Mansour S, Beckerich J M, Bonnarme P
Agro Paris Tech-INRA, UMR 782 Génie et Microbiologie des Procédés Alimentaires, 78850 Thiverval Grignon, France.
Appl Environ Microbiol. 2008 Nov;74(21):6505-12. doi: 10.1128/AEM.01519-08. Epub 2008 Sep 5.
The consumption of lactate and amino acids is very important for microbial development and/or aroma production during cheese ripening. A strain of Yarrowia lipolytica isolated from cheese was grown in a liquid medium containing lactate in the presence of a low (0.1x) or high (2x) concentration of amino acids. Our results show that there was a dramatic increase in the growth of Y. lipolytica in the medium containing a high amino acid concentration, but there was limited lactate consumption. Conversely, lactate was efficiently consumed in the medium containing a low concentration of amino acids after amino acid depletion was complete. These data suggest that the amino acids are used by Y. lipolytica as a main energy source, whereas lactate is consumed following amino acid depletion. Amino acid degradation was accompanied by ammonia production corresponding to a dramatic increase in the pH. The effect of adding amino acids to a Y. lipolytica culture grown on lactate was also investigated. Real-time quantitative PCR analyses were performed with specific primers for five genes involved in amino acid transport and catabolism, including an amino acid transporter gene (GAP1) and four aminotransferase genes (ARO8, ARO9, BAT1, and BAT2). The expression of three genes involved in lactate transport and catabolism was also studied. These genes included a lactate transporter gene (JEN1) and two lactate dehydrogenase genes (CYB2-1 and CYB2-2). Our data showed that GAP1, BAT2, BAT1, and ARO8 were maximally expressed after 15 to 30 min following addition of amino acids (BAT2 was the most highly expressed gene), while the maximum expression of JEN1, CYB2-1, and CYB2-2 was delayed (>or=60 min).
在奶酪成熟过程中,乳酸和氨基酸的消耗对于微生物生长及/或香气产生非常重要。从奶酪中分离出的一株解脂耶氏酵母在含有乳酸的液体培养基中培养,培养基中氨基酸浓度分别为低浓度(0.1x)或高浓度(2x)。我们的结果表明,在氨基酸浓度高的培养基中,解脂耶氏酵母的生长显著增加,但乳酸消耗有限。相反,在氨基酸耗尽后,低浓度氨基酸培养基中的乳酸被有效消耗。这些数据表明,解脂耶氏酵母将氨基酸作为主要能量来源,而在氨基酸耗尽后才消耗乳酸。氨基酸降解伴随着氨的产生,这导致pH值急剧升高。我们还研究了向以乳酸为生长底物的解脂耶氏酵母培养物中添加氨基酸的影响。使用参与氨基酸转运和分解代谢的五个基因的特异性引物进行实时定量PCR分析,这五个基因包括一个氨基酸转运蛋白基因(GAP1)和四个转氨酶基因(ARO8、ARO9、BAT1和BAT2)。我们还研究了参与乳酸转运和分解代谢的三个基因的表达。这些基因包括一个乳酸转运蛋白基因(JEN1)和两个乳酸脱氢酶基因(CYB2-1和CYB2-2)。我们的数据表明,添加氨基酸后15至30分钟,GAP1、BAT2、BAT1和ARO8的表达达到最大值(BAT2是表达量最高的基因),而JEN1、CYB2-1和CYB2-2的最大表达延迟(≥60分钟)。