Abstract
Microbial fermentation for L-methionine (L-Met) production based on natural renewable resources is attractive and challenging. In this work, the effects of medium composition and fermentation conditions were investigated to improve L-Met production by genetically engineered Escherichia coli MET-3. Statistical optimization techniques including Plackett–Burman (PB) design and Box–Behnken design (BBD) were adopted first to optimize the culture medium. Results of PB-designed experiments indicated that the culture medium components including glucose, yeast extract, KH2PO4, and MgSO4.7H2O had significant effects on L-Met biosynthesis. With their best-predicted concentration established by BBD (glucose 37.43 g/L, yeast extract 0.95 g/L, KH2PO4 1.82 g/L, and MgSO4.7H2O 4.51 g/L), L-Met titer was increased to 3.04 g/L from less than 2.0 g/L. For further enhancement of L-Met biosynthesis, the fermentation conditions of batch cultivation carried out in a 5-L fermentor were optimized, and the optimum results were obtained at an agitation rate of 300 rpm, medium pH of 7.0, and induction temperature of 28 °C. Based on the optimization parameters, fed-batch fermentation with the modified medium was conducted. As a result, great improvement of L-Met titer (12.80 g/L) and yield (0.13 mol/mol) were achieved, with an increase of 38.53% and 30.0% compared with those of the basal medium, respectively. Furthermore, higher L-Met productivity of 0.261 g/L/h was obtained, representing 2.13-fold higher in comparison to the original medium. The results may provide a helpful reference for further study on strain improvement and fermentation control.
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Acknowledgements
The authors gratefully acknowledge the financial support of the National Natural Science Foundation of China (No. 31500031), the Natural Science Foundation of Zhejiang Province (No. LQ14B060004), and the Natural Science Foundation of Zhejiang University of Technology (No. 2012XY010).
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Zhou, HY., Wu, WJ., Niu, K. et al. Enhanced L-methionine production by genetically engineered Escherichia coli through fermentation optimization. 3 Biotech 9, 96 (2019). https://doi.org/10.1007/s13205-019-1609-8
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DOI: https://doi.org/10.1007/s13205-019-1609-8