AI Article Synopsis
- Researchers fused genes from cytochrome P450 BM3 and formate dehydrogenase to create multifunctional enzymes aimed at improving stability and activity.
- The study varied the arrangement of the genes and linkers and compared the fused enzymes with individual ones in terms of substrate conversion and stability.
- A significant increase in activity (up to threefold) was noted for the fusion constructs, indicating that the fusion may cause conformational changes in P450 BM3, although no NADPH channeling was observed.
Article Abstract
Fusion of multiple enzymes to multifunctional constructs has been recognized as a viable strategy to improve enzymatic properties at various levels such as stability, activity and handling. In this study, the genes coding for cytochrome P450 BM3 from B. megaterium and formate dehydrogenase from Pseudomonas sp. were fused to enable both substrate oxidation catalyzed by P450 BM3 and continuous cofactor regeneration by formate dehydrogenase within one construct. The order of the genes in the fusion as well as the linkers that bridge the enzymes were varied. The resulting constructs were compared to individual enzymes regarding substrate conversion, stability and kinetic parameters to examine whether fusion led to any substantial improvements of enzymatic properties. Most noticeably, an activity increase of up to threefold was observed for the fusion constructs with various substrates which were partly attributed to the increased diflavin reductase activity of the P450 BM3. We suggest that P450 BM3 undergoes conformational changes upon fusion which resulted in altered properties, however, no NADPH channeling was detected for the fusion constructs.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8568981 | PMC |
| http://dx.doi.org/10.1038/s41598-021-00957-5 | DOI Listing |
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