Biosynthetic Timing and Substrate Specificity for the Thiopeptide Thiomuracin

Zhang, Zhengan; Hudson, Graham A.; Mahanta, Nilkamal; Tietz, Jonathan I.; Donk, Wilfred A. van der; Mitchell, Douglas A.

doi:10.1021/jacs.6b08987

Cited by 77 publications

(124 citation statements)

References 18 publications

Supporting

Mentioning

118

Contrasting

Order By: Relevance

“…204,205 A more complex example of amino acid oxidation is seen in the biosynthesis of GE37468, in which Ile8 is transformed into a b-methyl-d-hydroxy-proline residue via the actions of P450 GetJ. This reaction would appear to follow the same path as is generally observed for the oxidation of methyl groups into carboxylic acids, although in this case the intermediate aldehyde spontaneously cyclises to generate a hemiaminal.…”

Section: Peptide Biosynthesis Pathwaysmentioning

confidence: 93%

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

et al. 2018

View full text Add to dashboard Cite

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.

show abstract

Section: Peptide Biosynthesis Pathwaysmentioning

confidence: 93%

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

et al. 2018

View full text Add to dashboard Cite

show abstract

“…182 Similarly, the thiomuracin cyclodehydratase (section 3.2.4) has a non-linear preferred order but only for first three cyclodehydrations, as the remaining thiazolines are installed without detectable intermediates or order. 632 …”

Section: Characterization Of Ripp Cyclodehydratasesmentioning

confidence: 99%

“…361 On the other hand, the thiomuracin cyclodehydratase also has no difficulty in processing around altered Cys sites. 632 A molecular level understanding of these preferences has yet to be determined.…”

Section: Characterization Of Ripp Cyclodehydratasesmentioning

confidence: 99%

YcaO-Dependent Posttranslational Amide Activation: Biosynthesis, Structure, and Function

et al. 2017

Self Cite

View full text Add to dashboard Cite

With advances in sequencing technology, uncharacterized proteins and domains of unknown function (DUFs) are rapidly accumulating in sequence databases and offer an opportunity to discover new protein chemistry and reaction mechanisms. The focus of this review, the formerly enigmatic YcaO superfamily (DUF181), has been found to catalyze a unique phosphorylation of a ribosomal peptide backbone amide upon attack by different nucleophiles. Established nucleophiles are the side chains of Cys, Ser, and Thr which gives rise to azoline/azole biosynthesis in ribosomally synthesized and posttranslationally modified peptide (RiPP) natural products. However, much remains unknown about the potential for YcaO proteins to collaborate with other nucleophiles. Recent work suggests potential in forming thioamides, macroamidines, and possibly additional post-translational modifications. This review covers all knowledge through mid-2016 regarding the biosynthetic gene clusters (BGCs), natural products, functions, mechanisms, and applications of YcaO proteins and outlines likely future research directions for this protein superfamily.

show abstract

“…1 Although TbtI lacks an identifiable RiPP Recognition Element (RRE), 36 an indicator of LP dependence, TbtD also lacks an RRE yet is LP-dependent; 7 further, the RRE-containing TbtB is LP-independent. 8 To assess the LP-dependence of TbtI, 2 was treated with the endoproteinase GluC to generate 7 , which retains Val-Gly-Ala from the LP. In testing 7 as a TbtI substrate, we observed full conversion to the methylthiazole 8 (Figure 2).…”

mentioning

confidence: 99%

“…37,38 To investigate whether Asn3 governs the regioselectivity of TbtI, we replaced this residue in TbtA with Ala, Asp, and Gln, followed by hexazole production in E. coli . 8 The N3A-, N3D- and N3Q-bearing variants of 7 were then anaerobically treated with TbtI, prior to MALDI-TOF-MS analysis. Under the conditions used, the reaction with substrate 7 went to completion.…”

mentioning

confidence: 99%

Reconstitution and Substrate Specificity of the Radical S-Adenosyl-methionine Thiazole C-Methyltransferase in Thiomuracin Biosynthesis

et al. 2017

Self Cite

View full text Add to dashboard Cite

Thiomuracin is a thiopeptide antibiotic with potent activity towards Gram-positive drug-resistant bacteria. Thiomuracin is biosynthesized from a precursor peptide, TbtA, by a complex array of posttranslational modifications. One of several intriguing transformations is the C-methylation of thiazole, occurring at an unactivated sp2 carbon. Herein, we report the in vitro reconstitution of TbtI, the responsible radical S-adenosyl-methionine (rSAM) C-methyltransferase, which catalyzes the formation of 5-methylthiazole at a single site. Our studies demonstrate that a linear hexazole-bearing intermediate of TbtA is a substrate for TbtI whereas macrocyclized thiomuracin GZ is not. In determining the minimal substrate for TbtI, we found that the enzyme is functional when most of the leader peptide has been removed. The in vitro reconstitution of TbtI, a class C rSAM methyltransferase, further adds to the chemical versatility of rSAM enzymes, and informs on the complexity of thiomuracin biosynthesis.

show abstract

Biosynthetic Timing and Substrate Specificity for the Thiopeptide Thiomuracin

Cited by 77 publications

References 18 publications

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

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

YcaO-Dependent Posttranslational Amide Activation: Biosynthesis, Structure, and Function

Reconstitution and Substrate Specificity of the Radical S-Adenosyl-methionine Thiazole C-Methyltransferase in Thiomuracin Biosynthesis

Contact Info

Product

Resources

About