Cholet Orianne, Hénaut Alain, Hébert Agnès, Bonnarme Pascal
UMR782 Génie et Microbiologie des Procédés Alimentaires, AgroParisTech, INRA, F-78850 Thiverval Grignon, France.
Appl Environ Microbiol. 2008 Jun;74(11):3356-67. doi: 10.1128/AEM.00644-07. Epub 2008 Apr 4.
Yarrowia lipolytica is one of the yeasts most frequently isolated from the surface of ripened cheeses. In previous work, it has been shown that this yeast is able to convert L-methionine into various volatile sulfur compounds (VSCs) that may contribute to the typical flavors of several cheeses. In the present study, we show that Y. lipolytica does not assimilate lactate in the presence of L-methionine in a cheeselike medium. Nineteen presumptive genes associated with L-methionine catabolism or pyruvate metabolism in Y. lipolytica were transcriptionally studied in relation to L-methionine degradation. The expression levels of the YlARO8 (YALI0E20977g), YlBAT1 (YALI0D01265g), and YlBAT2 (YALI0F19910g) genes (confirmed by real-time PCR experiments) were found to be strongly up-regulated by L-methionine, and a greater variety and larger amounts of VSCs, such as methanethiol and its autooxidation products (dimethyl disulfide and dimethyl trisulfide), were released in the medium when Y. lipolytica was grown in the presence of a high concentration of L-methionine. In contrast, other genes related to pyruvate metabolism were found to be down-regulated in the presence of L-methionine; two exceptions were the YlPDB1 (YALI0E27005g) and YlPDC6 (YALI0D06930g) genes, which encode a pyruvate dehydrogenase and a pyruvate decarboxylase, respectively. Both transcriptional and biochemical results corroborate the view that transamination is the first step of the enzymatic conversion of L-methionine to VSCs in Y. lipolytica and that the YlARO8, YlBAT1, and YlBAT2 genes could play a key role in this process.
解脂耶氏酵母是最常从成熟奶酪表面分离出的酵母之一。在先前的研究中,已表明这种酵母能够将L-甲硫氨酸转化为多种挥发性硫化合物(VSC),这些化合物可能有助于几种奶酪的典型风味。在本研究中,我们发现解脂耶氏酵母在类似奶酪的培养基中,在L-甲硫氨酸存在的情况下不会同化乳酸。针对与解脂耶氏酵母中L-甲硫氨酸分解代谢或丙酮酸代谢相关的19个推定基因,研究了其与L-甲硫氨酸降解相关的转录情况。发现YlARO8(YALI0E20977g)、YlBAT1(YALI0D01265g)和YlBAT2(YALI0F19910g)基因的表达水平(通过实时PCR实验证实)在L-甲硫氨酸作用下强烈上调,并且当解脂耶氏酵母在高浓度L-甲硫氨酸存在下生长时,培养基中会释放出更多种类和更大数量的VSC,如甲硫醇及其自氧化产物(二甲基二硫和二甲基三硫)。相反,发现与丙酮酸代谢相关的其他基因在L-甲硫氨酸存在下被下调;两个例外是YlPDB1(YALI0E27005g)和YlPDC6(YALI0D06930g)基因,它们分别编码丙酮酸脱氢酶和丙酮酸脱羧酶。转录和生化结果均证实了以下观点:转氨作用是解脂耶氏酵母中L-甲硫氨酸酶促转化为VSC的第一步,并且YlARO8、YlBAT1和YlBAT2基因可能在这一过程中起关键作用。