5YWL
SsCR_L211H
Summary for 5YWL
| Entry DOI | 10.2210/pdb5ywl/pdb |
| Descriptor | Protein induced by osmotic stress (2 entities in total) |
| Functional Keywords | reductase, asymmetric catalysis, substrate inhibition, oxidoreductase |
| Biological source | Scheffersomyces stipitis CBS 6054 (Yeast) |
| Total number of polymer chains | 1 |
| Total formula weight | 37030.86 |
| Authors | Shang, Y.P.,Chen, Q.,Li, A.T.,Yu, H.L.,Xu, J.H. (deposition date: 2017-11-29, release date: 2019-03-06, Last modification date: 2024-03-27) |
| Primary citation | Shang, Y.P.,Chen, Q.,Li, A.T.,Quan, S.,Xu, J.H.,Yu, H.L. Attenuated substrate inhibition of a haloketone reductase via structure-guided loop engineering. J.Biotechnol., 308:141-147, 2020 Cited by PubMed Abstract: Substrate inhibition of enzymes is one of the main obstacles encountered frequently in industrial biocatalysis. Haloketone reductase SsCR was seriously inhibited by substrate 2,2',4'-trichloroacetophenone. In this study, two essential loops were found that have a relationship with substrate binding by conducting X-ray crystal structure analysis. Three key residues were selected from the tips of the loops and substituted with amino acids with lower hydrophobicity to weaken the hydrophobic interactions that bridge the two loops, resulting in a remarkable reduction of substrate inhibition. Among these variants, L211H showed a significant attenuation of substrate inhibition, with a K of 16 mM, which was 16 times that of the native enzyme. The kinetic parameter k/K of L211H was 3.1 × 10 s mM, showing the comparable catalytic efficiency to that of the wild-type enzyme (WT). At the substrate loading of 100 mM, the space time yield of variant L211H in asymmetric reduction of the haloketone was 3-fold higher than that of the WT. PubMed: 31866427DOI: 10.1016/j.jbiotec.2019.12.011 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.098 Å) |
Structure validation
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