5yw4

Structure-Guided Engineering of Reductase: Efficient Attenuating Substrate Inhibition in Asymmetric Catalysis

spinqualitylabelsall models 5yw4, resolution 2.05Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5yw4 is a 1 chain structure with sequence from Scheffersomyces stipitis CBS 6054. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 2.047Å Ligands: NAP Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function A3LWG4_PICST Publication Abstract from PubMed 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 Ki of 16 mM, which was 16 times that of the native enzyme. The kinetic parameter kcat/Km of L211H was 3.1 x 10(3) s(-1) mM(-1), 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. Attenuated substrate inhibition of a haloketone reductase via structure-guided loop engineering.,Shang YP, Chen Q, Li AT, Quan S, Xu JH, Yu HL J Biotechnol. 2020 Jan 20;308:141-147. doi: 10.1016/j.jbiotec.2019.12.011. Epub, 2019 Dec 19. PMID:31866427 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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