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The level of glucose-6-phosphate dehydrogenase activity strongly influences xylose fermentation and inhibitor sensitivity in recombinant Saccharomyces cerevisiae strains

机译:葡萄糖6-磷酸脱氢酶活性水平强烈影响重组酿酒酵母菌株中的木糖发酵和抑制剂敏感性

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摘要

Disruption of the ZWF1 gene encoding glucose-6-phosphate dehydrogenase (G6PDH) has been shown to reduce the xylitol yield and the xylose consumption in the xylose-utilizing recombinant Saccharomyces cerevisiae strain TMB3255. In the present investigation we have studied the influence of different production levels of G6PDH on xylose fermentation. We used a synthetic promoter library and the copper-regulated CUP1 promoter to generate G6PDH-activities between 0% and 179% of the wild- type level. G6PDH-activities of 1% and 6% of the wild-type level resulted in 2.8- and 5.1-fold increase in specific xylose consumption, respectively, compared with the ZWF1-disrupted strain. Both strains exhibited decreased xylitol yields (0.13 and0.19 g/g xylose) and enhanced ethanol yields (0.36 and 0.34 g/g xylose) compared with the control strain TMB3001 (0.29 g xylitol/g xylose, 0.31 g ethanol/g xylose). Cytoplasmic transhydrogenase (TH) from Azotobacter vinelandii has previously been shown to transfer NADPH and NAD+ into NADP+ and NADH, and TH- overproduction resulted in lower xylitol yield and enhanced glycerol yield during xylose utilization. Strains with low G6PDH-activity grew slower in a lignocellulose hydrolysate than the strain with wild-type G6PDH-activity, which suggested that the availability of intracellular NADPH correlated with tolerance towards lignocellulose- derived inhibitors. Low G6PDH-activity strains were also more sensitive to H2 O2 than the control strain TMB3001.
机译:在编码木糖的酿酒酵母菌株TMB3255中,破坏编码6-磷酸葡萄糖脱氢酶(G6PDH)的ZWF1基因可降低木糖醇的产量和木糖的消耗。在本研究中,我们研究了不同生产水平的G6PDH对木糖发酵的影响。我们使用了合成的启动子文库和铜调控的CUP1启动子,以产生介于野生型水平0%至179%之间的G6PDH活性。与破坏ZWF1的菌株相比,G6PDH活性为野生型水平的1%和6%,分别导致特定木糖消耗量增加了2.8倍和5.1倍。与对照菌株TMB3001(0.29 g木糖醇/ g木糖,0.31 g乙醇/ g木糖)相比,两种菌株均表现出降低的木糖醇收率(0.13和0.19 g / g木糖)和提高的乙醇收率(0.36和0.34 g / g木糖)。 。以前已经证明来自葡萄固氮菌的细胞质转氢酶(TH)可以将NADPH和NAD +转移到NADP +和NADH中,TH-的过度生产导致木糖利用过程中木糖醇的收率降低,甘油收率提高。具有低G6PDH活性的菌株在木质纤维素水解物中的生长要比具有野生型G6PDH活性的菌株慢,这表明细胞内NADPH的可用性与对木质纤维素衍生的抑制剂的耐受性相关。低G6PDH活性菌株也比对照菌株TMB3001对H2 O2更为敏感。

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