Cloning and expression of the malolactic gene of Pediococcus damnosus NCFB1832 in Saccharomyces cerevisiae

• Published in: Journal of biotechnology 2005-09, Vol.118 (4), p.353-362
• Main Authors: Bauer, Rolene, Volschenk, Heinrich, Dicks, Leon M.T.
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - genetics; Base Sequence; Biological and medical sciences; Biotechnology; Cloning; Cloning, Molecular; Fundamental and applied biological sciences. Psychology; Gene Expression; gene transfer; genes; grape must; lactic acid bacteria; Lactococcus lactis; Lactococcus lactis - enzymology; Lactococcus lactis - genetics; Lactococcus lactis subsp. lactis; Malate Dehydrogenase - genetics; Malates - metabolism; malic acid; Malolactic fermentation; mleA gene; mleD gene; mleS gene; Molecular Sequence Data; nucleotide sequences; Oenococcus oeni; Pediococcus; Pediococcus - enzymology; Pediococcus - genetics; Pediococcus damnosus; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Schizosaccharomyces pombe; Vitaceae; Wine - microbiology; winemaking
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2005.04.015

Wine production is characterized by a primary alcoholic fermentation, conducted by Saccharomyces cerevisiae, followed by a secondary malolactic fermentation (MLF). Although most lactic acid bacteria (LAB) have the ability to metabolize l-malate, only a few species survive the high ethanol and SO 2 levels in wine. Wines produced in colder viticultural regions have a lower pH than wines produced in warmer regions. The decarboxylation of l-malate in these wines leads to an increase in pH, more organoleptic complexity and microbiological stability. MLF is, however, difficult to control and problems often occur during filtering of such wines. Pediococcus spp. are known to occur in high pH wines and have strong malolactic activity. However, some pediococci synthesize exocellular polysaccharides, which may lead to abnormal viscosity in wine. In this study, the malolactic gene from Pediococcus damnosus NCFB1832 ( mleD) was cloned into S. cerevisiae and co-expressed with the malate permease gene ( mae1) of Schizosaccharomyces pombe. Expression of the mleD gene was compared to the expression of two other malolactic genes, mleS from Lactococcus lactis MG1363 and mleA from Oenococcus oeni Lal1. The genetically modified strain of S. cerevisiae decreased the level of l-malate in grape must to less than 0.3 g l −1 within 3 days. This is the first expression of a malolactic gene from Pediococcus in S. cerevisiae.