Mechanisms of resistance to thiamethoxam and dinotefuran compared to imidacloprid in the brown planthopper: Roles of cytochrome P450 monooxygenase and a P450 gene CYP6ER1

ABSTRACT The genome sequence of Mycobacterium leprae revealed a single open reading frame, ML2088 (CYP164A1), encoding a putative full-length cytochrome P450 monooxygenase and 12 pseudogenes. We have identified a homolog of ML2088 in Mycobacterium smegmatis and report here the cloning, expression, purification, and azole-binding characteristics of this cytochrome P450 (CYP164A2). CYP164A2 is 1,245 bp long and encodes a protein of 414 amino acids and molecular mass of 45 kDa. CYP164A2 has 60% identity with Mycobacterium leprae CYP161A1 and 66 to 69% identity with eight other mycobacterial CYP164A1 homologs, with three identified highly conserved motifs. Recombinant CYP164A2 has the typical spectral characteristics of a cytochrome P450 monooxygenase, predominantly in the ferric low-spin state. Unusually, the spin state was readily modulated by increasing ionic strength at pH 7.5, with 50% high-spin occupancy achieved with 0.14 M NaCl. CYP164A2 bound clotrimazole, econazole, and miconazole strongly (Kd , 1.2 to 2.5 μM); however, strong binding with itraconazole, ketoconazole, and voriconazole was only observed in the presence of 0.5 M NaCl. Fluconazole did not bind to CYP164A2 at pH 7.5 and no discernible type II binding spectrum was observed.

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Overexpression of a cytochrome P450 monooxygenase, CYP6ER1, is associated with resistance to imidacloprid in the brown planthopper, Nilaparvata lugens

Insect Molecular Biology ◽

10.1111/j.1365-2583.2011.01105.x ◽

2011 ◽

Vol 20 (6) ◽

pp. 763-773 ◽

Cited By ~ 114

Author(s):

C. Bass ◽

R. A. Carvalho ◽

L. Oliphant ◽

A. M. Puinean ◽

L. M. Field ◽

...

Keyword(s):

Cytochrome P450 ◽

Brown Planthopper ◽

Nilaparvata Lugens ◽

Cytochrome P450 Monooxygenase ◽

P450 Monooxygenase

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Genomic and transcriptomic insights into the cytochrome P450 monooxygenase gene repertoire in the rice pest brown planthopper, Nilaparvata lugens

Genomics ◽

10.1016/j.ygeno.2015.07.010 ◽

2015 ◽

Vol 106 (5) ◽

pp. 301-309 ◽

Cited By ~ 30

Author(s):

Shu-Hua Lao ◽

Xiao-Hui Huang ◽

Hai-Jian Huang ◽

Cheng-Wen Liu ◽

Chuan-Xi Zhang ◽

...

Keyword(s):

Cytochrome P450 ◽

Brown Planthopper ◽

Nilaparvata Lugens ◽

Cytochrome P450 Monooxygenase ◽

Gene Repertoire ◽

P450 Monooxygenase ◽

Monooxygenase Gene ◽

Rice Pest

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Cytochrome P450 Initiates Degradation ofcis-Dichloroethene by Polaromonas sp. Strain JS666

Applied and Environmental Microbiology ◽

10.1128/aem.03445-12 ◽

2013 ◽

Vol 79 (7) ◽

pp. 2263-2272 ◽

Cited By ~ 19

Author(s):

Shirley F. Nishino ◽

Kwanghee A. Shin ◽

James M. Gossett ◽

Jim C. Spain

Keyword(s):

Cytochrome P450 ◽

Heterologous Gene Expression ◽

Degradation Mechanism ◽

Cytochrome P450 Monooxygenase ◽

Cytochrome P450 Enzymes ◽

Catabolic Pathway ◽

P450 Monooxygenase ◽

Content Type ◽

P450 Gene ◽

Cell Extracts

ABSTRACTPolaromonassp. strain JS666 grows oncis-1,2-dichoroethene (cDCE) as the sole carbon and energy source under aerobic conditions, but the degradation mechanism and the enzymes involved are unknown. In this study, we established the complete pathway forcDCE degradation through heterologous gene expression, inhibition studies, enzyme assays, and analysis of intermediates. Several lines of evidence indicate that a cytochrome P450 monooxygenase catalyzes the initial step ofcDCE degradation. Both the transient accumulation of dichloroacetaldehyde incDCE-degrading cultures and dichloroacetaldehyde dehydrogenase activities in cell extracts of JS666 support a pathway for degradation ofcDCE through dichloroacetaldehyde. The mechanism minimizes the formation ofcDCE epoxide. The molecular phylogeny of the cytochrome P450 gene and the organization of neighboring genes suggest that thecDCE degradation pathway recently evolved in a progenitor capable of degrading 1,2-dichloroethane either by the recruitment of the cytochrome P450 monooxygenase gene from an alkane catabolic pathway or by selection for variants of the P450 in a preexisting 1,2-dichloroethane catabolic pathway. The results presented here add yet another role to the broad array of productive reactions catalyzed by cytochrome P450 enzymes.

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Molecular evolution of a cytochrome P450 for the synthesis of potential antidepressant (2R,6R)-hydroxynorketamine

Chemical Communications ◽

10.1039/d0cc06729f ◽

2021 ◽

Author(s):

Ansgar Bokel ◽

Michael C. Hutter ◽

Vlada B. Urlacher

Keyword(s):

Cytochrome P450 ◽

Molecular Evolution ◽

Cytochrome P450 Monooxygenase ◽

P450 Monooxygenase ◽

Effective Synthesis

Engineered cytochrome P450 monooxygenase CYP154E1 enables the effective synthesis of the potential antidepressant (2R,6R)-hydroxynorketamine via N-demethylation and regio- and stereoselective hydroxylation of (R)-ketamine.

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Cytochrome P450 Monooxygenase-Mediated Metabolic Utilization of Benzo[a]Pyrene by Aspergillus Species

mBio ◽

10.1128/mbio.00558-19 ◽

2019 ◽

Vol 10 (3) ◽

Cited By ~ 3

Author(s):

Erin M. Ostrem Loss ◽

Mi-Kyung Lee ◽

Ming-Yueh Wu ◽

Julia Martien ◽

Wanping Chen ◽

...

Keyword(s):

Cytochrome P450 ◽

Environmental Pollutants ◽

Energy Generation ◽

Fungal Species ◽

Metabolic Changes ◽

Cytochrome P450 Monooxygenase ◽

Cellular Growth ◽

P450 Monooxygenase ◽

Content Type ◽

Fundamental Knowledge

ABSTRACT Soil-dwelling fungal species possess the versatile metabolic capability to degrade complex organic compounds that are toxic to humans, yet the mechanisms they employ remain largely unknown. Benzo[a]pyrene (BaP) is a pervasive carcinogenic contaminant, posing a significant concern for human health. Here, we report that several Aspergillus species are capable of degrading BaP. Exposing Aspergillus nidulans cells to BaP results in transcriptomic and metabolic changes associated with cellular growth and energy generation, implying that the fungus utilizes BaP as a growth substrate. Importantly, we identify and characterize the conserved bapA gene encoding a cytochrome P450 monooxygenase that is necessary for the metabolic utilization of BaP in Aspergillus. We further demonstrate that the fungal NF-κB-type velvet regulators VeA and VelB are required for proper expression of bapA in response to nutrient limitation and BaP degradation in A. nidulans. Our study illuminates fundamental knowledge of fungal BaP metabolism and provides novel insights into enhancing bioremediation potential. IMPORTANCE We are increasingly exposed to environmental pollutants, including the carcinogen benzo[a]pyrene (BaP), which has prompted extensive research into human metabolism of toxicants. However, little is known about metabolic mechanisms employed by fungi that are able to use some toxic pollutants as the substrates for growth, leaving innocuous by-products. This study systemically demonstrates that a common soil-dwelling fungus is able to use benzo[a]pyrene as food, which results in expression and metabolic changes associated with growth and energy generation. Importantly, this study reveals key components of the metabolic utilization of BaP, notably a cytochrome P450 monooxygenase and the fungal NF-κB-type transcriptional regulators. Our study advances fundamental knowledge of fungal BaP metabolism and provides novel insight into designing and implementing enhanced bioremediation strategies.

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