By Xiaoshi Wang
In this thesis, Xiaoshi Wang investigates the functionality and mechanism of a newly chanced on heme-thiolate peroxygenase, AaeAPO. This enzyme category 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 restricted in its balance and shortage of solubility and for this reason is proper for common business use. the writer demonstrates that the peroxygenase catalyzes quite a lot of reactions. occasionally the writer even describes very tricky changes in molecules which are hugely inert. Her specific investigations offer a mechanistic framework for the way the peroxygenase catalyzes the sort of huge variety of reactions. an enormous spotlight of this thesis is the id of key short-lived intermediates within the catalytic cycle of the peroxygenase, utilizing swift 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 huge classification of enzymes.
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Extra resources for A Novel Heme-Thiolate Peroxygenase AaeAPO and Its Implications for C-H Activation Chemistry
Early studies have showed that among all of metabolites of ticlopidine, 2-oxo-ticlopidine was the only metabolite that had anti-platelet activity in vivo . Our HPLC-MS analysis showed that 40 % of the total metabolites by AaeAPO in vitro was 2-oxo-ticlopidine (Fig. 5). 4 Several examples of drug metabolites by AaeAPO 32 2 Efﬁcient and Selective Alkane Hydroxylation … Fig. 6 Water suppressed 1H NMR of ibuprofen metabolites time and ESI-MS was conﬁrmed by the authentic standard. So AaeAPO might have the same function as CYP2C19 in the liver .
For example, AaeAPO prefers tertiary and secondary carbons but not primary carbons. Most linear alkanes (C2–C16) hydroxylations happened at the 2-position and 3-position. 1 Experiments were performed by Dr. Sebastian Peter and data were from Ref.  a Product distribution determined as ratio of a speciﬁc alcohol product in relation to the total amount of all alcohol products b Product distribution for ketones was similar to that of alcohol product distribution. The numbers reported here are the total of all ketones (%) relative to total products (alcohols and ketones) complete stereoselectivity for the (R)-enantiomer of the corresponding 3-alcohol.
For example, CPO catalyzes C–H hydroxylation at benzylic position while CYPs are able to catalyze aliphatic hydroxylation with stronger C–H bonds (*101 kcal/mol). Because of its good solubility, easy handling and catalytic versatility, CPO has become a good model for the study of P450 reaction mechanisms . It has also been utilized for synthetic and biocatalytic applications. The active site of CPO includes a glutamic acid and a histidine as the acidic and basic catalytic residues  (Fig.