6LG2

VanR bound to Vanillate

6LG2 の概要

• エントリーDOI: 10.2210/pdb6lg2/pdb
• 分子名称: Predicted transcriptional regulators, 4-HYDROXY-3-METHOXYBENZOATE (3 entities in total)
• 機能のキーワード: vanr transcription factor vanillate, transcription
• 由来する生物種: Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 43519.34

構造登録者

He, Y.,Bharath, S.R.,Song, H. (登録日: 2019-12-04, 公開日: 2020-02-12, 最終更新日: 2023-11-22)

主引用文献

Yao, J.,He, Y.,Su, N.,Bharath, S.R.,Tao, Y.,Jin, J.M.,Chen, W.,Song, H.,Tang, S.Y.
Developing a highly efficient hydroxytyrosol whole-cell catalyst by de-bottlenecking rate-limiting steps.
Nat Commun, 11:1515-1515, 2020

PubMed Abstract: Hydroxytyrosol is an antioxidant free radical scavenger that is biosynthesized from tyrosine. In metabolic engineering efforts, the use of the mouse tyrosine hydroxylase limits its production. Here, we design an efficient whole-cell catalyst of hydroxytyrosol in Escherichia coli by de-bottlenecking two rate-limiting enzymatic steps. First, we replace the mouse tyrosine hydroxylase by an engineered two-component flavin-dependent monooxygenase HpaBC of E. coli through structure-guided modeling and directed evolution. Next, we elucidate the structure of the Corynebacterium glutamicum VanR regulatory protein complexed with its inducer vanillic acid. By switching its induction specificity from vanillic acid to hydroxytyrosol, VanR is engineered into a hydroxytyrosol biosensor. Then, with this biosensor, we use in vivo-directed evolution to optimize the activity of tyramine oxidase (TYO), the second rate-limiting enzyme in hydroxytyrosol biosynthesis. The final strain reaches a 95% conversion rate of tyrosine. This study demonstrates the effectiveness of sequentially de-bottlenecking rate-limiting steps for whole-cell catalyst development.

PubMed: 32251291
DOI: 10.1038/s41467-020-14918-5

実験手法

X-RAY DIFFRACTION (1.6 Å)