Cheng Cheng, Tang Rui-Qi, Xiong Liang, Hector Ronald E, Bai Feng-Wu, Zhao Xin-Qing
2School of Life Science and Biotechnology, Dalian University of Technology, Dalian, 116024 China.
1State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240 China.
Biotechnol Biofuels. 2018 Feb 5;11:28. doi: 10.1186/s13068-018-1018-y. eCollection 2018.
wild strains generally have poor xylose-utilization capability, which is a major barrier for efficient bioconversion of lignocellulosic biomass. Laboratory adaption is commonly used to enhance xylose utilization of recombinant . Apparently, yeast cells could remodel the metabolic network for xylose metabolism. However, it still remains unclear why natural isolates of poorly utilize xylose. Here, we analyzed a unique natural isolate YB-2625 which has superior xylose metabolism capability in the presence of mixed-sugar. Comparative transcriptomic analysis was performed using YB-2625 grown in a mixture of glucose and xylose, and the model yeast strain S288C served as a control. Global gene transcription was compared at both the early mixed-sugar utilization stage and the latter xylose-utilization stage.
Genes involved in endogenous xylose-assimilation ( and ), gluconeogenesis, and TCA cycle showed higher transcription levels in YB-2625 at the xylose-utilization stage, when compared to the reference strain. On the other hand, transcription factor encoding genes involved in regulation of glucose repression (, , and ) as well as displayed decreased transcriptional levels in YB-2625, suggesting the alleviation of glucose repression of YB-2625. Notably, genes encoding antioxidant enzymes (, , and ) showed higher transcription levels in YB-2625 in the xylose-utilization stage than that of the reference strain. Consistently, catalase activity of YB-2625 was 1.9-fold higher than that of S288C during the xylose-utilization stage. As a result, intracellular reactive oxygen species levels of YB-2625 were 43.3 and 58.6% lower than that of S288C at both sugar utilization stages. Overexpression of and in the recombinant strain YRH396 deriving from YB-2625 increased cell growth when xylose was used as the sole carbon source, leading to 13.5 and 18.1%, respectively, more xylose consumption.
Enhanced oxidative stress tolerance and relief of glucose repression are proposed to be two major mechanisms for superior xylose utilization by YB-2625. The present study provides insights into the innate regulatory mechanisms underlying xylose utilization in wild-type , which benefits the rapid development of robust yeast strains for lignocellulosic biorefineries.
野生菌株通常木糖利用能力较差,这是木质纤维素生物质高效生物转化的主要障碍。实验室适应性进化常用于提高重组菌的木糖利用能力。显然,酵母细胞可以重塑木糖代谢的代谢网络。然而,目前仍不清楚为什么天然分离株木糖利用能力较差。在此,我们分析了一种独特的天然分离株YB - 2625,其在混合糖存在下具有卓越的木糖代谢能力。使用在葡萄糖和木糖混合物中生长的YB - 2625进行比较转录组分析,并以模式酵母菌株S288C作为对照。在混合糖利用早期阶段和后期木糖利用阶段对全局基因转录进行了比较。
与参考菌株相比,参与内源性木糖同化(和)、糖异生和三羧酸循环的基因在木糖利用阶段的YB - 2625中显示出更高的转录水平。另一方面,表示参与葡萄糖阻遏调控的转录因子编码基因(、和)以及在YB - 2625中的转录水平降低,表明YB - 2625的葡萄糖阻遏得到缓解。值得注意的是,编码抗氧化酶(、、和)的基因在木糖利用阶段的YB - 2625中的转录水平高于参考菌株。一致地,在木糖利用阶段,YB - 2625的过氧化氢酶活性比S288C高1.9倍。结果,在两个糖利用阶段,YB - 2625的细胞内活性氧水平分别比S288C低43.3%和58.6%。在源自YB - 2625的重组菌株YRH396中过表达和,当以木糖作为唯一碳源时增加了细胞生长,分别导致木糖消耗增加13.5%和18.1%。
增强的氧化应激耐受性和葡萄糖阻遏的缓解被认为是YB - 2625卓越木糖利用的两个主要机制。本研究为野生型木糖利用的内在调控机制提供了见解,这有利于木质纤维素生物炼制中强大酵母菌株的快速开发。