2C 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 (m) for E. coli P84A/MC1061 were promoted in the log phase (4-8 h) under appropriate Zn 2C 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 max ) was increased by 9.80%, and the time to reach HheC max was reduced from 16 to 12 hours. Furthermore, appropriate supplementation of Zn 2C caused higher m 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.
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