Structural and Biochemical Characterization of the Cytochrome P450 CypX (CYP134A1) fromBacillus subtilis: A Cyclo-l-leucyl-l-leucyl Dipeptide Oxidase

Biochemistry ◽  
2010 ◽  
Vol 49 (34) ◽  
pp. 7282-7296 ◽  
Author(s):  
Max J. Cryle ◽  
Stephen G. Bell ◽  
Ilme Schlichting
Keyword(s):  
Cytochrome P450 ◽  
Biochemical Characterization

scholarly journals Identification and characterization of cytochrome P450 1232A24 and 1232F1 from Arthrobacter sp. and their role in the metabolic pathway of papaverine

2019 ◽  
Vol 166 (1) ◽  
pp. 51-66 ◽  
Author(s):  
Jan M Klenk ◽  
Max-Philipp Fischer ◽  
Paulina Dubiel ◽  
Mahima Sharma ◽  
Benjamin Rowlinson ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Metabolic Pathway ◽  
Biochemical Characterization ◽  
Gene Clusters ◽  
Cytochrome P450 Monooxygenases ◽  
Dihydroxyphenylacetic Acid ◽  
Meta Position ◽  
Redox Partners

AbstractCytochrome P450 monooxygenases (P450s) play crucial roles in the cell metabolism and provide an unsurpassed diversity of catalysed reactions. Here, we report the identification and biochemical characterization of two P450s from Arthrobacter sp., a Gram-positive organism known to degrade the opium alkaloid papaverine. Combining phylogenetic and genomic analysis suggested physiological roles for P450s in metabolism and revealed potential gene clusters with redox partners facilitating the reconstitution of the P450 activities in vitro. CYP1232F1 catalyses the para demethylation of 3,4-dimethoxyphenylacetic acid to homovanillic acid while CYP1232A24 continues demethylation to 3,4-dihydroxyphenylacetic acid. Interestingly, the latter enzyme is also able to perform both demethylation steps with preference for the meta position. The crystal structure of CYP1232A24, which shares only 29% identity to previous published structures of P450s helped to rationalize the preferred demethylation specificity for the meta position and also the broader substrate specificity profile. In addition to the detailed characterization of the two P450s using their physiological redox partners, we report the construction of a highly active whole-cell Escherichia coli biocatalyst expressing CYP1232A24, which formed up to 1.77 g l−1 3,4-dihydroxyphenylacetic acid. Our results revealed the P450s’ role in the metabolic pathway of papaverine enabling further investigation and application of these biocatalysts.

Biochemical Characterization of the Human Liver Cytochrome P450 Arachidonic Acid Epoxygenase Pathway

1996 ◽  
Vol 330 (1) ◽  
pp. 87-96 ◽  
Author(s):  
Darryl C. Zeldin ◽  
Cindy R. Moomaw ◽  
Nate Jesse ◽  
Kenneth B. Tomer ◽  
Jeffrey Beetham ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Cytochrome P450 ◽  
Human Liver ◽  
Biochemical Characterization ◽  
Liver Cytochrome

Biochemical Characterization of the Cytochrome P450 CYP107CB2 from Bacillus lehensis G1

2018 ◽  
Vol 37 (2) ◽  
pp. 180-193
Author(s):  
Swi See Ang ◽  
Abu Bakar Salleh ◽  
Leow Thean Chor ◽  
Yahaya M. Normi ◽  
Bimo Ario Tejo ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Biochemical Characterization

Identification and characterization of a novel class of self-sufficient cytochrome P450 hydroxylase involved in cyclohexanecarboxylate degradation in Paraburkholderia terrae strain KU-64

2021 ◽  
Author(s):  
Taisei Yamamoto ◽  
Yoshie Hasegawa ◽  
Hiroaki Iwaki
Keyword(s):  
Cytochrome P450 ◽  
Biochemical Characterization ◽  
Functional Characterization ◽  
Physiological Role ◽  
Degradation Pathway ◽  
Cytochrome P450 Monooxygenases ◽  
Recombinant Protein Purification ◽  
Cytochrome P450 Hydroxylase ◽  
P450 Monooxygenases

ABSTRACT Cytochrome P450 monooxygenases play important roles in metabolism. Here, we report the identification and biochemical characterization of P450CHC, a novel self-sufficient cytochrome P450, from cyclohexanecarboxylate-degrading Paraburkholderia terrae KU-64. P450CHC was found to comprise a [2Fe-2S] ferredoxin domain, NAD(P)H-dependent FAD-containing reductase domain, FCD domain, and cytochrome P450 domain (in that order from the N terminus). Reverse transcription–polymerase chain reaction results indicated that the P450CHC-encoding chcA gene was inducible by cyclohexanecarboxylate. chcA overexpression in Escherichia coli and recombinant protein purification enabled functional characterization of P450CHC as a catalytically self-sufficient cytochrome P450 that hydroxylates cyclohexanecarboxylate. Kinetic analysis indicated that P450CHC largely preferred NADH (Km = 0.011 m m) over NADPH (Km = 0.21 m m). The Kd, Km, and kcat values for cyclohexanecarboxylate were 0.083 m m, 0.084 m m, and 15.9 s−1, respectively. The genetic and biochemical analyses indicated that the physiological role of P450CHC is initial hydroxylation in the cyclohexanecarboxylate degradation pathway.

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