Van Dijck P, Ma P, Versele M, Gorwa M F, Colombo S, Lemaire K, Bossi D, Loïez A, Thevelein J M
Laboratorium voor Moleculaire Celbiologie, Institute of Botany and Microbiology, Katholieke Universiteit Leuven, Flanders, Belgium.
J Mol Microbiol Biotechnol. 2000 Oct;2(4):521-30.
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.
酿酒酵母中发酵的启动与抗逆性的迅速下降有关。这对几种生物技术应用是不利的,例如冷冻面团的制备。我们使用热休克选择方案在实验室菌株中分离出了在发酵诱导的抗逆性丧失方面存在缺陷的突变体(“fil”突变体)。我们表明,fil1突变体在编码腺苷酸环化酶的CYR1基因中存在一个突变。它导致第1682位的谷氨酸变为赖氨酸,并使腺苷酸环化酶活性下降十倍。fil1突变体表现出葡萄糖诱导的cAMP增加、海藻糖酶激活和耐热性丧失减少。有趣的是,与cAMP途径活性降低的其他突变体相反,fil1突变体的生长和发酵速率与野生型菌株相同。将fil1突变引入活力旺盛的Y55菌株并在中试规模条件下培养该突变体,得到的酵母在活跃发酵期间表现出比野生型Y55菌株更高的抗冻和抗旱性。这些结果表明,高抗逆性和高发酵活性是兼容的生物学特性。分离fil型突变似乎是开发在活跃发酵期间具有改善抗逆性的工业酵母菌株的一条有前途的途径。