在木糖培养基中,删除JEN1和ADY2基因可降低表达异源乳酸脱氢酶的工程酿酒酵母的乳酸产量。

Deletion of JEN1 and ADY2 reduces lactic acid yield from an engineered Saccharomyces cerevisiae, in xylose medium, expressing a heterologous lactate dehydrogenase.

作者信息

Turner Timothy L, Lane Stephan, Jayakody Lahiru N, Zhang Guo-Chang, Kim Heejin, Cho Whiyeon, Jin Yong-Su

机构信息

Department of Food Science and Human Nutrition, 260 Bevier Hall, 905 South Goodwin Avenue, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

Carl R. Woese Institute for Genomic Biology, 1206 West Gregory Avenue, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

出版信息

FEMS Yeast Res. 2019 Sep 1;19(6). doi: 10.1093/femsyr/foz050.

Abstract

Microorganisms have evolved to produce specific end products for many reasons, including maintaining redox balance between NAD+ and NADH. The yeast Saccharomyces cerevisiae, for example, produces ethanol as a primary end product from glucose for the regeneration of NAD+. Engineered S. cerevisiae strains have been developed to ferment lignocellulosic sugars, such as xylose, to produce lactic acid by expression of a heterologous lactate dehydrogenase (ldhA from Rhizopus oryzae) without genetic perturbation to the native ethanol pathway. Surprisingly, the engineered yeast strains predominantly produce ethanol from glucose, but produce lactic acid as the major product from xylose. Here, we provide initial evidence that the shift in product formation from ethanol to lactic acid during xylose fermentation is at least partially dependent on the presence of functioning monocarboxylate transporter genes/proteins, including JEN1 and ADY2, which are downregulated and unstable in the presence of glucose, but upregulated/stable on xylose. Future yeast metabolic engineering studies may find the feedstock/carbon selection, such as xylose, an important step toward improving the yield of target end products.

摘要

微生物出于多种原因进化出产生特定终产物的能力,包括维持NAD⁺和NADH之间的氧化还原平衡。例如,酿酒酵母会将乙醇作为葡萄糖的主要终产物来再生NAD⁺。通过表达来自米根霉的异源乳酸脱氢酶(ldhA),已构建出工程化的酿酒酵母菌株来发酵木质纤维素糖(如木糖)以产生乳酸,同时不对天然乙醇代谢途径进行基因干扰。令人惊讶的是,这些工程化酵母菌株在葡萄糖发酵时主要产生乙醇,但在木糖发酵时以乳酸作为主要产物。在此,我们提供了初步证据,表明木糖发酵过程中产物形成从乙醇向乳酸的转变至少部分依赖于功能性单羧酸转运蛋白基因/蛋白质的存在,包括JEN1和ADY2,它们在葡萄糖存在时表达下调且不稳定,但在木糖上表达上调/稳定。未来的酵母代谢工程研究可能会发现原料/碳源选择(如木糖)是提高目标终产物产量的重要一步。

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