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. Issue 1 (5th March 2014)

• Record Type: Journal Article
• Authors: Zhang, Chuanzhi; Kang, Zhen; Zhang, Junli; Du, Guocheng; Chen, Jian; Yu, Xiaobin
• Is Part Of: FEMS microbiology letters. Volume 353:Issue 1(2014:Apr.)
• Page Start: 11
• Page End: 18
• Publication Date: 2014-03-05
• Subjects: Microbiology -- Periodicals; 579
• DOI: 10.1111/1574-6968.12397
• Languages: English
• ISSNs: 0378-1097

Abstract

<abstract abstract-type="main" id="fml12397-abs-0001"> <title>Abstract</title> <p>3‐Deoxy‐<sc>d</sc>‐arabino‐heptulosonate 7‐phosphate synthase (DAHP synthase) encoded by <italic>aroF</italic> 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 <sc>l</sc>‐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 <sc>l</sc>‐phenylalanine (<sc>l</sc>‐Phe). However, the AroG and AroH variants with similar truncations of the N‐terminal fragments decreased the production of <sc>l</sc>‐Phe. By co‐overexpressing AroF^Δ(1–11) and PheA^fbr, the production of <sc>l</sc>‐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.</p> </abstract>