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KMID : 0545120190290060923
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Journal of Microbiology and Biotechnology
2019 Volume.29 No. 6 p.923 ~ p.932
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Application of Dynamic Regulation to Increase L-Phenylalanine Production in Escherichia coli
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Wu Jie
Liu Yongfei
Zhao Sheng
Sun Jibin
Jin Zhaoxia
Zhang Dawei
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Abstract
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Current strategies of strain improvement processes are mainly focused on enhancing the synthetic pathways of the products. However, excessive metabolic flux often creates metabolic imbalances, which lead to growth retardation and ultimately limit the yield of the product. To solve this problem, we applied a dynamic regulation strategy to produce L-phenylalanine (LPhe) in Escherichia coli. First, we constructed a series of Phe-induced promoters that exhibited different strengths through modification of the promoter region of tyrP. Then, two engineered promoters were separately introduced into a Phe-producing strain xllp1 to dynamically control the expression level of one pathway enzyme AroK. Batch fermentation results of the strain xllp3 showed that the titer of Phe reached 61.3 g/l at 48 h, representing a titer of 1.36- fold of the strain xllp1 (45.0 g/l). Moreover, the L-Phe yields on glucose of xllp3 (0.22 g/g) were also greatly improved, with an increase of 1.22-fold in comparison with the xllp1 (0.18 g/ g). In summary, we successfully improved the titer of Phe by using dynamic regulation of one key enzyme and this strategy can be applied for improving the performance of strains producing other aromatic amino acids and derived compounds.
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KEYWORD
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L-Phenylalanine, E. coli, dynamic regulation, AroK, fermentation
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