School of Life Science and Biotechnology, Dalian University of Technology, Dalian, 116024, China.
State Key Laboratory of Microbial Metabolism, Shanghai Jiaotong University, Shanghai, 200240, China.
Microb Cell Fact. 2017 Oct 30;16(1):181. doi: 10.1186/s12934-017-0795-5.
Multiple lignocellulose-derived inhibitors represent great challenges for bioethanol production from lignocellulosic materials. These inhibitors that are related to the levels of intracellular reactive oxidative species (ROS) make oxidoreductases a potential target for an enhanced tolerance in yeasts.
In this study, the thioredoxin and its reductase from Kluyveromyces marxianus Y179 was identified, which was subsequently achieved over-expression in Saccharomyces cerevisiae 280. In spite of the negative effects by expression of thioredoxin gene (KmTRX), the thioredoxin reductase (KmTrxR) helped to enhance tolerance to multiple lignocellulose-derived inhibitors, such as formic acid and acetic acid. In particular, compared with each gene expression, the double over-expression of KmTRX2 and KmTrxR achieved a better ethanol fermentative profiles under a mixture of formic acid, acetic acid, and furfural (FAF) with a shorter lag period. At last, the mechanism that improves the tolerance depended on a normal level of intracellular ROS for cell survival under stress.
The synergistic effect of KmTrxR and KmTRX2 provided the potential possibility for ethanol production from lignocellulosic materials, and give a general insight into the possible toxicity mechanisms for further theoretical research.
多种木质纤维素衍生抑制剂对木质纤维素材料的生物乙醇生产构成了巨大挑战。这些与细胞内活性氧化物质 (ROS) 水平相关的抑制剂使氧化还原酶成为提高酵母耐受性的潜在目标。
本研究鉴定了马克斯克鲁维酵母 Y179 的硫氧还蛋白及其还原酶,并在酿酒酵母 280 中实现了过表达。尽管硫氧还蛋白基因 (KmTRX) 的表达产生了负面影响,但硫氧还蛋白还原酶 (KmTrxR) 有助于提高对多种木质纤维素衍生抑制剂(如甲酸和乙酸)的耐受性。特别是与每个基因表达相比,KmTRX2 和 KmTrxR 的双重过表达在甲酸、乙酸和糠醛 (FAF) 的混合物下实现了更好的乙醇发酵谱,具有更短的迟滞期。最后,提高耐受性的机制依赖于细胞在应激下正常的细胞内 ROS 水平以实现细胞存活。
KmTrxR 和 KmTRX2 的协同作用为木质纤维素材料的乙醇生产提供了潜在的可能性,并为进一步的理论研究提供了对可能毒性机制的全面了解。