Open AccessConstruction and application of novel feedback‐resistant 3‐deoxy‐ d ‐arabino‐heptulosonate‐7‐phosphate synthases by engineering the N ‐terminal domain for l ‐phenylalanine synthesis
Author(s) -
Zhang Chuanzhi,
Kang Zhen,
Zhang Junli,
Du Guocheng,
Chen Jian,
Yu Xiaobin
Publication year - 2014
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12397
Subject(s) - phosphate , terminal (telecommunication) , bioorganic chemistry , phenylalanine , biochemistry , chemistry , protein engineering , stereochemistry , biology , enzyme , amino acid , computer science , telecommunications
3‐Deoxy‐ d ‐arabino‐heptulosonate 7‐phosphate synthase (DAHP synthase) encoded by aro F is the first enzyme of the shikimate pathway. In the present study, an A ro F variant with a deficiency in residue I le11 (named A ro F *) was shown to be insensitive to l ‐tyrosine. According to three‐dimensional structure analysis, nine A ro F variants were constructed with truncation of different N‐terminal fragments, and overexpression of the variants A ro F Δ(1–9) , A ro F Δ(1–10) , A ro F Δ(1–12) and, in particular, A ro F Δ(1–11) significantly increased the accumulation of l ‐phenylalanine ( l ‐Phe). However, the A ro G and A ro H variants with similar truncations of the N‐terminal fragments decreased the production of l ‐ P he. By co‐overexpressing A ro F Δ(1–11) and P he A fbr , the production of l ‐Phe was increased from 2.36 ± 0.07 g L −1 (co‐overexpression of the wild‐type A ro F and P he A fbr ) to 4.29 ± 0.06 g L −1 . The novel variant A ro F Δ(1–11) showed great potential for the production of aromatic amino acids and their derivatives.