6LG2
VanR bound to Vanillate
Summary for 6LG2
| Entry DOI | 10.2210/pdb6lg2/pdb |
| Descriptor | Predicted transcriptional regulators, 4-HYDROXY-3-METHOXYBENZOATE (3 entities in total) |
| Functional Keywords | vanr transcription factor vanillate, transcription |
| Biological source | Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025) |
| Total number of polymer chains | 2 |
| Total formula weight | 43519.34 |
| Authors | He, Y.,Bharath, S.R.,Song, H. (deposition date: 2019-12-04, release date: 2020-02-12, Last modification date: 2023-11-22) |
| Primary citation | 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 Cited by 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: 32251291DOI: 10.1038/s41467-020-14918-5 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.6 Å) |
Structure validation
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