scholarly journals Isolation of a gene cluster from Armillaria gallica for the synthesis of armillyl orsellinate–type sesquiterpenoids

2020 ◽  
Vol 105 (1) ◽  
pp. 211-224
Author(s):  
Benedikt Engels ◽  
Uwe Heinig ◽  
Christopher McElroy ◽  
Reinhard Meusinger ◽  
Torsten Grothe ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Gene Clusters ◽  
Bioactive Metabolites ◽  
Cytochrome P450 Monooxygenases ◽  
Cytotoxic Activities ◽  
Fungal Enzymes ◽  
Parasitic Fungi ◽  
P450 Monooxygenases ◽  
Sesquiterpene Cyclase ◽  
Armillaria Gallica

Abstract Melleolides and armillyl orsellinates are protoilludene-type aryl esters that are synthesized exclusively by parasitic fungi of the globally distributed genus Armillaria (Agaricomycetes, Physalacriaceae). Several of these compounds show potent antimicrobial and cytotoxic activities, making them promising leads for the development of new antibiotics or drugs for the treatment of cancer. We recently cloned and characterized the Armillaria gallica gene Pro1 encoding protoilludene synthase, a sesquiterpene cyclase catalyzing the pathway-committing step to all protoilludene-type aryl esters. Fungal enzymes representing secondary metabolic pathways are sometimes encoded by gene clusters, so we hypothesized that the missing steps in the pathway to melleolides and armillyl orsellinates might be identified by cloning the genes surrounding Pro1. Here we report the isolation of an A. gallica gene cluster encoding protoilludene synthase and four cytochrome P450 monooxygenases. Heterologous expression and functional analysis resulted in the identification of protoilludene-8α-hydroxylase, which catalyzes the first committed step in the armillyl orsellinate pathway. This confirms that ∆-6-protoilludene is a precursor for the synthesis of both melleolides and armillyl orsellinates, but the two pathways already branch at the level of the first oxygenation step. Our results provide insight into the synthesis of these valuable natural products and pave the way for their production by metabolic engineering. Key points • Protoilludene-type aryl esters are bioactive metabolites produced by Armillaria spp. • The pathway-committing step to these compounds is catalyzed by protoilludene synthase. • We characterized CYP-type enzymes in the cluster and identified novel intermediates.

Cytochrome P450 Monooxygenases and Interactions of Psychotropic Drugs: A Five-Year Update

1995 ◽  
Vol 25 (3) ◽  
pp. 277-290 ◽  
Author(s):  
Winston W. Shen
Keyword(s):  
Cytochrome P450 ◽  
Psychotropic Drugs ◽  
Selective Serotonin Reuptake Inhibitors ◽  
Gene Clusters ◽  
Serum Levels ◽  
Cytochrome P450s ◽  
Cytochrome P450 Monooxygenases ◽  
P450 Monooxygenases ◽  
Human Cytochrome ◽  
Enzymatic Induction

Objective: This article is a five-year update on a previous review article ( International Journal of Psychiatry in Medicine, 21:47–56, 1991) on cytochrome P450 monooxygenases and interactions of psychotropic drugs. Method: In the literature review, the recent committee work on nomenclature of the P450 superfamily are highlighted. Then, the author reviewed gene clusters of three human cytochrome P450s— CYP1A2, CYP2D6, and CYP3A4 with the focus on the changes of serum levels of the coadministered psychotropic drugs in the context of enzymatic induction and inhibition of these three hepatic enzymes. Results: As indicated in one table, the author stratified probes, inducers, inhibitors, chemical reactions, and substrates under these three gene clusters. As shown in another simple table, the author compared the hepatic enzymatic inhibitions of four selective serotonin reuptake inhibitors and pointed out the inhibition potentials of fluvoxamine at CYP1A2, fluoxetine and paroxetine at CYP2D6, and fluoxetine and fluvoxamine at CYP3A4 if these two SSRIs have higher serum concentrations. Conclusion: The author suggests that with these systematic approaches, this rapidly adding knowledge can help psychiatrists better understand psychotropic drug interactions and maximize the benefits of patients' psychopharmacotherapy.

scholarly journals Comparative Phylogenomic Analysis of Cytochrome P450 Monooxygenases From Fusarium Species

2021 ◽  
Author(s):  
Wadzani Palnam Dauda ◽  
Elkanah Glen ◽  
Peter Abraham ◽  
Charles Oluwaseun Adetunji ◽  
Daji Morumda ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Polyketide Synthase ◽  
Fusarium Species ◽  
Pathogenic Fungi ◽  
Plant Diseases ◽  
Biological Research ◽  
Cytochrome P450 Monooxygenases ◽  
Phylogenomic Analysis ◽  
P450 Monooxygenases

Abstract Cytochrome P450s (P450s) are a unique multifamily class of enzymes that possess the capability to exhibit catalytic versatility in several biochemical reactions which entails metabolite biosynthesis, primary and secondary metabolism. Fusarium spp. is an important microorganism with many members known to produce secondary metabolites that cause plant diseases and mycotoxicoses in animals and humans. In this present study, from the initially screened 4,579 proteins, we elucidated the nature of abundance, evolutionary relationships, classification and cellular location of 320 cytochrome P450 from 17 phytopathogenic members of Fusarium species. The total CYPs protein sequences were phylogenetically grouped into seventeen (17) clades. Eighty-six (86) CYPs families and forty-eight (48) clans were identified. Twenty-seven (27) families were each found in only one species. The CYPs were found to be majorly localized in the endoplasmic reticulum. The non-ribosomal peptide synthetase-like (NRPS-like) gene cluster was the predominant secondary metabolic-related gene cluster across all the seventeen selected Fusarium species except in F. cucurbiticola and F. solani, where PolyKetide Synthase (PKS) was the most prevalent. The presence of numerous families and clans as observed in in this study shows the expansions of the CYPs families across Fusarium species, this CYPs family and clan expansion is often associated with the evolvement of several fungal traits that include their pathogenicity adaptation to survive on an extensive range of toxic substrates. Identification of P450 proteins in these pathogenic fungi provides fundamental information for further basic and applied biological research into the physiological and toxigenic roles of P450s in Fusarium species.

scholarly journals Comparative Analyses of Cytochrome P450s and Those Associated with Secondary Metabolism in Bacillus Species

2018 ◽  
Vol 19 (11) ◽  
pp. 3623 ◽  
Author(s):  
Bongumusa Mthethwa ◽  
Wanping Chen ◽  
Mathula Ngwenya ◽  
Abidemi Kappo ◽  
Puleng Syed ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Secondary Metabolite ◽  
In Silico Analysis ◽  
Gene Clusters ◽  
Bacillus Species ◽  
Cytochrome P450 Monooxygenases ◽  
Enzymatic Reactions ◽  
P450 Monooxygenases ◽  
Stereo Selectivity

Cytochrome P450 monooxygenases (CYPs/P450s) are among the most catalytically-diverse enzymes, capable of performing enzymatic reactions with chemo-, regio-, and stereo-selectivity. Our understanding of P450s’ role in secondary metabolite biosynthesis is becoming broader. Among bacteria, Bacillus species are known to produce secondary metabolites, and recent studies have revealed the presence of secondary metabolite biosynthetic gene clusters (BGCs) in these species. However, a comprehensive comparative analysis of P450s and P450s involved in the synthesis of secondary metabolites in Bacillus species has not been reported. This study intends to address these two research gaps. In silico analysis of P450s in 128 Bacillus species revealed the presence of 507 P450s that can be grouped into 13 P450 families and 28 subfamilies. No P450 family was found to be conserved in Bacillus species. Bacillus species were found to have lower numbers of P450s, P450 families and subfamilies, and a lower P450 diversity percentage compared to mycobacterial species. This study revealed that a large number of P450s (112 P450s) are part of different secondary metabolite BGCs, and also identified an association between a specific P450 family and secondary metabolite BGCs in Bacillus species. This study opened new vistas for further characterization of secondary metabolite BGCs, especially P450s in Bacillus species.

scholarly journals Comprehensive Analyses of Cytochrome P450 Monooxygenases and Secondary Metabolite Biosynthetic Gene Clusters in Cyanobacteria

2020 ◽  
Vol 21 (2) ◽  
pp. 656 ◽  
Author(s):  
Makhosazana Jabulile Khumalo ◽  
Nomfundo Nzuza ◽  
Tiara Padayachee ◽  
Wanping Chen ◽  
Jae-Hyuk Yu ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Secondary Metabolites ◽  
Secondary Metabolite ◽  
Gene Clusters ◽  
Cytochrome P450 Monooxygenases ◽  
Biosynthetic Gene ◽  
Mycobacterial Species ◽  
Biosynthetic Gene Clusters ◽  
P450 Monooxygenases ◽  
Cyanobacterial Species

The prokaryotic phylum Cyanobacteria are some of the oldest known photosynthetic organisms responsible for the oxygenation of the earth. Cyanobacterial species have been recognised as a prosperous source of bioactive secondary metabolites with antibacterial, antiviral, antifungal and/or anticancer activities. Cytochrome P450 monooxygenases (CYPs/P450s) contribute to the production and diversity of various secondary metabolites. To better understand the metabolic potential of cyanobacterial species, we have carried out comprehensive analyses of P450s, predicted secondary metabolite biosynthetic gene clusters (BGCs), and P450s located in secondary metabolite BGCs. Analysis of the genomes of 114 cyanobacterial species identified 341 P450s in 88 species, belonging to 36 families and 79 subfamilies. In total, 770 secondary metabolite BGCs were found in 103 cyanobacterial species. Only 8% of P450s were found to be part of BGCs. Comparative analyses with other bacteria Bacillus, Streptomyces and mycobacterial species have revealed a lower number of P450s and BGCs and a percentage of P450s forming part of BGCs in cyanobacterial species. A mathematical formula presented in this study revealed that cyanobacterial species have the highest gene-cluster diversity percentage compared to Bacillus and mycobacterial species, indicating that these diverse gene clusters are destined to produce different types of secondary metabolites. The study provides fundamental knowledge of P450s and those associated with secondary metabolism in cyanobacterial species, which may illuminate their value for the pharmaceutical and cosmetics industries.

scholarly journals Ancient Bacterial Class Alphaproteobacteria Cytochrome P450 Monooxygenases Can Be Found in Other Bacterial Species

2021 ◽  
Vol 22 (11) ◽  
pp. 5542
Author(s):  
Nomfundo Nzuza ◽  
Tiara Padayachee ◽  
Puleng Rosinah Syed ◽  
Justyna Dorota Kryś ◽  
Wanping Chen ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Bacterial Species ◽  
Gene Clusters ◽  
Side Chain ◽  
Cytochrome P450 Monooxygenases ◽  
Biosynthetic Gene Clusters ◽  
Genome Data ◽  
P450 Monooxygenases ◽  
Large Numbers ◽  
Cyanobacterial Species

Cytochrome P450 monooxygenases (CYPs/P450s), heme-thiolate proteins, are well-known players in the generation of chemicals valuable to humans and as a drug target against pathogens. Understanding the evolution of P450s in a bacterial population is gaining momentum. In this study, we report comprehensive analysis of P450s in the ancient group of the bacterial class Alphaproteobacteria. Genome data mining and annotation of P450s in 599 alphaproteobacterial species belonging to 164 genera revealed the presence of P450s in only 241 species belonging to 82 genera that are grouped into 143 P450 families and 214 P450 subfamilies, including 77 new P450 families. Alphaproteobacterial species have the highest average number of P450s compared to Firmicutes species and cyanobacterial species. The lowest percentage of alphaproteobacterial species P450s (2.4%) was found to be part of secondary metabolite biosynthetic gene clusters (BGCs), compared other bacterial species, indicating that during evolution large numbers of P450s became part of BGCs in other bacterial species. Our study identified that some of the P450 families found in alphaproteobacterial species were passed to other bacterial species. This is the first study to report on the identification of CYP125 P450, cholesterol and cholest-4-en-3-one hydroxylase in alphaproteobacterial species (Phenylobacterium zucineum) and to predict cholesterol side-chain oxidation capability (based on homolog proteins) by P. zucineum.

scholarly journals More P450s Are Involved in Secondary Metabolite Biosynthesis in Streptomyces Compared to Bacillus, Cyanobacteria, and Mycobacterium

2020 ◽  
Vol 21 (13) ◽  
pp. 4814
Author(s):  
Fanele Cabangile Mnguni ◽  
Tiara Padayachee ◽  
Wanping Chen ◽  
Dominik Gront ◽  
Jae-Hyuk Yu ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Secondary Metabolite ◽  
Gene Clusters ◽  
Bacillus Species ◽  
Cytochrome P450 Monooxygenases ◽  
Streptomyces Species ◽  
P450 Monooxygenases ◽  
Cyanobacterial Species ◽  
Metabolite Synthesis

Unraveling the role of cytochrome P450 monooxygenases (CYPs/P450s), heme-thiolate proteins present in living and non-living entities, in secondary metabolite synthesis is gaining momentum. In this direction, in this study, we analyzed the genomes of 203 Streptomyces species for P450s and unraveled their association with secondary metabolism. Our analyses revealed the presence of 5460 P450s, grouped into 253 families and 698 subfamilies. The CYP107 family was found to be conserved and highly populated in Streptomyces and Bacillus species, indicating its key role in the synthesis of secondary metabolites. Streptomyces species had a higher number of P450s than Bacillus and cyanobacterial species. The average number of secondary metabolite biosynthetic gene clusters (BGCs) and the number of P450s located in BGCs were higher in Streptomyces species than in Bacillus, mycobacterial, and cyanobacterial species, corroborating the superior capacity of Streptomyces species for generating diverse secondary metabolites. Functional analysis via data mining confirmed that many Streptomyces P450s are involved in the biosynthesis of secondary metabolites. This study was the first of its kind to conduct a comparative analysis of P450s in such a large number (203) of Streptomyces species, revealing the P450s’ association with secondary metabolite synthesis in Streptomyces species. Future studies should include the selection of Streptomyces species with a higher number of P450s and BGCs and explore the biotechnological value of secondary metabolites they produce.

scholarly journals A Single Biosynthetic Gene Cluster Is Responsible for the Production of Bagremycin Antibiotics and Ferroverdin Iron Chelators

mBio ◽  
2019 ◽  
Vol 10 (4) ◽  
Author(s):  
Loïc Martinet ◽  
Aymeric Naômé ◽  
Benoit Deflandre ◽  
Marta Maciejewska ◽  
Déborah Tellatin ◽  
...  
Keyword(s):  
Natural Product ◽  
Gene Cluster ◽  
Biosynthetic Pathway ◽  
Genome Mining ◽  
Gene Clusters ◽  
Bioactive Molecules ◽  
Bioactive Metabolites ◽  
Biosynthetic Gene ◽  
Single Family ◽  
Biosynthetic Gene Clusters

ABSTRACT Biosynthetic gene clusters (BGCs) are organized groups of genes involved in the production of specialized metabolites. Typically, one BGC is responsible for the production of one or several similar compounds with bioactivities that usually only vary in terms of strength and/or specificity. Here we show that the previously described ferroverdins and bagremycins, which are families of metabolites with different bioactivities, are produced from the same BGC, whereby the fate of the biosynthetic pathway depends on iron availability. Under conditions of iron depletion, the monomeric bagremycins are formed, representing amino-aromatic antibiotics resulting from the condensation of 3-amino-4-hydroxybenzoic acid with p-vinylphenol. Conversely, when iron is abundantly available, the biosynthetic pathway additionally produces a molecule based on p-vinylphenyl-3-nitroso-4-hydroxybenzoate, which complexes iron to form the trimeric ferroverdins that have anticholesterol activity. Thus, our work shows a unique exception to the concept that BGCs should only produce a single family of molecules with one type of bioactivity and that in fact different bioactive molecules may be produced depending on the environmental conditions. IMPORTANCE Access to whole-genome sequences has exposed the general incidence of the so-called cryptic biosynthetic gene clusters (BGCs), thereby renewing their interest for natural product discovery. As a consequence, genome mining is the often first approach implemented to assess the potential of a microorganism for producing novel bioactive metabolites. By revealing a new level of complexity of natural product biosynthesis, we further illustrate the difficulty of estimation of the panel of molecules associated with a BGC based on genomic information alone. Indeed, we found that the same gene cluster is responsible for the production of compounds which differ in terms of structure and bioactivity. The production of these different compounds responds to different environmental triggers, which suggests that multiplication of culture conditions is essential for revealing the entire panel of molecules made by a single BGC.

scholarly journals Omega-3 fatty acid-derived mediators that control inflammation and tissue homeostasis

2019 ◽  
Vol 31 (9) ◽  
pp. 559-567 ◽  
Author(s):  
Tomoaki Ishihara ◽  
Mio Yoshida ◽  
Makoto Arita
Keyword(s):  
Fatty Acid ◽  
Docosapentaenoic Acid ◽  
Tissue Homeostasis ◽  
Bioactive Metabolites ◽  
Cytochrome P450 Monooxygenases ◽  
Biological Functions ◽  
Omega 3 ◽  
Beneficial Effects ◽  
P450 Monooxygenases ◽  
Wide Range

AbstractOmega-3 polyunsaturated fatty acids (PUFAs), including eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid, display a wide range of beneficial effects in humans and animals. Many of the biological functions of PUFAs are mediated via bioactive metabolites produced by fatty acid oxygenases such as cyclooxygenases, lipoxygenases and cytochrome P450 monooxygenases. Liquid chromatography–tandem mass spectrometry-based mediator lipidomics revealed a series of novel bioactive lipid mediators derived from omega-3 PUFAs. Here, we describe recent advances on omega-3 PUFA-derived mediators, mainly focusing on their enzymatic oxygenation pathway, and their biological functions in controlling inflammation and tissue homeostasis.

scholarly journals A biaryl-linked tripeptide from Planomonospora leads to widespread class of minimal RiPP gene clusters

2020 ◽  
Author(s):  
Mitja M. Zdouc ◽  
Mohammad M. Alanjary ◽  
Guadalupe S. Zarazúa ◽  
Sonia I. Maffioli ◽  
Max Crüsemann ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Structural Diversity ◽  
Gene Clusters ◽  
Bioinformatic Analysis ◽  
Cytochrome P450 Monooxygenases ◽  
Bacterial Genomes ◽  
P450 Monooxygenase ◽  
Linked Gene ◽  
P450 Monooxygenases ◽  
Microbial Natural Products

AbstractMicrobial natural products impress by their bioactivity, structural diversity and ingenious biosynthesis. While screening the rare actinobacterial genus Planomonospora, cyclopeptides 1A and 1B were discovered, featuring an unusual Tyr-His biaryl-bridging across a tripeptide scaffold, with the sequences N-acetyl-Tyr-Tyr-His (1A) and N-acetyl-Tyr-Phe-His (1B). Genome analysis of the 1A producing strain pointed to-wards a ribosomal synthesis of 1A, from a pentapeptide precursor encoded by the tiny 18-nucleotide gene bycA, to our knowledge the smallest gene ever reported. Further, biaryl instalment is performed by the closely linked gene bycB, encoding a cytochrome P450 monooxygenase. Biosynthesis of 1A was confirmed by heterologous production in Streptomyces, yielding the mature product. Bioinformatic analysis of related cytochrome P450 monooxygenases indicated that they constitute a widespread family of pathways, associated to 5-aa coding sequences in approximately 200 (actino)bacterial genomes, all with potential for a biaryl linkage between amino acids 1 and 3. We propose the name biarylicins for this newly discovered family of RiPPs.

scholarly journals Isolation and Characterization of the Gibberellin Biosynthetic Gene Cluster in Sphaceloma manihoticola

2008 ◽  
Vol 74 (17) ◽  
pp. 5325-5339 ◽  
Author(s):  
Christiane Bömke ◽  
Maria Cecilia Rojas ◽  
Fan Gong ◽  
Peter Hedden ◽  
Bettina Tudzynski
Keyword(s):  
Gene Cluster ◽  
Gene Clusters ◽  
Biosynthetic Gene Cluster ◽  
Gene Replacement ◽  
Gibberella Fujikuroi ◽  
Cytochrome P450 Monooxygenases ◽  
Fusarium Fujikuroi ◽  
Biosynthetic Gene ◽  
Isolation And Characterization ◽  
Gene Functions

ABSTRACT Gibberellins (GAs) are tetracyclic diterpenoid phytohormones that were first identified as secondary metabolites of the fungus Fusarium fujikuroi (teleomorph, Gibberella fujikuroi). GAs were also found in the cassava pathogen Sphaceloma manihoticola, but the spectrum of GAs differed from that in F. fujikuroi. In contrast to F. fujikuroi, the GA biosynthetic pathway has not been studied in detail in S. manihoticola, and none of the GA biosynthetic genes have been cloned from the species. Here, we present the identification of the GA biosynthetic gene cluster from S. manihoticola consisting of five genes encoding a bifunctional ent-copalyl/ent-kaurene synthase (CPS/KS), a pathway-specific geranylgeranyl diphosphate synthase (GGS2), and three cytochrome P450 monooxygenases. The functions of all of the genes were analyzed either by a gene replacement approach or by complementing the corresponding F. fujikuroi mutants. The cluster organization and gene functions are similar to those in F. fujikuroi. However, the two border genes in the Fusarium cluster encoding the GA4 desaturase (DES) and the 13-hydroxylase (P450-3) are absent in the S. manihoticola GA gene cluster, consistent with the spectrum of GAs produced by this fungus. The close similarity between the two GA gene clusters, the identical gene functions, and the conserved intron positions suggest a common evolutionary origin despite the distant relatedness of the two fungi.

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