Mechanistic insights into class B radical-S-adenosylmethionine methylases: ubiquitous tailoring enzymes in natural product biosynthesis

Zhou, Shanshan; Alkhalaf, Lona M.; Santos, Emmanuel L. C. de los; Challis, Gregory L.

doi:10.1016/j.cbpa.2016.08.021

Cited by 35 publications

(34 citation statements)

References 42 publications

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“…Such enzymes are known to catalyse the methylation of unactivated carbon centres in the biosynthesis of a variety of specialised metabolites. 22 Taken together, these analyses suggested that the sven0503 – sven0517 gene cluster may direct the biosynthesis of a C-methylated pyochelin derivative.…”

Section: Resultsmentioning

confidence: 92%

“…Type B radical-SAM methylases are known to catalyse methylation of unactivated carbon centres in the biosynthesis of several different classes of natural products, including aminoglycosides, β-lactams, phosphonates, and ribosomally biosynthesised and posttranslationally-modified peptides. 22 However, to the best of our knowledge the Sven0515-catalysed methylation of thiazostatin 6 is the first experimentally-validated example of such a reaction in the biosynthesis of a nonribosomal peptide.…”

Section: Discussionmentioning

confidence: 97%

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Watasemycin biosynthesis in Streptomyces venezuelae: thiazoline C-methylation by a type B radical-SAM methylase homologue

et al. 2017

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Section: Resultsmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 97%

Watasemycin biosynthesis in Streptomyces venezuelae: thiazoline C-methylation by a type B radical-SAM methylase homologue

et al. 2017

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“…21 These findings led to the suggestion that Me-Cbl and the radical SAM machinery are situated on opposite sides of the center to be methylated once bound in the active site of these enzymes. 9,11,21 This hypothesis is consistent with the only known crystal structure of a Cbl-dependent radical SAM enzyme, OxsB. 25 Though not a RSMT, the substrate binding site of OxsB is postulated to be interposed equidistant between the C5′ of SAM and the Co of the Cbl cofactor.…”

mentioning

confidence: 99%

Reaction Catalyzed by GenK, a Cobalamin-Dependent Radical S-Adenosyl-l-methionine Methyltransferase in the Biosynthetic Pathway of Gentamicin, Proceeds with Retention of Configuration

et al. 2017

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Many cobalamin (Cbl)-dependent radical S-adenosyl-L-methionine (SAM) methyltransferases have been identified through sequence alignment and/or genetic analysis; however, few have been studied in vitro. GenK is one such enzyme that catalyzes methylation of the 6′-carbon of gentamicin X2 (GenX2) to produce G418 during the biosynthesis of gentamicins. Reported herein, several alternative substrates and fluorinated substrate analogs were prepared to investigate the mechanism of methyl transfer from Cbl to the substrate as well as the substrate specificity of GenK. Experiments with deuterated substrates are also shown here to demonstrate that the 6′-pro-R-hydrogen atom of GenX2 is stereoselectively abstracted by the 5′-dAdo· radical and that methylation occurs with retention of configuration at C6′. Based on these observations, a model of GenK catalysis is proposed wherein free rotation of the radical-bearing carbon is prevented and the radical SAM machinery sits adjacent rather than opposite to the Me-Cbl cofactor with respect to the substrate in the enzyme active site.

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“…GenD2 and GenS are responsible for the amination of the C3'' position of gentamicin A2, and the resulting amino group is methylated by GenN to afford a C3'' methylamino group 28 . GenD1 belongs to the class B radical SAM methyltransferases 29, 30 , which utilizes a cobalamin cofactor and installs a methyl group on the C4'' position 31, 32 . Gentamicin X2 is then methylated at the C6' position by GenK, which, like GenD1, also belongs to the class B radical SAM methyltransferase, leading to the production of gentamicin G418 33 .…”

Section: Parallel Pathways In Gentamicin Biosynthesismentioning

confidence: 99%

Parallel pathways in the biosynthesis of aminoglycoside antibiotics

Zhang

Deng

2017

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Despite their inherent toxicity and the global spread of bacterial resistance, aminoglycosides (AGs), an old class of microbial drugs, remain a valuable component of the antibiotic arsenal. Recent studies have continued to reveal the fascinating biochemistry of AG biosynthesis and the rich potential in their pathway engineering. In particular, parallel pathways have been shown to be common and widespread in AG biosynthesis, highlighting nature’s ingenuity in accessing diverse natural products from a limited set of genes. In this review, we discuss the parallel biosynthetic pathways of three representative AG antibiotics—kanamycin, gentamicin, and apramycin—as well as future directions towards the discovery and development of novel AGs.

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Mechanistic insights into class B radical-S-adenosylmethionine methylases: ubiquitous tailoring enzymes in natural product biosynthesis

Cited by 35 publications

References 42 publications

Watasemycin biosynthesis in Streptomyces venezuelae: thiazoline C-methylation by a type B radical-SAM methylase homologue

Watasemycin biosynthesis in Streptomyces venezuelae: thiazoline C-methylation by a type B radical-SAM methylase homologue

Reaction Catalyzed by GenK, a Cobalamin-Dependent Radical S-Adenosyl-l-methionine Methyltransferase in the Biosynthetic Pathway of Gentamicin, Proceeds with Retention of Configuration

Parallel pathways in the biosynthesis of aminoglycoside antibiotics

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