title: Controlled oxidation of aliphatic CH bonds in metallo-monooxygenases: mechanistic insights derived from studies on deuterated and fluorinated hydrocarbons.
abstract: The control over the regio- and/or stereo-selective aliphatic CH oxidation by metalloenzymes is of great interest to scientists. Typically, these enzymes invoke host-guest chemistry to sequester the substrates within the protein pockets, exploiting sizes, shapes and specific interactions such as hydrogen-bonding, electrostatic forces and/or van der Waals interactions to control the substrate specificity, regio-specificity and stereo-selectivity. Over the years, we have developed a series of deuterated and fluorinated variants of these hydrocarbon substrates as probes to gain insights into the controlled CH oxidations of hydrocarbons facilitated by these enzymes. In this review, we illustrate the application of these designed probes in the study of three monooxygenases: (i) the particulate methane monooxygenase (pMMO) from Methylococcus capsulatus (Bath), which oxidizes straight-chain C1-C5 alkanes and alkenes to form their corresponding 2-alcohols and epoxides, respectively; (ii) the recombinant alkane hydroxylase (AlkB) from Pseudomonas putida GPo1, which oxidizes the primary CH bonds of C5-C12 linear alkanes; and (iii) the recombinant cytochrome P450 from Bacillus megaterium, which oxidizes C12-C20 fatty acids at the ω-1, ω-2 or ω-3 CH positions.
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan; Institute of Fisheries Science, National Taiwan University, Taipei 106, Taiwan; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yao-Sheng
authors.initials: YS
authors.lastname: Chen
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Wen-I
authors.initials: WI
authors.lastname: Luo
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan; Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei 106, Taiwan.
authors.forename: Chung-Ling
authors.initials: CL
authors.lastname: Yang
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yi-Jung
authors.initials: YJ
authors.lastname: Tu
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Chun-Wei
authors.initials: CW
authors.lastname: Chang
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan; Department of Chemistry, National Cheng Kung University, Tainan 701, Taiwan.
authors.forename: Chih-Hsiang
authors.initials: CH
authors.lastname: Chiang
authors.affiliation: Institute of Fisheries Science, National Taiwan University, Taipei 106, Taiwan; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Chi-Yao
authors.initials: CY
authors.lastname: Chang
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Sunney I
authors.initials: SI
authors.lastname: Chan
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan; Department of Chemistry, National Cheng Kung University, Tainan 701, Taiwan. Electronic address: sfyu@gate.sinica.edu.tw.
authors.forename: Steve S-F
authors.initials: SS
authors.lastname: Yu
mesh_terms: D001410:Bacillus megaterium / Q000737:chemistry / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D042926:Cytochrome P-450 CYP4A / Q000737:chemistry* / Q000302:isolation & purification; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D003903:Deuterium / Q000737:chemistry*; D006845:Hydrocarbons, Fluorinated / Q000737:chemistry*; D007700:Kinetics; D020619:Methylococcus capsulatus / Q000737:chemistry / Q000201:enzymology; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000737:chemistry* / Q000302:isolation & purification; D016958:Pseudomonas putida / Q000737:chemistry / Q000201:enzymology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D006845:Hydrocarbons, Fluorinated; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D003903:Deuterium; D010105:Oxygenases; C021555:methane monooxygenase; D042926:Cytochrome P-450 CYP4A
keywords: CH activation; Enzyme catalysis; Fluorinated substituents; Isotopomers; Metalloprotein; Monooxygenase