Functional expression in Saccharomyces cerevisiae of the Lactococcus lactis mleS gene encoding the malolactic enzyme

Denayrolles, Muriel

doi:10.1016/0378-1097(94)00489-e

Cited by 8 publications

(10 citation statements)

References 6 publications

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“…Genetic engineering of wine yeast to conduct alcoholic fermentation and malate degradation simultaneously has been explored by several groups. In order to engineer a malolactic pathway in S. cerevisiae the malolactic genes (mleS) from Lactococcus lactis, [2,3,11,28] Lactobacillus delbrueckii [184] and the mleA gene from O. oeni [83] were cloned and expressed in S. cerevisiae. The mleS gene encodes a NAD-dependent malolactic enzyme that converts L-malate to L-lactate and carbon dioxide [28].…”

Section: Bio-adjustment Of Wine Aciditymentioning

confidence: 99%

Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking

Pretorius¹

2000

Yeast

220

346

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Yeasts are predominant in the ancient and complex process of winemaking. In spontaneous fermentations, there is a progressive growth pattern of indigenous yeasts, with the final stages invariably being dominated by the alcohol-tolerant strains of Saccharomyces cerevisiae. This species is universally known as the 'wine yeast' and is widely preferred for initiating wine fermentations. The primary role of wine yeast is to catalyze the rapid, complete and efficient conversion of grape sugars to ethanol, carbon dioxide and other minor, but important, metabolites without the development of off-flavours. However, due to the demanding nature of modern winemaking practices and sophisticated wine markets, there is an ever-growing quest for specialized wine yeast strains possessing a wide range of optimized, improved or novel oenological properties. This review highlights the wealth of untapped indigenous yeasts with oenological potential, the complexity of wine yeasts' genetic features and the genetic techniques often used in strain development. The current status of genetically improved wine yeasts and potential targets for further strain development are outlined. In light of the limited knowledge of industrial wine yeasts' complex genomes and the daunting challenges to comply with strict statutory regulations and consumer demands regarding the future use of genetically modified strains, this review cautions against unrealistic expectations over the short term. However, the staggering potential advantages of improved wine yeasts to both the winemaker and consumer in the third millennium are pointed out.

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Section: Bio-adjustment Of Wine Aciditymentioning

confidence: 99%

Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking

Pretorius¹

2000

Yeast

220

346

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“…No color change was observed with CBT313 carrying pJF119HE. L-Malate uptake was checked by using L-[ 14 VOL. 62,1996 MALOLACTIC GENES FROM LEUCONOSTOC OENOS 1279 lation was observed in the strain carrying pJF119HE (Fig.…”

Section: Cloning Of the Mle Locus From Leuconostoc Oenosmentioning

confidence: 99%

“…A positive regulatory gene called mleR, which is responsible for activation of the malolactic system was described first (38). Recently, cloning, sequencing, and heterologous expression of the MLE gene from Lactococcus lactis (1,13,14) have been described. A sequence alignment of the protein revealed that there are three highly conserved regions that are also present in proteins that belong to the malic enzyme family.…”

mentioning

confidence: 99%

Cloning and characterization of the genes encoding the malolactic enzyme and the malate permease of Leuconostoc oenos

Labarre¹,

Guzzo²,

Cavin³

et al. 1996

Appl Environ Microbiol

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Using degenerated primers from conserved regions of the protein sequences of malic enzymes, we amplified a 324-bp DNA fragment by PCR from Leuconostoc oenos and used this fragment as a probe for screening a Leuconostoc oenos genomic bank. Of the 2,990 clones in the genomic bank examined, 7 with overlapping fragments were isolated by performing colony hybridization experiments. Sequencing 3,453 bp from overlapping fragments revealed two open reading frames that were 1,623 and 942 nucleotides long and were followed by a putative terminator structure. The first deduced protein (molecular weight, 59,118) is very similar (level of similarity, 66%) to the malolactic enzyme of Lactococcus lactis; as in several malic enzymes, highly conserved protein regions are present. The synthesis of a protein with an apparent molecular mass of 60 kDa was highlighted by the results of labelling experiments performed with Escherichia coli minicells. The gene was expressed in E. coli and Saccharomyces cerevisiae and conferred ''malolactic activity'' to these species. The second open reading frame encodes a putative 34,190-Da protein which has the characteristics of a carrier protein and may have 10 membrane-spanning segments organized around a central hydrophilic core. Energydependent L-[ 14 C]malate transport was observed with E. coli dicarboxylic acid transport-deficient mutants carrying the malate permease-expressing vector. Our results suggest that in Leuconostoc oenos the genes that encode the malolactic enzyme and a malate carrier protein are organized in a cluster. MATERIALS AND METHODS Bacterial strains, plasmids, and media. The strains and plasmids used in this study are listed in Table 1. E. coli CBT313 was kindly provided by B. J. Bachmann, Department of Biology, Yale University, New Haven, Conn. E. coli strains were grown in Luria-Bertani broth or agar (6) at 37ЊC. Antibiotics were used at the following concentrations in the selective media: ampicillin, 50 g ⅐ ml Ϫ1 ; and erythromycin, 200 g ⅐ ml Ϫ1. Leuconostoc oenos Lo84.13 was grown at 30ЊC in FT80 medium (pH 5.3) (10) modified by adding meat extract instead of Casamino Acids. S. cerevisiae RH100-4 was grown in YPG medium (35) supplemented with 2% (wt/vol) galactose at 30ЊC with aeration. A selective medium (35) lacking uracil was used to select for transformant yeast strains. S. cerevisiae was transformed by a method in which lithium acetate and polyethylene glycol were used (20).

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“…Strains of Saccharomyces cerevisiae can only metabolize malate to a limited extent (see Radler, 1993 for a review) and strains of Leuconostoc oenos are currently used to deacidify wine. Several attempts have been made to introduce the bacterial malolactic gene into S. cerevisiae (Ansanay et al, 1993;Denayrolles et al, 1995;Williams et al, 1984). However, the recombinant yeast strains were unable to degrade malate effectively to L-lactate.…”

Section: Introductionmentioning

confidence: 99%

The mae1 gene of Schizosaccharomyces pombe encodes a permease for malate and other C₄ dicarboxylic acids

Grobler

Bauer

Subden

et al. 1995

Yeast

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The mae1 gene of the yeast Schizosaccharomyces pombe was identified on the basis of its ability to complement a mutant defective in the transport of malic acid. Analysis of the DNA sequence revealed an open reading frame of 1314 base pairs, encoding a polypeptide of 438 amino acids with a predicted molecular weight of 49 kDa. A hydropathy profile of the predicted amino acid sequence revealed a protein with ten membrane-spanning or associated domains and hydrophilic N- and C- termini. The predicted secondary structure of the protein in similar to models proposed for other integral membrane proteins from both prokaryotes and eukaryotes. The S. pombe mae1 gene encodes a single mRNA of 1.5 kb. The mea1 gene is expressed constitutively and is not subject to catabolite repression as was previously reported for the malate permease systems of Candida utilis and Hansenula anomala. The mae1 gene was mapped 2842 bp 5' to the MFml gene on chromosome I. Transport assays revealed that the mae1 gene encodes a permease involved in the uptake of L-malate, succinate and malonic acid.

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Functional expression in Saccharomyces cerevisiae of the Lactococcus lactis mleS gene encoding the malolactic enzyme

Cited by 8 publications

References 6 publications

Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking

Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking

Cloning and characterization of the genes encoding the malolactic enzyme and the malate permease of Leuconostoc oenos

The mae1 gene of Schizosaccharomyces pombe encodes a permease for malate and other C₄ dicarboxylic acids

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Functional expression in Saccharomyces cerevisiae of the Lactococcus lactis mleS gene encoding the malolactic enzyme

Cited by 8 publications

References 6 publications

Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking

Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking

Cloning and characterization of the genes encoding the malolactic enzyme and the malate permease of Leuconostoc oenos

The mae1 gene of Schizosaccharomyces pombe encodes a permease for malate and other C4 dicarboxylic acids

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The mae1 gene of Schizosaccharomyces pombe encodes a permease for malate and other C₄ dicarboxylic acids