Affiliation:
1. grid.258151.a 0000000107081323 Key Laboratory of Industrial Biotechnology, Ministry of Education Jiangnan University 1800 Lihu Road 214122 Wuxi Jiangsu China
2. grid.258151.a 0000 0001 0708 1323 Synergetic Innovation Center of Food Safety and Nutrition 1800 Lihu Road 214122 Wuxi Jiangsu China
3. grid.258151.a 0000000107081323 School of Biotechnology Jiangnan University 1800 Lihu Road 214122 Wuxi Jiangsu China
Abstract
Abstract
Microbial production of l-phenylalanine (l-Phe) from renewable sources has attracted much attention recently. In the present study, Corynebacterium glutamicum 13032 was rationally engineered to produce l-Phe from inexpensive glucose. First, all the l-Phe biosynthesis pathway genes were investigated and the results demonstrated that in addition to AroF and PheA, the native PpsA, TktA, AroE and AroA, and the heterologous AroL and TyrB were also the key enzymes for L-Phe biosynthesis. Through combinational expression of these key enzymes, the l-Phe production was increased to 6.33 ± 0.13 g l−1 which was about 1.48-fold of that of the parent strain C. glutamicum (pXM-pheAfbr-aroFfbr) (fbr, feedback-inhibition resistance). Furthermore, the production of l-Phe was improved to 9.14 ± 0.21 g l−1 by modifying the glucose and l-Phe transport systems and blocking the acetate and lactate biosynthesis pathways. Eventually, the titer of l-Phe was enhanced to 15.76 ± 0.23 g l−1 with a fed-batch fermentation strategy. To the best of our knowledge, this was the highest value reported in rationally engineered C. glutamicum 13032 strains. The results obtained will also contribute to rational engineering of C. glutamicum for production of other valuable aromatic compounds.
Publisher
Oxford University Press (OUP)
Subject
Applied Microbiology and Biotechnology,Biotechnology,Bioengineering
Cited by
24 articles.
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