A Cold-Active Flavin-Dependent Monooxygenase from Janthinobacterium svalbardensis Unlocks Applications of Baeyer-Villiger Monooxygenases at Low Temperature

Simple item page
dc.contributor.authorChanique, Andrea M.
dc.contributor.authorPolidori, Nakia
dc.contributor.authorSovic, Lucija
dc.contributor.authorKracher, Daniel
dc.contributor.authorAssil-Companioni, Leen
dc.contributor.authorGaluska, Philipp
dc.contributor.authorParra, Loreto P.
dc.contributor.authorGruber, Karl
dc.contributor.authorKourist, Robert
dc.date.accessioned2025-01-20T20:07:04Z
dc.date.available2025-01-20T20:07:04Z
dc.date.issued2023
dc.description.abstractCold-active enzymes maintain a large part of their optimal activity at low temperatures. Therefore, they can be used to avoid side reactions and preserve heat sensitive compounds. Baeyer-Villiger monooxygenases (BVMO) utilize molecular oxygen as a co-substrate to catalyze reactions widely employed for steroid, agrochemical, antibiotic, and pheromone production. Oxygen has been described as the rate-limiting factor for some BVMO applications, thereby hindering their efficient utilization. Considering that oxygen solubility in water increases by 40% when the temperature is decreased from 30 to 10 degrees C, we set out to identify and characterize a cold-active BVMO. Using genome mining in the Antarctic organism Janthinobacterium svalbardensis, a cold active type II flavin-dependent monooxygenase (FMO) was discovered. The enzyme shows promiscuity toward NADH and NADPH and high activity between 5 and 25 degrees C. The enzyme catalyzes the monooxygenation and sulfoxidation of a wide range of ketones and thioesters. The high enantioselectivity in the oxidation of norcamphor (eeS = 56%, eeP > 99%, E > 200) demonstrates that the generally higher flexibility observed in the active sites of cold-active enzymes, which compensates for the lower motion at cold temperatures, does not necessarily reduce the selectivity of these enzymes. To gain a better understanding of the unique mechanistic features of type II FMOs, we determined the structure of the dimeric enzyme at 2.5 angstrom resolution. While the unusual N-terminal domain has been related to the catalytic properties of type II FMOs, the structure shows a SnoaL-like N-terminal domain that is not interacting directly with the active site. The active site of the enzyme is accessible only through a tunnel, with Tyr-458, Asp-217, and His-216 as catalytic residues, a combination not observed before in FMOs and BVMOs.
dc.fuente.origenWOS
dc.identifier.doi10.1021/acscatal.2c05160
dc.identifier.issn2155-5435
dc.identifier.urihttps://doi.org/10.1021/acscatal.2c05160
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/91761
dc.identifier.wosidWOS:000939553800001
dc.issue.numero6
dc.language.isoen
dc.pagina.final3562
dc.pagina.inicio3549
dc.revistaAcs catalysis
dc.rightsacceso restringido
dc.subjectbiocatalysis
dc.subjectenantioselectivity
dc.subjectcold-active
dc.subjectBaeyer-Villiger monooxygenase
dc.subjectcofactor promiscuity
dc.titleA Cold-Active Flavin-Dependent Monooxygenase from Janthinobacterium svalbardensis Unlocks Applications of Baeyer-Villiger Monooxygenases at Low Temperature
dc.typeartículo
dc.volumen13
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
Files
Collections
Artículos de revistas