Tunnel engineering for modulating the substrate preference in cytochrome P450_BsβHI.

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, P450_BsβHI (Q85H/V170I) derived from hydroxylase P450_Bsβ 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 P450_BsβHI. 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 p

SUBMITTER: Meng S 

PROVIDER: S-EPMC10992877 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature