces lanuginosus in Escherichia coli. World. J. Microbiol. Biotechnol. 24: 275–280.
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Wang, YH., Jiang, Y., Duan, ZY. et al. Expression and characterization of an α-glucosidase from Thermoanaerobacter ethanolicus JW200 with potential for industrial application. Biologia 64, 1053–1057 (2009). https://doi.org/10.2478/s11756-009-0197-1
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DOI: https://doi.org/10.2478/s11756-009-0197-1