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Appl Environ Microbiol. 1991 October; 57(10): 2796-2803
Copyright © 1991, American Society for Microbiology. All Rights Reserved.

Chromosomal Integration and Expression of Two Bacterial -Acetolactate Decarboxylase Genes in Brewer's Yeast

K. Blomqvist, M.-L. Suihko^*, J. Knowles and M. Penttilä

Biotechnical Laboratory, Technical Research Centre of Finland (VTT), P.O. Box 202, SF-02151 Espoo, Finland

ABSTRACT

A bacterial gene encoding -acetolactate decarboxylase, isolated from Klebsiella terrigena or Enterobacter aerogenes, was expressed in brewer's yeast. The genes were expressed under either the yeast phosphoglycerokinase (PGK1) or the alcohol dehydrogenase (ADH1) promoter and were integrated by gene replacement by using cotransformation into the PGK1 or ADH1 locus, respectively, of a brewer's yeast. The expression level of the -acetolactate decarboxylase gene of the PGK1 integrant strains was higher than that of the ADH1 integrants. Under pilot-scale brewing conditions, the -acetolactate decarboxylase activity of the PGK1 integrant strains was sufficient to reduce the formation of diacetyl below the taste threshold value, and no lagering was needed. The brewing properties of the recombinant yeast strains were otherwise unaltered, and the quality (most importantly, the flavor) of the trial beers produced was as good as that of the control beer.

^* Corresponding author.

Present address: Institute of Microbiology, University of Umeå, Umeå, Sweden.

Present address: Glaxo Institute for Molecular Biology, Geneva, Switzerland.

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