Abstract
l-Phenylalanine is an important amino acid commercially, and therefore optimization of its manufacture is of interest. We constructed a range of mutant alleles of AroG, the enzyme involved in the first step of phenylalanine biosynthesis. Three single-site mutant alleles were constructed (aroG8, aroG15, and aroG29), which were then combined to generate three double-site aroG fbr mutant alleles (aroG8/15, aroG8/29, and aroG15/29). Enzymatic activity, feedback inhibition, and fermentation were analyzed in all of the mutants. All double-site mutants, except AroG15/29, showed higher enzymatic activity and greater resistance to feedback inhibition than their respective single-site mutants. The E. coli strain carrying the aroG8/15 allele produced a phenylalanine titer of 26.78 g/l, a 116 % improvement over the control phenylalanine overproducing strain (12.41 g/l). Our findings provide an effective method for modifying phenylalanine biosynthetic genes, which may be applied to optimize the commercial manufacture of phenylalanine.
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Acknowledgments
We thank Ping Qin and Songyi Cao (Biological Science and Technology College, Shenyang Agricultural University, China) for their technical assistance throughout the study. This work was supported by the National Natural Science Foundation of China (Nos. 31370283, 30870190, and 31000673), and by the Major Program of Science and Technique Foundation from the Technological Office of Liaoning (No. 2008208001).
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Ding, R., Liu, L., Chen, X. et al. Introduction of two mutations into AroG increases phenylalanine production in Escherichia coli . Biotechnol Lett 36, 2103–2108 (2014). https://doi.org/10.1007/s10529-014-1584-4
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DOI: https://doi.org/10.1007/s10529-014-1584-4