Zhang Qin, You Shaolin, Li Yanbin, Qu Xiaowei, Jiang Hui
College of Biological and Chemical Engineering, Anhui Polytechnic University, Wuhu, 241000 Anhui China.
College of Life Science, Tarim University, Alaer, 843300 Xinjiang China.
Biotechnol Biofuels. 2020 May 22;13:94. doi: 10.1186/s13068-020-01733-9. eCollection 2020.
Biohydrogen production from lignocellulose has become an important hydrogen production method due to its diversity, renewability, and cheapness. Overexpression of the formate hydrogen lyase activator () gene is a promising tactic for enhancement of hydrogen production in facultative anaerobic . As a species of , was reported as a highly efficient hydrogen-producing bacterium. However, little work has been reported in terms of cloning and expressing the gene in for lignocellulose-based hydrogen production.
In this study, the formate hydrogen lyase activator () gene was cloned and overexpressed in WL1318. We found that the recombinant strain significantly enhanced cumulative hydrogen production by 188% following fermentation of cotton stalk hydrolysate for 24 h, and maintained improved production above 30% throughout the fermentation process compared to the wild strain. Accordingly, overexpression of the gene resulted in an enhanced hydrogen production potential () and maximum hydrogen production rate ( ), as well as a shortened lag phase time () for the recombinant strain. Additionally, the recombinant strain also displayed improved glucose (12%) and xylose (3.4%) consumption and hydrogen yield Y(H/S) (37.0%) compared to the wild strain. Moreover, the metabolites and specific enzyme profiles demonstrated that reduced flux in the competitive branch, including succinic, acetic, and lactic acids, and ethanol generation, coupled with increased flux in the pyruvate node and formate splitting branch, benefited hydrogen synthesis.
The results conclusively prove that overexpression of gene in WL1318 can effectively enhance the hydrogen production from cotton stalk hydrolysate, and reduce the metabolic flux in the competitive branch. It is the first attempt to engineer the gene in the hydrogen-producing bacterium . This work provides a highly efficient engineered bacterium for biohydrogen production from fermentation of lignocellulosic hydrolysate in the future.
由于木质纤维素具有多样性、可再生性和廉价性,利用其生产生物氢已成为一种重要的制氢方法。甲酸氢裂解酶激活剂()基因的过表达是提高兼性厌氧菌产氢能力的一种有前景的策略。作为的一个物种,被报道为一种高效产氢细菌。然而,关于在中克隆和表达用于基于木质纤维素的产氢的基因的研究报道较少。
在本研究中,甲酸氢裂解酶激活剂()基因在WL1318中被克隆并过表达。我们发现,重组菌株在棉秆水解物发酵24小时后,累积产氢量显著提高了188%,并且在整个发酵过程中与野生菌株相比,产氢量保持提高30%以上。因此,基因的过表达导致重组菌株的产氢潜力()和最大产氢速率()提高,以及延迟期时间()缩短。此外,与野生菌株相比,重组菌株的葡萄糖(12%)和木糖(3.4%)消耗量以及氢产率Y(H/S)(37.0%)也有所提高。此外,代谢物和特定酶谱表明,竞争分支(包括琥珀酸、乙酸和乳酸以及乙醇生成)中的通量降低,同时丙酮酸节点和甲酸裂解分支中的通量增加,有利于氢的合成。
结果确凿地证明,在WL1318中过表达基因可以有效地提高棉秆水解物产氢能力,并降低竞争分支中的代谢通量。这是首次尝试在产氢细菌中对基因进行工程改造。这项工作为未来从木质纤维素水解物发酵生产生物氢提供了一种高效的工程菌。