3 Biotech ( IF 2.8 ) Pub Date : 2021-08-10 , DOI: 10.1007/s13205-021-02959-6 Peng Yang 1 , Yanan Chen 1 , An-Dong Gong 1
Corynebacterium glutamicum has been widely used for bulk and fine chemicals fermentation these years. In this study, we developed a defined medium for this bacteria based on the widely used CGXII minimal medium. We evaluated the effects of different components in CGXII on cell growth of C. glutamicum ATCC 13032 and improved the medium through single-factor experiment and central composite design (CCD). Urea, K2HPO4 and MgSO4 were found to be significant factors. 7 out of the total 15 components were modified. (NH4)2SO4, KH2PO4, and protocatechuic acid were eliminated. Amounts of urea and MgSO4 were increased, and concentrations of biotin and glucose were reduced. The resulting R2 medium was proved to be more suitable for cell growth, plasmid amplification and protein production than the original recipe. Remarkably, cell biomass accumulation in R2 increased by 54.36% than CGXII. Transcriptome analysis revealed alteration of carbon metabolism, cation transport and energy synthesis, which might be beneficial for cell growth in R2. Considering the high nitrogen content and availability of urea, the new medium is simplified and cost effective, which holds attractive potential for future study.
中文翻译:
以尿素为氮源的谷氨酸棒杆菌确定培养基的开发
近年来,谷氨酸棒杆菌被广泛用于大宗和精细化学品的发酵。在这项研究中,我们基于广泛使用的 CGXII 基本培养基开发了一种用于这种细菌的明确培养基。我们评估了 CGXII 中不同成分对谷氨酸棒杆菌ATCC 13032 细胞生长的影响,并通过单因素实验和中心复合设计 (CCD) 改进了培养基。尿素、K 2 HPO 4和MgSO 4被发现是重要因素。总共 15 个组件中的 7 个进行了修改。(NH 4 ) 2 SO 4、KH 2 PO 4和原儿茶酸被消除。尿素和 MgSO 的量4增加,生物素和葡萄糖的浓度降低。由此产生的 R2 培养基被证明比原始配方更适合细胞生长、质粒扩增和蛋白质生产。值得注意的是,R2 中的细胞生物量积累比 CGXII 增加了 54.36%。转录组分析揭示了碳代谢、阳离子转运和能量合成的改变,这可能有利于 R2 中的细胞生长。考虑到尿素的高氮含量和可用性,新培养基简化且具有成本效益,具有未来研究的诱人潜力。