Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China.
SynBio Research Platform, Collaborative Innovation Centre of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China.
Sci Rep. 2017 Apr 25;7:46759. doi: 10.1038/srep46759.
Adaptive evolution by serial subcultivation of co-cultured Bacillus thuringiensis and Ketogulonicigenium vulgare significantly enhanced the productivity of 2-keto-L-gulonic acid in two-step vitamin C production. The adaptation mechanism in K. vulgare-B. thuringiensis consortium was investigated in this study based on comparative genomics and metabolomics studies. It was found that the growth, anti-oxidation, transcription and regulation were significantly enhanced in the adapted consortium. The mutation of the genes, which encode amidohydrolase in adapted K. vulgare (K150) and amino acid permease in adapted B. thuringiensis (B150), resulted in the increase of some amino acids levels in each species, and further enhanced the metabolic exchange and growth ability of the two species. Besides, the mutation of the gene encoding spore germination protein enhanced the metabolic levels of tricarboxylic acid cycle, and decreased the sporulation in B150, which induced its growth. The mutation of the genes, which encode NADPH nitroreductase in K150 and NADPH-dependent FMN reductase in B150, may enhance the ability of anti-oxidation. Overall, the long-term adaptation of K. vulgare and B. thuringiensis influenced the global regulation and made them more inseparable in metabolite exchange. Our work will provide ideas for the molecular design and optimization in microbial consortium.
通过共培养的苏云金芽孢杆菌和酮古龙酸菌的连续亚培养进行适应性进化,显著提高了两步法维生素 C 生产中 2-酮-L-古洛糖酸的产量。本研究基于比较基因组学和代谢组学研究,探讨了酮古龙酸菌-苏云金芽孢杆菌联合体的适应机制。结果发现,适应联合体中的生长、抗氧化、转录和调控都得到了显著增强。适应的酮古龙酸菌(K150)和适应的苏云金芽孢杆菌(B150)中编码酰胺水解酶和氨基酸渗透酶的基因发生突变,导致每个物种中的一些氨基酸水平增加,进一步增强了两种物种的代谢交换和生长能力。此外,编码孢子萌发蛋白的基因发生突变,增强了三羧酸循环的代谢水平,减少了 B150 中的孢子形成,从而促进了其生长。K150 中编码 NADPH 硝基还原酶和 B150 中编码 NADPH 依赖性 FMN 还原酶的基因发生突变,可能增强了抗氧化能力。总的来说,酮古龙酸菌和苏云金芽孢杆菌的长期适应影响了全局调控,使它们在代谢物交换中更加不可分割。我们的工作将为微生物联合体的分子设计和优化提供思路。