Identification and characterization of a novel glucose-phosphorylating enzyme in Kluyveromyces lactis

Recent data suggest that hexokinase KlHxk1 (Rag5) represents the only glucose-phosphorylating enzyme of Kluyveromyces lactis, which also is required for glucose signalling. Long-term growth studies of a K. lactis rag5 mutant, however, reveal slow growth on glucose, but no growth on fructose. Isolati...

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Bibliographic Details
Published in:FEMS yeast research 2007-08, Vol.7 (5), p.683-692
Main Authors: Kettner, Karina, Müller, Eva-Christina, Otto, Albrecht, Rödel, Gerhard, Breunig, Karin D, Kriegel, Thomas M
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Enzymes
Fructose
Fructose - metabolism
Fungal Proteins - chemistry
Fungal Proteins - genetics
Fungal Proteins - isolation & purification
Fungal Proteins - metabolism
Gene Expression Regulation, Fungal
Genomes
Glucokinase
Glucokinase - chemistry
Glucokinase - genetics
Glucokinase - isolation & purification
Glucokinase - metabolism
Glucose
Glucose - metabolism
Hexokinase
Humans
KlGlk1
KlHxk1
Kluyveromyces - enzymology
Kluyveromyces - genetics
Kluyveromyces - growth & development
Kluyveromyces lactis
Molecular Sequence Data
Mutants
Nucleotide sequence
Open reading frames
Phosphorylation
Phylogeny
Sequence Alignment
Substrate Specificity
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Summary:Recent data suggest that hexokinase KlHxk1 (Rag5) represents the only glucose-phosphorylating enzyme of Kluyveromyces lactis, which also is required for glucose signalling. Long-term growth studies of a K. lactis rag5 mutant, however, reveal slow growth on glucose, but no growth on fructose. Isolation of the permissive glucose-phosphorylating enzyme, mass spectrometric tryptic peptide analysis and determination of basic kinetic data identify a novel glucokinase (KlGlk1) encoded by ORF KLLA0C01155g. In accordance with the growth characteristics of the rag5 mutant, KlGlk1 phosphorylates glucose, but fails to act on fructose as a sugar substrate. Multiple sequence alignment indicates the presence of at least one glucokinase gene in all sequenced yeast genomes.
ISSN:1567-1356
1567-1364
DOI:10.1111/j.1567-1364.2007.00259.x