Li Xiao, Luo Yu-Di, Pan Dong-Rui, Shi Xiao-Dan, Tan Ya-Li, Li Zhi-Hong
a College of Biological and Pharmaceutica Engineering, China Three Gorges University , Yichang , P. R. China.
b Angel Yeast Co., Ltd. Yichang , P. R. China.
Bioengineered. 2017 Sep 3;8(5):585-593. doi: 10.1080/21655979.2017.1285985. Epub 2017 Mar 10.
Using 5 Zn supplementation strategies in a 50 L batch bioreactor named FUS-50L(A), possible correlations among Zn content and addition timing, physiologic activity (PA), halohydrin dehalogenase (HheC) accumulation of Escherichia coli P84A/MC1061 were systematically investigated. First, Zn was confirmed as the significant factor, and its optimal concentration for HheC expression was 3.87 mg/L through fermentation experiments in shaking flasks. Second, based on experimental results from the different strategies, it was found that PA, nutrient consumption rate (NCR) and specific growth rate (μ) for E. coli P84A/MC1061 were promoted in the log phase (4-8 h) under appropriate Zn concentrations in the lag phase and late log phase. Furthermore cell biomass was also increased to a higher level and the maximum HheC activity (i.e. HheC) was increased by 9.80%, and the time to reach HheC was reduced from 16 to 12 hours. Furthermore, appropriate supplementation of Zn caused higher μ for E. coli P84A/MC1061, which resulted in more rapid accumulation of increased acetic acid concentrations, leading to higher acetic acid consumption avoiding any negative effects on producing HheC because of carbon source being exhausted prematurely and acetic acid being consumed rapidly.
在名为FUS-50L(A)的50升分批生物反应器中采用5种锌补充策略,系统研究了锌含量与添加时间、生理活性(PA)、大肠杆菌P84A/MC1061的卤代醇脱卤酶(HheC)积累之间的可能相关性。首先,通过摇瓶发酵实验确定锌是显著因素,其对HheC表达的最佳浓度为3.87毫克/升。其次,基于不同策略的实验结果发现,在延迟期和对数后期适当的锌浓度下,大肠杆菌P84A/MC1061的PA、营养物质消耗率(NCR)和比生长速率(μ)在对数期(4-8小时)得到促进。此外,细胞生物量也增加到更高水平,最大HheC活性(即HheC)提高了9.80%,达到HheC的时间从16小时减少到12小时。此外,适当补充锌使大肠杆菌P84A/MC1061的μ更高,这导致乙酸浓度增加的积累更快,从而避免了由于碳源过早耗尽和乙酸快速消耗而对产生HheC产生任何负面影响,同时实现了更高的乙酸消耗。