A baker's yeast mutant (fil1) with a specific, partially inactivating mutation in adenylate cyclase maintains a high stress resistance during active fermentation and growth.
The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in stress resistance. This is disadvantageous for several biotechnological applications, e.g. the preparation of freeze doughs. We have isolated mutants in a laboratory strain which are deficient in fermentation-induced loss of stress resistance ('fil' mutants) using a heat shock selection protocol. We show that the fil1 mutant contains a mutation in the CYR1 gene which encodes adenylate cyclase. It causes a change at position 1682 of glutamate into lysine and results in a tenfold drop in adenylate cyclase activity. The fil1 mutant displays a reduction in the glucose-induced cAMP increase, trehalase activation and loss of heat resistance. Interestingly, the fil1 mutant shows the same growth and fermentation rate as the wild type strain, as opposed to other mutants with reduced activity of the cAMP pathway. Introduction of the fil1 mutation in the vigorous Y55 strain and cultivation of the mutant under pilot scale conditions resulted in a yeast that displayed a higher freeze and drought resistance during active fermentation compared to the wild type Y55 strain. These results show that high stress resistance and high fermentation activity are compatible biological properties. Isolation of fil-type mutations appears a promising avenue for development of industrial yeast strains with improved stress resistance during active fermentation.
Van Dijck
P
P
Laboratorium voor Moleculaire Celbiologie, Institute of Botany and Microbiology, Katholieke Universiteit Leuven, Flanders, Belgium.
Ma
P
P
Versele
M
M
Gorwa
M F
MF
Colombo
S
S
Lemaire
K
K
Bossi
D
D
Loïez
A
A
Thevelein
J M
JM
D000262:Adenylyl Cyclases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000242:Cyclic AMP / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D005020:Ethyl Methanesulfonate; D005285:Fermentation; D005808:Genes, Recessive; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid; D007700:Kinetics; D008239:Lysine; D016296:Mutagenesis; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D016415:Sequence Alignment; D013696:Temperature
D000431:Ethanol; D018698:Glutamic Acid; D005020:Ethyl Methanesulfonate; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D005947:Glucose; D008239:Lysine