By Xiaoshi Wang
In this thesis, Xiaoshi Wang investigates the functionality and mechanism of a newly came across heme-thiolate peroxygenase, AaeAPO. This enzyme classification comes from Agrocybe aegerita and is utilized in the conversion of inert hydrocarbons to alcohols. Xiaoshi's paintings makes a speciality of an extracellular P450 enzyme which isn't constrained in its balance and shortage of solubility and for this reason is correct for common commercial use. the writer demonstrates that the peroxygenase catalyzes quite a lot of reactions. every so often the writer even describes very tough changes in molecules which are hugely inert. Her certain investigations offer a mechanistic framework for a way the peroxygenase catalyzes this sort of huge variety of reactions. an immense spotlight of this thesis is the identity of key short-lived intermediates within the catalytic cycle of the peroxygenase, utilizing speedy kinetic and spectroscopic equipment, in addition to the elucidation of the thermodynamic houses of those high-energy intermediates. This paintings provides new perception into a major category of enzymes.
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Extra resources for A Novel Heme-Thiolate Peroxygenase AaeAPO and Its Implications for C-H Activation Chemistry
3 Left GC trace of product mixture after the reaction of neopentane with H2O2 catalyzed by AaeAPO. Right1H NMR of product mixture and compare it with control. There are new peaks corresponding to neopentanol. The total turnover was about 30. The ﬁnal concentration of ethanol was 13 μM calculated from an internal standard which was added before doing NMR 2 Efﬁcient and Selective Alkane Hydroxylation … 30 Fig. 4 1H NMR of ethane oxidation catalyzed by AaeAPO. New peaks are formed and corresponding to ethanol.
A catalytic amount of flavin analogs were added into the system. Upon initiation by light, with the presence of electron sources such as EDTA or oxalate, flavin got reduced. The reduced FlredH2 reacts with O2 in the solution, produces H2O2 and regenerates the oxidative form of Flox. 6). Control experiments without flavin, electron sources or under the dark results in no products formation. Without AaeAPO, there is also no products formation, flavin only catalyzed by itself. The region- and stereo-selectivity are the same as adding external oxidants.
Acad. Sci. USA 86, 7823–7827 (1989) 25. : Studies in the biochemistry of micro-organisms 65. (A) A survey of chlorine metabolism by moulds (B) Caldariomycin, C5H802Cl2, a metabolic product of Caldariomyces fumago woronichin. Biochem. J. 34, 664–677 (1940) 26. : Chloroperoxidase. I. Isolation and properties of the crystalline glycoprotein. J. Biol. Chem. 241, 1763–1768 (1966) 27. : Chloroperoxidase-catalyzed asymmetric oxidationssubstrate-speciﬁcity and mechanistic study. J. Am. Chem. Soc. 117, 10419–10424 (1995) 28.
A Novel Heme-Thiolate Peroxygenase AaeAPO and Its Implications for C-H Activation Chemistry by Xiaoshi Wang