Tirandamycin biosynthesis is mediated by co-dependent oxidative enzymes

Carlson, Jacob C.; Li, Shengying; Gunatilleke, Shamila S.; Anzai, Yojiro; Burr, Douglas A.; Podust, L.M.; Sherman, David H.

doi:10.1038/nchem.1087

Cited by 86 publications

(141 citation statements)

References 31 publications

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“…This is similar to what we have observed with CreJEF, CreC, and CreG. Likewise, in the tirandamycin biosynthetic pathway, TamI P450 enzyme employs the FAD-containing TamL oxidase to overcome the conversion from an alcohol to a keto group (63). These examples suggest that the successive oxidations are likely a heavy burden for P450 enzymes to accomplish on their own; therefore, assistance is required from other types of oxidases.…”

Section: Discussionsupporting

confidence: 64%

Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum

et al. 2016

Journal of Biological Chemistry

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4-Cresol is not only a significant synthetic intermediate for production of many aromatic chemicals, but also a priority environmental pollutant because of its toxicity to higher organisms. In our previous studies, a gene cluster implicated to be involved in 4-cresol catabolism, creCDEFGHIR, was identified in Corynebacterium glutamicum and partially characterized in vivo. In this work, we report on the discovery of a novel 4-cresol biodegradation pathway that employs phosphorylated intermediates. This unique pathway initiates with the phosphorylation of the hydroxyl group of 4-cresol, which is catalyzed by a novel 4-methylbenzyl phosphate synthase, CreHI. Next, a unique class I P450 system, CreJEF, specifically recognizes phosphorylated intermediates and successively oxidizes the aromatic methyl group into carboxylic acid functionality via alcohol and aldehyde intermediates. Moreover, CreD (phosphohydrolase), CreC (alcohol dehydrogenase), and CreG (aldehyde dehydrogenase) were also found to be required for efficient oxidative transformations in this pathway. Steady-state kinetic parameters (K m and k cat ) for each catabolic step were determined, and these results suggest that kinetic controls serve a key role in directing the metabolic flux to the most energy effective route.

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Section: Discussionsupporting

confidence: 64%

Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum

et al. 2016

Journal of Biological Chemistry

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“…This result has important implications for metabolic engineering of crop plants for enhanced disease resistance and also for the generation of antimicrobial triterpenes for other applications. Multifunctional P450 enzymes from other P450 families that catalyze both hydroxylation and epoxidation reactions have been described recently from bacteria (47)(48)(49)(50)(51) and fungi (52,53). Examples of both possible reaction orders (hydroxylation followed by epoxidation and epoxidation followed by hydroxylation) have been reported (Fig.…”

Section: Resultsmentioning

confidence: 99%

Biochemical analysis of a multifunctional cytochrome P450 (CYP51) enzyme required for synthesis of antimicrobial triterpenes in plants

Geisler

Hughes

Sainsbury

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

148

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Members of the cytochromes P450 superfamily (P450s) catalyze a huge variety of oxidation reactions in microbes and higher organisms. Most P450 families are highly divergent, but in contrast the cytochrome P450 14α-sterol demethylase (CYP51) family is one of the most ancient and conserved, catalyzing sterol 14α-demethylase reactions required for essential sterol synthesis across the fungal, animal, and plant kingdoms. Oats (Avena spp.) produce antimicrobial compounds, avenacins, that provide protection against disease. Avenacins are synthesized from the simple triterpene, β-amyrin. Previously we identified a gene encoding a member of the CYP51 family of cytochromes P450, AsCyp51H10 (also known as Saponin-deficient 2, Sad2), that is required for avenacin synthesis in a forward screen for avenacin-deficient oat mutants. sad2 mutants accumulate β-amyrin, suggesting that they are blocked early in the pathway. Here, using a transient plant expression system, we show that AsCYP51H10 is a multifunctional P450 capable of modifying both the C and D rings of the pentacyclic triterpene scaffold to give 12,13β-epoxy-3β,16β-dihydroxy-oleanane (12,13β-epoxy-16β-hydroxy-β-amyrin). Molecular modeling and docking experiments indicate that C16 hydroxylation is likely to precede C12,13 epoxidation. Our computational modeling, in combination with analysis of a suite of sad2 mutants, provides insights into the unusual catalytic behavior of AsCYP51H10 and its active site mutants. Fungal bioassays show that the C12,13 epoxy group is an important determinant of antifungal activity. Accordingly, the oat AsCYP51H10 enzyme has been recruited from primary metabolism and has acquired a different function compared to other characterized members of the plant CYP51 family-as a multifunctional stereo-and regio-specific hydroxylase in plant specialized metabolism.

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“…[166][167][168] Other complex examples of multi-step P450 reactions (combining hydroxylation and epoxidation) have been reported in the biosynthesis of the macrolides FD-891, 169 mycinamycin (MycG) 170 and tirandamycin (TamI). 171 In the case of MycG, the two-step transformation preferentially begins with hydroxylation at C14, with subsequent epoxidation of the C12-C13 double bond (Fig. 16B).…”

Section: Complex Transformations Mediated By P450s In Pks Biosynthesismentioning

confidence: 99%

Unrivalled diversity: the many roles and reactions of bacterial cytochromes P450 in secondary metabolism

et al. 2018

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The cytochromes P450 (P450s) are a superfamily of heme-containing monooxygenases that perform diverse catalytic roles in many species, including bacteria. The P450 superfamily is widely known for the hydroxylation of unactivated C-H bonds, but the diversity of reactions that P450s can perform vastly exceeds this undoubtedly impressive chemical transformation. Within bacteria, P450s play important roles in many biosynthetic and biodegradative processes that span a wide range of secondary metabolite pathways and present diverse chemical transformations. In this review, we aim to provide an overview of the range of chemical transformations that P450 enzymes can catalyse within bacterial secondary metabolism, with the intention to provide an important resource to aid in understanding of the potential roles of P450 enzymes within newly identified bacterial biosynthetic pathways. 1.Introduction to P450s 2.Chemistry of P450 enzymes 3.Bacterial biosynthesis pathways involving P450s 3.1Terpene metabolism 3.1. Battersby (1925Battersby ( -2018 to the molecular understanding of biosynthesis over the course of their careers.

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Tirandamycin biosynthesis is mediated by co-dependent oxidative enzymes

Cited by 86 publications

References 31 publications

Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum

Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum

Biochemical analysis of a multifunctional cytochrome P450 (CYP51) enzyme required for synthesis of antimicrobial triterpenes in plants

Unrivalled diversity: the many roles and reactions of bacterial cytochromes P450 in secondary metabolism

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