Enhancing the tolerance of Clostridium saccharoperbutylacetonicum to lignocellulosic-biomass-derived inhibitors for efficient biobutanol production by overexpressing efflux pumps genes from Pseudomonas putida
Enhancing the tolerance of Clostridium saccharoperbutylacetonicum to lignocellulosic-biomass-derived inhibitors for efficient biobutanol production by overexpressing efflux pumps genes from Pseudomonas putida
复制标题
DOI：
10.1016/j.biortech.2020.123532
复制
发表时间：
2020-09-01
影响因子：
11.4
通讯作者：
Wang, Yi
中科院分区：
文献类型：
--
作者：
Jimenez-Bonilla, Pablo;Zhang, Jie;Wang, Yi
生物质预处理过程中产生的呋喃醛和酚类化合物会抑制发酵生产生物燃料。外排泵主动将小分子输送出细胞，从而维持正常的微生物新陈代谢。恶臭假单胞菌对丁醇和其他小分子具有突出的耐受性，我们推测它的外排泵可能对这种稳健性起着至关重要的作用。在这里，我们从恶臭假单胞菌中过表达外排泵基因，以提高生产高丁醇的丙酮酸梭菌对发酵抑制剂的耐受性。有趣的是，整个单位的过度表达导致耐受性降低，而单是亚单位(SrpB)的过度表达显著增强了菌株的稳健性。与对照相比，工程菌在含有17%多呋喃或50%阿魏酸的培养基中生长能力增强，产量接近14g/L丁醇(与常规条件下不加抑制剂的发酵相当)。本研究为提高微生物对木质纤维原料高效生产生物燃料的稳健性提供了有价值的参考。
Furan aldehydes and phenolic compounds generated during biomass pretreatment can inhibit fermentation for biofuel production. Efflux pumps actively transport small molecules out of cells, thus sustaining normal microbial metabolism. Pseudomonas putida has outstanding tolerance to butanol and other small molecules, and we hypothesize that its efflux pump could play essential roles for such robustness. Here, we overexpressed efflux pump genes from P. putida to enhance tolerance of hyper-butanol producing Clostridium saccharoperbutylacetonicum to fermentation inhibitors. Interestingly, overexpression of the whole unit resulted in decreased tolerance, while overexpression of the subunit (srpB) alone exerted significant enhanced robustness of the strain. Compared to the control, the engineered strain had enhanced capability to grow in media containing 17% more furfural or 50% more ferulic acid, and produced similar to 14 g/L butanol (comparable to fermentation under regular conditions without inhibitors). This study provided valuable reference for boosting microbial robustness towards efficient biofuel production from lignocellulosic materials.