Document Detail

Isolation of the DLD gene of Saccharomyces cerevisiae encoding the mitochondrial enzyme D-lactate ferricytochrome c oxidoreductase.
MedLine Citation:
PMID:  8492799     Owner:  NLM     Status:  MEDLINE    
In Saccharomyces cerevisiae the utilization of lactate occurs via specific oxidation of L- and D-lactate to pyruvate catalysed by L-lactate ferricytochrome c oxidoreductase (L-LCR) (EC encoded by the CYB2 gene, and D-lactate ferricytochrome c oxidoreductase (D-LCR) (EC, respectively. We selected several lactate- pyruvate+ mutants in a cyb2 genetic background. Two of them were devoid of D-LCR activity (dld mutants, belonging to the same complementation group). The mutation mapped in the structural gene. This was demonstrated by a gene dosage effect and by the thermosensitivity of the enzyme activity of thermosensitive revertants. The DLD gene was cloned by complementation for growth on D-, L-lactate in the strain WWF18-3D, carrying both a CYB2 disruption and the dld mutation. The minimal complete complementing sequence was localized by subcloning experiments. From the sequence analysis an open reading frame (ORF) was identified that could encode a polypeptide of 576 amino-acids, corresponding to a calculated molecular weight of 64000 Da. The deduced protein sequence showed significant homology with the previously described microsomal flavoprotein L-gulono-gamma-lactone oxidase isolated from Rattus norvegicus, which catalyses the terminal step of L-ascorbic acid biosynthesis. These results are discussed together with the role of L-LCR and D-LCR in lactate metabolism of S. cerevisiae.
T Lodi; I Ferrero
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular & general genetics : MGG     Volume:  238     ISSN:  0026-8925     ISO Abbreviation:  Mol. Gen. Genet.     Publication Date:  1993 Apr 
Date Detail:
Created Date:  1993-06-17     Completed Date:  1993-06-17     Revised Date:  2010-04-12    
Medline Journal Info:
Nlm Unique ID:  0125036     Medline TA:  Mol Gen Genet     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  315-24     Citation Subset:  IM    
Institute of Genetics, University of Parma, Italy.
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MeSH Terms
Amino Acid Sequence
Base Sequence
Cloning, Molecular
Cytochromes / biosynthesis
Genes, Fungal*
L-Lactate Dehydrogenase / genetics*
Lactate Dehydrogenases*
Mitochondria / enzymology
Molecular Sequence Data
Saccharomyces cerevisiae / enzymology*,  genetics
Reg. No./Substance:
0/Cytochromes; EC 1.1.-/Lactate Dehydrogenases; EC Dehydrogenase; EC dehydrogenase (cytochrome)

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