Molecular identification and functional characterization of cytochrome P450 monooxygenases from the brown-rot basidiomycete Postia placenta

2011 ◽  
Vol 194 (4) ◽  
pp. 243-253 ◽  
Author(s):  
Masamichi Ide ◽  
Hirofumi Ichinose ◽  
Hiroyuki Wariishi
Keyword(s):  
Cytochrome P450 ◽  
Molecular Identification ◽  
Functional Characterization ◽  
Brown Rot ◽  
Cytochrome P450 Monooxygenases ◽  
Postia Placenta ◽  
P450 Monooxygenases

Functional characterization of cytochrome P450 monooxygenases in the cereal head blight fungusFusarium graminearum

2017 ◽  
Vol 19 (5) ◽  
pp. 2053-2067 ◽  
Author(s):  
Ji Young Shin ◽  
Duc-Cuong Bui ◽  
Yoonji Lee ◽  
Hyejin Nam ◽  
Soyun Jung ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Functional Characterization ◽  
Cytochrome P450 Monooxygenases ◽  
Head Blight ◽  
P450 Monooxygenases

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.

Characterization of cytochrome P450-monooxygenases of Chinese hamsters with respect to aflatoxin B1 activation

Toxicology ◽  
1994 ◽  
Vol 93 (2-3) ◽  
pp. 165-173 ◽  
Author(s):  
Morio Fukuhara ◽  
Eric Antignac ◽  
Naomi Fukusen ◽  
Kazue Kato ◽  
Masanobu Kimura
Keyword(s):  
Cytochrome P450 ◽  
Aflatoxin B1 ◽  
Cytochrome P450 Monooxygenases ◽  
P450 Monooxygenases ◽  
Chinese Hamsters

scholarly journals Construction and Application of a Functional Library of Cytochrome P450 Monooxygenases from the Filamentous Fungus Aspergillus oryzae

2011 ◽  
Vol 77 (9) ◽  
pp. 3147-3150 ◽  
Author(s):  
K. H. M. Nazmul Hussain Nazir ◽  
Hirofumi Ichinose ◽  
Hiroyuki Wariishi
Keyword(s):  
Cytochrome P450 ◽  
Aspergillus Oryzae ◽  
Cytochrome P450 Monooxygenases ◽  
Functional Screening ◽  
Content Type ◽  
P450 Monooxygenases ◽  
Rapid Characterization ◽  
Catalytic Functions ◽  
First Time

ABSTRACTA functional library of cytochrome P450 monooxygenases fromAspergillus oryzae(AoCYPs) was constructed in which 121 isoforms were coexpressed with yeast NADPH-cytochrome P450 oxidoreductase inSaccharomyces cerevisiae. Using this functional library, novel catalytic functions of AoCYPs, such as catalytic potentials of CYP57B3 against genistein, were elucidated for the first time. Comprehensive functional screening promises rapid characterization of catalytic potentials and utility of AoCYPs.

Cloning, Expression and Characterization of a Monooxygenase P450BM3 from Bacillus megaterium ALA2

2012 ◽  
Vol 518-523 ◽  
pp. 5533-5538
Author(s):  
Ting Wang ◽  
Liang Liang Wang ◽  
Xun Li
Keyword(s):  
Escherichia Coli ◽  
Cytochrome P450 ◽  
Linoleic Acid ◽  
Bacillus Megaterium ◽  
Cytochrome P450 Monooxygenases ◽  
P450 Monooxygenases ◽  
Nadph Oxidation ◽  
First Time ◽  
Optimal Ph

Cytochrome P450 monooxygenases are enzymes which are capable of oxidising saturated and unsaturated substrates. P450BM3 from Bacillus megaterium is one of this family. For the first time, the cyp gene for coding P450BM3 from B. megaterium ALA2 has been cloned and expressed in Escherichia coli. The recombinant enzyme is 120 kDa, containing 1049 aa. The highest activity of purified enzyme is 14.8 U/mg towards palmitic acid by monitoring the NADPH oxidation. The optimal pH and temperature were 9.0 and 40°C. The enzyme has higher activity towards linoleic acid, and 2-Methyl-7-octadecene can also be catalyzed which is a precursor of displar.

scholarly journals Biocatalytic Conversion of Avermectin to 4"-Oxo-Avermectin: Characterization of Biocatalytically Active Bacterial Strains and of Cytochrome P450 Monooxygenase Enzymes and Their Genes

2005 ◽  
Vol 71 (11) ◽  
pp. 6968-6976 ◽  
Author(s):  
Volker Jungmann ◽  
István Molnár ◽  
Philip E. Hammer ◽  
D. Steven Hill ◽  
Ross Zirkle ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cytochrome P450 ◽  
Recombinant Proteins ◽  
Oxidation Reaction ◽  
Cytochrome P450 Monooxygenases ◽  
Enzyme Kinetic ◽  
Bacterial Strains ◽  
P450 Monooxygenase ◽  
P450 Monooxygenases

ABSTRACT 4"-Oxo-avermectin is a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate from the natural product avermectin. Seventeen biocatalytically active Streptomyces strains with the ability to oxidize avermectin to 4"-oxo-avermectin in a regioselective manner have been discovered in a screen of 3,334 microorganisms. The enzymes responsible for this oxidation reaction in these biocatalytically active strains were found to be cytochrome P450 monooxygenases (CYPs) and were termed Ema1 to Ema17. The genes for Ema1 to Ema17 have been cloned, sequenced, and compared to reveal a new subfamily of CYPs. Ema1 to Ema16 have been overexpressed in Escherichia coli and purified as His-tagged recombinant proteins, and their basic enzyme kinetic parameters have been determined.

scholarly journals Molecular cloning and functional characterization of two CYP619 cytochrome P450s involved in biosynthesis of patulin in Aspergillus clavatus

Microbiology ◽  
2009 ◽  
Vol 155 (5) ◽  
pp. 1738-1747 ◽  
Author(s):  
Marie Pierre Artigot ◽  
Nicolas Loiseau ◽  
Joelle Laffitte ◽  
Lina Mas-Reguieg ◽  
Souria Tadrist ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Functional Characterization ◽  
Fungal Species ◽  
Current Data ◽  
Cytochrome P450 Monooxygenases ◽  
Aspergillus Clavatus ◽  
P450 Monooxygenases ◽  
Cytochrome P450 Genes ◽  
Methylsalicylic Acid Synthase ◽  
Hydroxybenzyl Alcohol

Patulin is an acetate-derived tetraketide mycotoxin produced by several fungal species, especially Aspergillus, Penicillium and Byssochlamys species. The health risks due to patulin consumption by humans have led many countries to regulate it in human food. Previous studies have shown the involvement of cytochrome P450 monooxygenases in the hydroxylation of two precursors of patulin, m-cresol and m-hydroxybenzylalcohol. In the present study, two cytochrome P450 genes were identified in the genome sequence of Aspergillus clavatus, a patulin-producing species. Both mRNAs were strongly co-expressed during patulin production. CYP619C2, encoded by the first gene, consists of 529 aa, while the second cytochrome, CYP619C3, consists of 524 aa. The coding sequences were used to perform the heterologous expression of functional enzymes in Saccharomyces cerevisiae. The bioconversion assays showed that CYP619C3 catalysed the hydroxylation of m-cresol to yield m-hydroxybenzyl alcohol. CYP619C2 catalysed the hydroxylation of m-hydroxybenzyl alcohol and m-cresol to gentisyl alcohol and 2,5-dihydroxytoluene (toluquinol), respectively. Except for the last compound, all enzyme products are known precursors of patulin. Taken together, these data strongly suggest the involvement of CYP619C2 and CYP619C3 in the biosynthesis of patulin. CYP619C2 and CYP619C3 are located near to two other genes involved in patulin biosynthesis, namely the 6-methylsalicylic acid synthase (6msas) and isoepoxydon dehydrogenase (idh) genes. The current data associated with an analysis of the sequence of A. clavatus suggest the presence of a cluster of 15 genes involved in patulin biosynthesis.

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