Nucleotide sequence and expression of the Enterobacter aerogenes alpha-acetolactate decarboxylase gene in brewer's yeast.
The nucleotide sequence of a 1.4-kilobase DNA fragment containing the alpha-acetolactate decarboxylase gene of Enterobacter aerogenes was determined. The sequence contains an entire protein-coding region of 780 nucleotides which encodes an alpha-acetolactate decarboxylase of 260 amino acids. The DNA sequence coding for alpha-acetolactate decarboxylase was placed under the control of the alcohol dehydrogenase I promoter of the yeast Saccharomyces cerevisiae in a plasmid capable of autonomous replication in both S. cerevisiae and Escherichia coli. Brewer's yeast cells transformed by this plasmid showed alpha-acetolactate decarboxylase activity and were used in laboratory-scale fermentation experiments. These experiments revealed that the diacetyl concentration in wort fermented by the plasmid-containing yeast strain was significantly lower than that in wort fermented by the parental strain. These results indicated that the alpha-acetolactate decarboxylase activity produced by brewer's yeast cells degraded alpha-acetolactate and that this degradation caused a decrease in diacetyl production.
Sone
H
H
Central Laboratories of Key Technology, Kirin Brewery Co., Ltd., Gunma, Japan.
Fujii
T
T
Kondo
K
K
Shimizu
F
F
Tanaka
J
J
Inoue
T
T
D001483:Base Sequence; D002262:Carboxy-Lyases / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D004754:Enterobacter / Q000201:enzymology* / Q000235:genetics; D004755:Enterobacteriaceae / Q000201:enzymology*; D005285:Fermentation; D005798:Genes, Bacterial; D007773:Lactates / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D007773:Lactates; C006359:alpha-acetolactate; D002262:Carboxy-Lyases; C023746:acetolactate decarboxylase