8WVB

Crystal structure of Lsd18 mutant S195M

Summary for 8WVB

• Entry DOI: 10.2210/pdb8wvb/pdb
• Descriptor: Putative epoxidase LasC, FLAVIN-ADENINE DINUCLEOTIDE, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: fad dependent monooxygenase, epoxidase, mutant, flavoprotein
• Biological source: Streptomyces lasalocidi
• Total number of polymer chains: 2
• Total formula weight: 107023.63

Authors

Liu, N.,Xiao, H.L.,Chen, X. (deposition date: 2023-10-23, release date: 2023-12-13, Last modification date: 2023-12-27)

Primary citation

Liu, N.,Xiao, H.,Zang, Y.,Zhou, L.,Mencius, J.,Yang, Z.,Quan, S.,Chen, X.
Simultaneous Improvement in the Thermostability and Catalytic Activity of Epoxidase Lsd18 for the Synthesis of Lasalocid A.
Int J Mol Sci, 24:-, 2023

PubMed Abstract: Enzymes used in the synthesis of natural products are potent catalysts, capable of efficient and stereoselective chemical transformations. Lsd18 catalyzes two sequential epoxidations during the biosynthesis of lasalocid A, a polyether polyketide natural product. We performed protein engineering on Lsd18 to improve its thermostability and catalytic activity. Utilizing structure-guided methods of FoldX and Rosetta-ddG, we designed 15 mutants of Lsd18. Screening of these mutants using thermal shift assay identified stabilized variants Lsd18-T189M, Lsd18-S195M, and the double mutant Lsd18-T189M-S195M. Trypsin digestion, molecular dynamic simulation, circular dichroism (CD) spectroscopy, and X-ray crystallography provided insights into the molecular basis for the improved enzyme properties. Notably, enhanced hydrophobic interaction within the enzyme core and interaction of the protein with the FAD cofactor appear to be responsible for its better thermostability.

PubMed: 38069118
DOI: 10.3390/ijms242316795

Experimental method

X-RAY DIFFRACTION (2.5 Å)