State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.
Biotechnol Prog. 2010 Nov-Dec;26(6):1551-7. doi: 10.1002/btpr.496. Epub 2010 Sep 30.
This study aimed to increase the glycolytic flux of the multivitamin auxotrophic yeast Torulopsis glabrata by redirecting NADH oxidation from oxidative phosphorylation to membrane-bound ferric reductase. We added potassium ferricyanide as electron acceptor to T. glabrata culture broth at 20% dissolved oxygen (DO) concentration, which resulted in: (1) decreases in the NADH content, NADH/NAD(+) ratio, and ATP level of 45.3%, 60.3%, and 15.2%, respectively; (2) high activities of the key glycolytic enzymes hexokinase, phosphofructokinase, and pyruvate kinase, as well as high expression levels of the genes encoding these enzymes; and (3) increases in the specific glucose consumption rate and pyruvate yield of T. glabrata was by 45.5% and 23.1%, respectively. Our results showed that membrane-bound ferric reductase offers an alternative and efficient NADH oxidation pathway at lower DO concentration, which increases the glycolytic flux of T. glabrata.
本研究旨在通过将 NADH 氧化从氧化磷酸化重定向到膜结合的铁还原酶来增加多维生素营养缺陷型酵母光滑球拟酵母的糖酵解通量。我们在 20%溶解氧(DO)浓度下向光滑球拟酵母培养液中添加铁氰化钾作为电子受体,结果导致:(1)NADH 含量、NADH/NAD+ 比和 ATP 水平分别降低了 45.3%、60.3%和 15.2%;(2)关键糖酵解酶己糖激酶、磷酸果糖激酶和丙酮酸激酶的高活性以及编码这些酶的基因的高表达水平;以及(3)光滑球拟酵母的比葡萄糖消耗速率和丙酮酸产率分别提高了 45.5%和 23.1%。我们的结果表明,在较低的 DO 浓度下,膜结合的铁还原酶提供了一种替代的、有效的 NADH 氧化途径,从而增加了光滑球拟酵母的糖酵解通量。