共表达 pheA(fbr) 和 aroF(wt) 增强 l-苯丙氨酸生物合成。
Enhanced l-phenylalanine biosynthesis by co-expression of pheA(fbr) and aroF(wt).
机构信息
Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122 Jiangsu, China.
出版信息
Bioresour Technol. 2010 Jun;101(11):4151-6. doi: 10.1016/j.biortech.2010.01.043.
A beta-2-thienylalanine-resistant E. coli K12 mutant carrying a Thr326Pro mutation in the regulation (R) domain of pheA (pheA(fbr)) was obtained by N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis. In the presence of 200mM l-phenylalanine (l-Phe), a recombinant E. coli WSH-Z06 (pAP-B03) carrying pheA(fbr) as well as wild-type aroF (aroF(wt)) exhibited more than 70% of the chorismate mutase-prephenate dehydratase (CM-PDT) activity as observed in the absence of this amino acid. The l-Phe titer of WSH-Z06 (pAP-B03) reached 35.38g/L in a 3-L fermentor, which was 2.81-fold higher than that of the original strain E. coli WSH-Z06. Furthermore, the l-Phe yield on glucose of WSH-Z06 (pAP-B03) (0.26mol/mol) was twice that of E. coli WSH-Z06. This recombinant E. coli WSH-Z06 (pAP-B03) is a potential strain for over-production of l-Phe.
携带 pheA（pheA(fbr)）调控（R）域 Thr326Pro 突变的β-2-噻吩丙氨酸抗性大肠杆菌 K12 突变体通过 N-甲基-N'-硝基-N-亚硝基胍（NTG）诱变获得。在存在 200mM l-苯丙氨酸（l-Phe）的情况下，携带 pheA(fbr)以及野生型 aroF（aroF(wt)）的重组大肠杆菌 WSH-Z06（pAP-B03）表现出的分支酸变位酶-预苯酸脱水酶（CM-PDT）活性超过了在不存在这种氨基酸的情况下观察到的活性的 70%。WSH-Z06（pAP-B03）在 3L 发酵罐中的 l-Phe 产量达到 35.38g/L，比原始菌株大肠杆菌 WSH-Z06 高 2.81 倍。此外，WSH-Z06（pAP-B03）（0.26mol/mol）的葡萄糖上 l-Phe 的产率是大肠杆菌 WSH-Z06 的两倍。这种重组大肠杆菌 WSH-Z06（pAP-B03）是 l-Phe 过量生产的潜在菌株。
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