Abstract
An integrating plasmid pMGI6 carrying glucoamylase gene (GLA) and using the yeast α-acetolactate synthase gene (ILV2) as the recombination sequence, was constructed from pBluescript II SK. Theilv2∶∶GLA fragment released from pMGI6 was introduced into the brewing yeastSaccharomyces pastorianus and the resulting recombinant strain was able to utilise starch as the sole carbon source, its glucoamylase activity was 6.3 U ml−1 and its a-acetolactate synthase activity was lowered by 33%. Fermentation tests confirmed that the diacetyl concentration in wort fermented by the recombinant strain was reduced by 66% and the maturation time was reduced from 7 to 4 days. The beer fermented by the recombinant strain under industrial operating conditions satisfied the high quality demands and the strain could be used in beer production safely.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bamforth C.W. (2002). Nutritional aspects of beer-a review. Nutr. Res., 22: 227–237.
Birol G., Önsan Z.Ï., Klrdar B., Oliver S.G. (1998). Ethanol production and fermentation characteristics of recombinantSaccharomyces cerevsiae strains grown on starch. Enzyme Microb. Technol., 22: 672–677.
Burke D., Dawson D., Stearns T., Eds (2000). Methods in Yeast Genetics, Cold Spring Harbor Laboratory Course Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Cortacero-Ramírez S., Hernáinz-Bermúdez de Castro M., Segura-Carretero A., Cruces-Blanco C., Fernández-Gutiérrez A. (2003). Analysis of beer components by capillary electrophoretic methods. Trends Anal. Chem., 22: 7–8.
Faridmoayer A., Scaman C.H. (2004). An improved purification procedure for soluble processing α-glucosidase I fromSaccharomyces cerevisiae overexpressing CWH41. Protein Expr. Purif., 33: 11–18.
Gundlapalli Moses S.B., Cordero Otero R.R., La Grange D.C., van Rensburg, P., and Pretorius, I.S. (2002). Different genetic backgrounds influence the secretory expression of theLKA1-encoddedLipomyces kononenkoae α-amylase in industrial strains ofSaccharomyces cerevisiae. Biotechnol. Lett., 24: 651–656.
Hill J.E., Meyers A.M., Koerner T.J., Tzagoloff A. (1993). Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast, 9: 163–167.
Kang N.Y., Park J.N., Chin J.E., Lee H.B., Im S.Y., Bai S. (2003). Construction of an amylolytic industrial strain ofSaccharomyces cerevisiae containing theSchwanniomyces occidentalis α-amylase gene. Biotechnol. Lett., 25: 1847–1851.
Liu Z.R., Zhang G.Y., Liu S.G. (2004). Constructing an amylolytic brewing yeastSaccharomyces pastorianus suitable for accelerated brewing. J. Biosci. Bioeng., 98: 414–419.
Nevoigt E.; Pilger R.; Mast-Gerlach E.; Schmidt U.; Freihammer S.; Eschenbrenner M., Garbe L., Stahl U. (2002). Genetic engineering of brewing yeast to reduce the content of ethanol in beer. FEMS Yeast Res., 2: 225–232.
Park, S.H., Xing, R., Whitman, W.B. (1995). Nonenzymatic acetolactate oxidation to diacetyl by flavin, nicotinamide and quinine coenzymes. Biochem. Biophys. Acta, 1245: 366–370.
Parent S.A., Fenimore C.M., Bostian K.A. (1985). Vector systems for the expression, analysis and cloning of DNA sequences inS. cerevisiae. Yeast, 1: 83–138.
Sambrook J., Russell D.W., Eds (2001). Molecular Cloning, A Laboratory Manual, 3rd edn., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Shigechi H., Koh J., Fujita Y., Matsumoto T., Bito Y., Ueda M., Satoh E., Fukuda H., Kondo A. (2004). Direct production of ethanol from raw corn starch via fermentation by use of a novel surface-engineered yeast strain co-displaying glucoamylase and α-amylase. Appl. Environ. Microbiol., 70: 5037–5040.
Short J.M., Fernandez J.M., Sorge J.A., and Huse W.D. (1988). ψ ZAP: A bacteriophage ψ expression vector with in vivo excision properties. Nucleic Acids Res., 16: 7583.
Steyn A.J.C., Pretorius I.S. (1991). Co-expression of aSaccharomyces diastaticus glucoamylase-encoding gene and aBacillus amyloliquefaciens α-amylase-encoding gene inSaccharomyces cerevisiae. Gene, 100: 85–93.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Liu, ZR., Zhang, GY., Li, J. et al. Stable expression of glucoamylase gene in industrial strain ofSaccharomyces pastorianus with less diacetyl produced. Ann. Microbiol. 57, 233–237 (2007). https://doi.org/10.1007/BF03175212
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF03175212