Tunnel engineering for modulating the substrate preference in cytochrome P450BsβHI
Open Access
- 3 April 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Bioresources and Bioprocessing
- Vol. 8 (1), 1-10
- https://doi.org/10.1186/s40643-021-00379-1
Abstract
An active site is normally located inside enzymes, hence substrates should go through a tunnel to access the active site. Tunnel engineering is a powerful strategy for refining the catalytic properties of enzymes. Here, P450BsβHI (Q85H/V170I) derived from hydroxylase P450Bsβ from Bacillus subtilis was chosen as the study model, which is reported as a potential decarboxylase. However, this enzyme showed low decarboxylase activity towards long-chain fatty acids. Here, a tunnel engineering campaign was performed for modulating the substrate preference and improving the decarboxylation activity of P450BsβHI. The finally obtained BsβHI-F79A variant had a 15.2-fold improved conversion for palmitic acid; BsβHI-F173V variant had a 3.9-fold improved conversion for pentadecanoic acid. The study demonstrates how the substrate preference can be modulated by tunnel engineering strategy.Other Versions
Funding Information
- National Natural Science Foundation of China (31961133017, 21978017, 21978020, 21861132017)
- European Union’s Horizon 2020 (870294)
This publication has 45 references indexed in Scilit:
- MOLE 2.0: advanced approach for analysis of biomacromolecular channelsJournal of Cheminformatics, 2013
- Conformational diversity and ligand tunnels of mammalian cytochrome P450sBiotechnology and Applied Biochemistry, 2013
- Modular Synthase-Encoding Gene Involved in α-Olefin Biosynthesis in Synechococcus sp. Strain PCC 7002Applied and Environmental Microbiology, 2011
- Terminal Olefin (1-Alkene) Biosynthesis by a Novel P450 Fatty Acid Decarboxylase from Jeotgalicoccus SpeciesApplied and Environmental Microbiology, 2011
- ConSurf 2010: calculating evolutionary conservation in sequence and structure of proteins and nucleic acidsNucleic Acids Research, 2010
- The gates of ion channels and enzymesTrends in Biochemical Sciences, 2010
- Genes Involved in Long-Chain Alkene Biosynthesis in Micrococcus luteusApplied and Environmental Microbiology, 2010
- GROMACS 4: Algorithms for Highly Efficient, Load-Balanced, and Scalable Molecular SimulationJournal of Chemical Theory and Computation, 2008
- Hydrophobicity and structural classes in proteins"Protein Engineering, Design and Selection", 1992
- The characterization of amino acid sequences in proteins by statistical methodsJournal of Theoretical Biology, 1968