Minard Karyl I, McAlister-Henn L
Department of Biochemistry, University of Texas Health Science Center, San Antonio, 78229-3900, USA.
Arch Biochem Biophys. 2009 Mar 1;483(1):136-43. doi: 10.1016/j.abb.2008.12.014. Epub 2008 Dec 30.
Following a shift to medium with acetate as the carbon source, a parental yeast strain exhibited a transient moderate 20% reduction in total cellular [NAD(+)+NADH] but showed a approximately 10-fold increase in the ratio of [NAD(+)]:[NADH] after 36h. A mutant strain (idhDelta) lacking the tricarboxylic acid cycle enzyme isocitrate dehydrogenase had 50% higher cellular levels of [NAD(+)+NADH] relative to the parental strain but exhibited similar changes in cofactor concentrations following a shift to acetate medium, despite an inability to grow on that carbon source; essentially all of the cofactor was in the oxidized form within 36h. The salvage pathway for NAD(H) biosynthesis was found to be particularly important for viability during early transition of the parental strain to stationary phase in acetate medium. However, oxygen consumption was not affected, suggesting that the NAD(H) produced during this time may support other cellular functions. The idhDelta mutant exhibited increased flux through the salvage pathway in acetate medium but was dependent on the de novo pathway for viability. Long-term chronological lifespans of the parental and idhDelta strains were similar, but viability of the mutant strain was dependent on both pathways for NAD(H) biosynthesis.
在转换为以乙酸盐作为碳源的培养基后,亲本酵母菌株的细胞总[NAD(+)+NADH]含量出现短暂适度下降,降低了20%,但在36小时后,[NAD(+)]:[NADH]的比值增加了约10倍。缺乏三羧酸循环酶异柠檬酸脱氢酶的突变菌株(idhDelta),其细胞内[NAD(+)+NADH]水平相对于亲本菌株高50%,但在转换到乙酸盐培养基后,尽管无法在该碳源上生长,其辅因子浓度仍表现出类似变化;基本上所有辅因子在36小时内都处于氧化形式。发现NAD(H)生物合成的补救途径对于亲本菌株在乙酸盐培养基中早期过渡到稳定期的生存能力尤为重要。然而,氧气消耗未受影响,这表明在此期间产生的NAD(H)可能支持其他细胞功能。idhDelta突变体在乙酸盐培养基中通过补救途径的通量增加,但生存能力依赖于从头合成途径。亲本菌株和idhDelta菌株的长期衰老寿命相似,但突变菌株的生存能力依赖于NAD(H)生物合成的两条途径。