The Cytochrome P450 Complement (CYPome) of Streptomyces coelicolor A3(2)

Lamb, David C.; Skaug, Tove; Song, Honglin; Jackson, C.J.; Podust, L.M.; Waterman, Michael R.; Kell, Douglas B.; Kelly, Diane; Kelly, Steven L.

doi:10.1074/jbc.m111109200

Cited by 124 publications

(112 citation statements)

References 25 publications

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“…Sequence Homology with Other Monooxygenases-CYP154C1 (Q9L142; accession numbers are according to SWISS-PROT/ TrEMBL Protein Knowledgebase, tw.expasy.org/sprot/) is one of the 18 cytochromes P450 revealed in the S. coelicolor A3 (2) genome (10), and one of the two assigned to the CYP154 family (11). The closest homolog of CYP154C1 identified in the data base using program BLAST (45) is a cytochrome P450 CYP154B1 from Streptomyces fradiae with 44% sequence identity (Q9XCC6) localized within the tylosin biosynthetic gene cluster (13).…”

Section: Resultsmentioning

confidence: 99%

“…CYP154C1 Purification-The DNA sequence corresponding to CYP154C1 with four histidine codons inserted at the 3Ј end was generated by PCR and cloned into the Escherichia coli expression vector pET17b as described elsewhere (11). The protein was expressed in HMS174(DE3) cells (Novagen) and purified to homogeneity by nickelnitrilotriacetic acid (Qiagen) and Q-Sepharose (Amersham Biosciences) chromatography.…”

Section: Methodsmentioning

confidence: 99%

“…Although 18 CYP open reading frames are found dispersed throughout the S. coelicolor A3(2) chromosome (10,11), the functions of the corresponding gene products and their role in various metabolic functions remain largely undefined.…”

mentioning

confidence: 99%

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The 1.92-Å Structure of Streptomyces coelicolor A3(2) CYP154C1

Podust

Kim

Arase

et al. 2003

Journal of Biological Chemistry

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Evolutionary links between cytochrome P450 monooxygenases, a superfamily of extraordinarily divergent heme-thiolate proteins catalyzing a wide array of NADPH/NADH-and O 2 -dependent reactions, are becoming better understood because of availability of an increasing number of fully sequenced genomes. Among other reactions, P450s catalyze the site-specific oxidation of the precursors to macrolide antibiotics in the genus Streptomyces introducing regiochemical diversity into the macrolide ring system, thereby significantly increasing antibiotic activity. Developing effective uses for Streptomyces enzymes in biosynthetic processes and bioremediation requires identification and engineering of additional monooxygenases with activities toward a diverse array of small molecules. To elucidate the molecular basis for substrate specificity of oxidative enzymes toward macrolide antibiotics, the x-ray structure of CYP154C1 from Streptomyces coelicolor A3(2) was determined (Protein Data Bank code 1GWI). Relocation of certain common P450 secondary structure elements, along with a novel structural feature involving an additional ␤-strand transforming the five-stranded ␤-sheet into a six-stranded variant, creates an open cleft-shaped substrate-binding site between the two P450 domains. High sequence similarity to macrolide monooxygenases from other microbial species translates into catalytic activity of CYP154C1 toward both 12-and 14-membered ring macrolactones in vitro.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The 1.92-Å Structure of Streptomyces coelicolor A3(2) CYP154C1

Podust

Kim

Arase

et al. 2003

Journal of Biological Chemistry

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“…The large biodiversity of CYPs is not only found in these wood-rot fungi, but in other saprophytic organisms, e.g. Aspergillus oryzae [68], and in soil actinomycetes such as the 18 CYPs in S. coelicolor A3(2) [19] and approximately 40 in Mycobacterium smegmatis [12,69]. The ability of these organisms to biotransform xenobiotics is well known, including for instance, the ability of aspergilli in industrial steroid hydroxylation.…”

Section: Microbial Cytochromes P450 and The Environmentmentioning

confidence: 99%

Microbial cytochromes P450: biodiversity and biotechnology. Where do cytochromes P450 come from, what do they do and what can they do for us?

Kelly

2013

Phil. Trans. R. Soc. B

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The first eukaryote genome revealed three yeast cytochromes P450 (CYPs), hence the subsequent realization that some microbial fungal genomes encode these proteins in 1 per cent or more of all genes (greater than 100) has been surprising. They are unique biocatalysts undertaking a wide array of stereo-and regio-specific reactions and so hold promise in many applications. Based on ancestral activities that included 14a-demethylation during sterol biosynthesis, it is now seen that CYPs are part of the genes and metabolism of most eukaryotes. In contrast, Archaea and Eubacteria often do not contain CYPs, while those that do are frequently interesting as producers of natural products undertaking their oxidative tailoring. Apart from roles in primary and secondary metabolism, microbial CYPs are actual/potential targets of drugs/agrochemicals and CYP51 in sterol biosynthesis is exhibiting evolution to resistance in the clinic and the field. Other CYP applications include the first industrial biotransformation for corticosteroid production in the 1950s, the diversion into penicillin synthesis in early mutations in fungal strain improvement and bioremediation using bacteria and fungi. The vast untapped resource of orphan CYPs in numerous genomes is being probed and new methods for discovering function and for discovering desired activities are being investigated.

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“…Cytochrome P450 (P450) 4 enzymes catalyze a variety of oxidations and related reactions (2,3). The P450s are found throughout the biological kingdoms, from eubacteria and archebacteria to mammals.…”

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confidence: 99%

Electron Transport Pathway for a Streptomyces Cytochrome P450

Chun

Shimada

Sanchez-Ponce

et al. 2007

Journal of Biological Chemistry

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Streptomyces and other bacterial actinomycete species produce many important natural products, including the majority of known antibiotics, and cytochrome P450 (P450) enzymes catalyze important biosynthetic steps. Relatively few electron transport pathways to P450s have been characterized in bacteria, particularly streptomycete species. One of the 18 P450s in Streptomyces coelicolor A3(2), P450 105D5, was found to bind fatty acids tightly and form hydroxylated products when electrons were delivered from heterologous systems. The six ferredoxin (Fdx) and four flavoprotein Fdx reductase (FDR) proteins coded by genes in S. coelicolor were expressed in Escherichia coli, purified, and used to characterize the electron transfer pathway. Of the many possibilities, the primary pathway was NADH 3 FDR1 3 Fdx4 3 P450 105D5. The genes coding for FDR1, Fdx4, and P450 105D5 are located close together in the S. coelicolor genome. Several fatty acids examined were substrates, including those found in S. coelicolor extracts, and all yielded several products. Mass spectra of the products of lauric acid imply the 8-, 9-, 10-, and 11-hydroxy derivatives. Hydroxylated fatty acids were also detected in vivo in S. coelicolor. Rates of electron transfer between the proteins were measured; all steps were faster than overall hydroxylation and consistent with rates of NADH oxidation. Substrate binding, product release, and oxygen binding were relatively fast in the catalytic cycle; high kinetic deuterium isotope effects for all four lauric acid hydroxylations indicated that the rate of C-H bond breaking is rate-limiting in every case. Thus, an electron transfer pathway to a functional Streptomyces P450 has been established.

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The Cytochrome P450 Complement (CYPome) of Streptomyces coelicolor A3(2)

Cited by 124 publications

References 25 publications

The 1.92-Å Structure of Streptomyces coelicolor A3(2) CYP154C1

The 1.92-Å Structure of Streptomyces coelicolor A3(2) CYP154C1

Microbial cytochromes P450: biodiversity and biotechnology. Where do cytochromes P450 come from, what do they do and what can they do for us?

Electron Transport Pathway for a Streptomyces Cytochrome P450

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