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Cloning, sequence and expression of the gene encoding the malolactic enzyme from Lactococcus lactis
V Ansanay1, S Dequin, B Blondin
1Laboratoire de Microbiologie et Technologie des Fermentations, Institut des Produits de la Vigne, INRA, Montpellier, France.
FEBS Letters
|October 11, 1993
Summary
Lactic acid bacteria perform malolactic fermentation using malolactic enzyme. Researchers isolated the malolactic enzyme gene (mleS) from Lactococcus lactis, enabling L-malate conversion to L-lactate in bacteria and yeast.
Area of Science:
- Microbiology
- Biochemistry
- Molecular Biology
Background:
- Lactic acid bacteria are known for malolactic fermentation.
- Malolactic fermentation is a secondary fermentation process.
- This process is catalyzed by malolactic enzyme, which requires NAD and Mn(2+).
Purpose of the Study:
- To isolate and characterize the gene encoding malolactic enzyme.
- To understand the genetic basis of malolactic fermentation in Lactococcus lactis.
Main Methods:
- Isolation of the malolactic enzyme gene (mleS) from Lactococcus lactis.
- Sequence analysis of the mleS gene and its predicted protein product.
- Expression of the malolactic gene in bacterial and yeast cells.
Main Results:
- The mleS gene was successfully isolated from Lactococcus lactis.
- The gene comprises a single open reading frame, predicting a 59 kDa protein.
- The predicted amino acid sequence shows homology to other malic enzymes.
- Engineered bacterial and yeast cells demonstrated the ability to convert L-malate to L-lactate.
Conclusions:
- The mleS gene codes for the malolactic enzyme responsible for malolactic fermentation.
- The identified gene can be expressed in heterologous systems, enabling L-malate conversion.
- This research provides a molecular basis for understanding and potentially manipulating malolactic fermentation.