000131693 001__ 131693
000131693 005__ 20240219150720.0
000131693 0247_ $$2doi$$a10.1128/JB.00286-06
000131693 0248_ $$2sideral$$a90564
000131693 037__ $$aART-2006-90564
000131693 041__ $$aeng
000131693 100__ $$aFunhoff, E. G.
000131693 245__ $$aCYP153A6, a Soluble P450 Oxygenase Catalyzing Terminal-Alkane Hydroxylation
000131693 260__ $$c2006
000131693 5203_ $$aThe first and key step in alkane metabolism is the terminal hydroxylation of alkanes to 1-alkanols, a reaction catalyzed by a family of integral-membrane diiron enzymes related to Pseudomonas putida GPo1 AlkB, by a diverse group of methane, propane, and butane monooxygenases and by some membrane-bound cytochrome P450s. Recently, a family of cytoplasmic P450 enzymes was identified in prokaryotes that allow their host to grow on aliphatic alkanes. One member of this family, CYP153A6 from Mycobacterium sp. HXN-1500, hydroxylates medium-chain-length alkanes (C6 to C11) to 1-alkanols with a maximal turnover number of 70 min−1 and has a regiospecificity of ≥95% for the terminal carbon atom position. Spectroscopic binding studies showed that C6-to-C11 aliphatic alkanes bind in the active site with Kd values varying from ∼20 nM to 3.7 μM. Longer alkanes bind more strongly than shorter alkanes, while the introduction of sterically hindering groups reduces the affinity. This suggests that the substrate-binding pocket is shaped such that linear alkanes are preferred. Electron paramagnetic resonance spectroscopy in the presence of the substrate showed the formation of an enzyme-substrate complex, which confirmed the binding of substrates observed in optical titrations. To rationalize the experimental observations on a molecular scale, homology modeling of CYP153A6 and docking of substrates were used to provide the first insight into structural features required for terminal alkane hydroxylation.
000131693 540__ $$9info:eu-repo/semantics/openAccess$$aAll rights reserved$$uhttp://www.europeana.eu/rights/rr-f/
000131693 590__ $$a3.993$$b2006
000131693 591__ $$aMICROBIOLOGY$$b16 / 88 = 0.182$$c2006$$dQ1$$eT1
000131693 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000131693 700__ $$aBauer, U.
000131693 700__ $$0(orcid)0000-0002-1827-1250$$aGarcia Rubio, Ines
000131693 700__ $$aWitholt, B.
000131693 700__ $$avan Beilen, J. B.
000131693 773__ $$g188, 14 (2006), 5220-5227$$pJ. bacteriol.$$tJOURNAL OF BACTERIOLOGY$$x0021-9193
000131693 8564_ $$s584474$$uhttps://zaguan.unizar.es/record/131693/files/texto_completo.pdf$$yVersión publicada
000131693 8564_ $$s3388731$$uhttps://zaguan.unizar.es/record/131693/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000131693 909CO $$ooai:zaguan.unizar.es:131693$$particulos$$pdriver
000131693 951__ $$a2024-02-19-13:23:49
000131693 980__ $$aARTICLE