title: Construction and application of novel feedback-resistant 3-deoxy-d-arabino-heptulosonate-7-phosphate synthases by engineering the N-terminal domain for L-phenylalanine synthesis.
abstract: 3-Deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAHP synthase) encoded by aroF is the first enzyme of the shikimate pathway. In the present study, an AroF variant with a deficiency in residue Ile11 (named AroF*) was shown to be insensitive to l-tyrosine. According to three-dimensional structure analysis, nine AroF variants were constructed with truncation of different N-terminal fragments, and overexpression of the variants AroF(Δ(1-9)) , AroF(Δ(1-10)) , AroF(Δ(1-12)) and, in particular, AroF(Δ(1-11)) significantly increased the accumulation of l-phenylalanine (l-Phe). However, the AroG and AroH variants with similar truncations of the N-terminal fragments decreased the production of l-Phe. By co-overexpressing AroF(Δ(1-11)) and PheA(fbr) , the production of l-Phe was increased from 2.36 ± 0.07 g L(-1) (co-overexpression of the wild-type AroF and PheA(fbr) ) to 4.29 ± 0.06 g L(-1) . The novel variant AroF(Δ(1-11)) showed great potential for the production of aromatic amino acids and their derivatives.
authors: Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, China; The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, China; School of Biotechnology, Jiangnan University, Wuxi, China.
authors: Chuanzhi
authors: C
authors: Zhang
authors: Zhen
authors: Z
authors: Kang
authors: Junli
authors: J
authors: Zhang
authors: Guocheng
authors: G
authors: Du
authors: Jian
authors: J
authors: Chen
authors: Xiaobin
authors: X
authors: Yu
mesh_terms: D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000235:genetics / Q000378:metabolism*; D003352:Corynebacterium / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D010649:Phenylalanine / Q000096:biosynthesis*; D015202:Protein Engineering / Q000379:methods*; D017433:Protein Structure, Secondary
chemical_list: D007527:Isoenzymes; D010649:Phenylalanine; D010723:3-Deoxy-7-Phosphoheptulonate Synthase
keywords: AroF; Corynebacterium glutamicum; DAHP synthases; l-phenylalanine