Substituent Effects on the Phenol Coupling Reaction Catalyzed by the Vancomycin Biosynthetic P450 Enzyme OxyB

Schmartz, Patrick C.; Wölfel, Katharina; Zerbe, Katja; Gad, Emad M.; Tamany, El Sayed El; Ibrahim, H. K.; Abou‐Hadeed, K.

doi:10.1002/anie.201204458

Cited by 33 publications

(39 citation statements)

References 30 publications

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“…9 It has been shown that OxyB exhibits a marked preference for the unchlorinated substrate, but the order of chlorination with respect to the transformations catalyzed by OxyA and OxyC still awaits clarification. 19 We therefore hypothesized that the correct substrate was not available in our reaction mixtures above, and that OxyA may only recognize the singly cross-linked bis-chlorinated 7mer peptide. To test this idea, we synthesized the 7mer substrate, now containing 3-Cl-L-Tyr ( 2 , 5 ) and 3-Cl-D-Tyr ( 3 , 5 ) at residues 2 and 6, respectively (Figure 2A, Figures S6–S10), and subsequently performed assays initially only with OxyB.…”

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confidence: 99%

“…8 OxyB can also act on the 6mer product bound to the penultimate or ultimate PCP domain, but it shows a higher k cat / K m toward the 7mer substrate. 11,19 Deletion of oxyB in A. mediterranei results in the accumulation of a linear, unchlorinated, β -OH-Tyr-containing substrate, indicating that β -hydroxylation occurs before OxyB-mediated cross-link formation, while β -OH-Tyr-chlorination occurs after. 9 In accordance, in vitro assays of OxyB show a strict preference for substrates lacking Cl at the C-ring and tolerance toward substrates chlorinated on the E ring, which is not involved in OxyB-catalyzed cross-linking.…”

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In Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis

et al. 2017

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Studies on the biosynthesis of glycopeptide antibiotics have provided many insights into the strategies that Nature employs to build architecturally strained molecules. A key structural feature of vancomycin, the founding member of this class, is a set of three aromatic cross-links that are introduced via yet unknown mechanisms. Previous reports have identified three cytochrome P450 enzymes involved in this process and demonstrated enzymatic activity for OxyB, which installs the first aromatic cross-link. However, the activities of the remaining two P450 enzymes have not been recapitulated. Herein, we show that OxyA generates the second bis-aryl ether bond in vancomycin and that it exhibits strict substrate specificity toward the chlorinated, OxyB-cross-linked product. No OxyA product is detected with the unchlorinated substrate. Together with previous results, these data suggest that chlorination occurs after OxyB- but before OxyA-catalyzed cross-link formation. Our results have important implications for the chemo-enzymatic synthesis of vancomycin and its analogs.

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In Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis

et al. 2017

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“…PCP-bound) peptides, which was conrmed for the OxyB enzyme from vancomycin biosynthesis by the work of Robinson and co-workers. [250][251][252] More recent work has shown that the process of P450 recruitment to the NRPS-bound peptide is more complicated than is the case for PCP-bound amino acids, with GPA biosynthesis relying on the X-domain, a conserved (albeit modied) condensation domain found between the PCP 253 and TE-domains in the nal NRPS module. 243,[254][255][256][257] The X-domain has an interaction interface that specically binds to the Oxy enzymes, with these continually binding to and releasing from the X-domain to ensure that complete crosslinking of the peptide occurs.…”

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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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“…Vancomycin and related antibiotics contain aryl C-C and C-O crosslinks catalyzed by P450-mediated 1-electron oxidations (Figure 2, green). Recent work on the biosynthesis of these antibiotics includes the solution of two crystal structures of P450s involved in aryl coupling reactions [25,26], as well as a study that examines the timing of P450-catalyzed crosslinking during vancomycin biosynthesis [27]. …”

Section: Intermediates In the P450 Catalytic Cycle Drive Diverse Natumentioning

confidence: 99%

“…In pink are highlighted the iron-peroxo and iron-hydroperoxy intermediates and C-C bond scission [30] and sulfoxidation [14] reactions. Highlighted in green are compound I and desaturation [23•], ring closure [17], sequential oxidation [22], and aryl coupling reactions [27]. …”

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Expanding P450 catalytic reaction space through evolution and engineering

McIntosh

Farwell

Arnold

2014

Current Opinion in Chemical Biology

128

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Advances in protein and metabolic engineering have led to wider use of enzymes to synthesize important molecules. However, many desirable transformations are not catalyzed by any known enzyme, driving interest in understanding how new enzymes can be created. The cytochrome P450 enzyme family, whose members participate in xenobiotic metabolism and natural products biosynthesis, catalyzes an impressive range of difficult chemical reactions that continues to grow as new enzymes are characterized. Recent work has revealed that P450-derived enzymes can also catalyze useful reactions previously accessible only to synthetic chemistry. The evolution and engineering of these enzymes provides an excellent case study for how to genetically encode new chemistry and expand biology’s reaction space.

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Substituent Effects on the Phenol Coupling Reaction Catalyzed by the Vancomycin Biosynthetic P450 Enzyme OxyB

Cited by 33 publications

References 30 publications

In Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis

In Vitro Reconstitution of OxyA Enzymatic Activity Clarifies Late Steps in Vancomycin Biosynthesis

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

Expanding P450 catalytic reaction space through evolution and engineering

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