Biosynthesis of the antituberculous agent caprazamycin: Identification of caprazol-3ʺ-phosphate, an unprecedented caprazamycin-related metabolite

Shiraishi, Taro; Hiro, Noboru; Igarashi, Masayuki; Nishiyama, Makoto; Kuzuyama, Tomohisa

doi:10.2323/jgam.2016.01.002

Cited by 10 publications

(12 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mur28 kinase from the muraymycin pathway catalyzes the 3”-OH phosphorylation of a GlyU-ADR disaccharide biosynthetic intermediate 24 , and homologous kinases are conserved among GlyU-ADR containing nucleoside BGCs (caprazamycin 25 , A-90289 26 , and muraminomycin 27 ). A 3”-phosphorylated caprazamycin precursor has been isolated 28 . More recently, while this manuscript was in revision, one of the muraymycin biosynthetic enzymes (Mur24) was reported to act specifically on 3”-phosphorylated GlyU-ADR 29 .…”

Section: Resultsmentioning

confidence: 99%

Cryptic phosphorylation in nucleoside natural product biosynthesis

et al. 2020

View full text Add to dashboard Cite

Kinases are annotated in many nucleoside biosynthetic gene clusters (BGCs) but generally are considered responsible only for self-resistance. Here, we report an unexpected 2’-phosphorylation of nucleoside biosynthetic intermediates in the nikkomycin and polyoxin pathways. This phosphorylation is a unique cryptic modification as it is introduced in the third of seven steps during aminohexuronic acid (AHA) nucleoside biosynthesis, retained throughout the pathway’s duration, and is removed in the last step of the pathway. Bioinformatic analysis of reported nucleoside BGCs suggests the presence of cryptic phosphorylation in other pathways and the importance of functional characterization of kinases in nucleoside biosynthetic pathways in general. This study also functionally characterized all of the enzymes responsible for AHA biosynthesis and revealed that AHA is constructed via a unique oxidative C-C bond cleavage reaction. The results suggest a divergent biosynthetic mechanism for three classes of antifungal nucleoside natural products.

show abstract

Section: Resultsmentioning

confidence: 99%

Cryptic phosphorylation in nucleoside natural product biosynthesis

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Thus, the data are consistent with Mur28 catalysis occurring prior to the attachment of the aminopropyl-linked peptide. These conclusions are supported by the discovery of phosphorylated caprazol from a caprazamycin-producing strain (11), suggesting that phosphorylation occurs prior to complete fabrication of A90289-type antibiotics.…”

Section: Discussionmentioning

confidence: 80%

“…Recently, a new muraymycin (C6) acetylated at the 5Љ-amine of the ADR moiety, with modestly reduced (Ն6-fold) IC 50 values against recombinant MraYs from different sources, was isolated (10). In a separate report, a 3Љ-O-phosphorylated caprazamycin precursor called caprazol-3Љ-phosphate was unexpectedly isolated following inactivation of cpz23, encoding a putative SGNH hydrolase, although in that case the inhibitory or antibacterial activity of the phosphorylated variant was not tested (11). Covalent modifications of these types (phosphorylation and acetylation), along with nucleotidylation, are often found as mechanisms of resistance to antibiotics, most notably aminoglycosides (12)(13)(14)(15).…”

mentioning

confidence: 99%

Self-Resistance during Muraymycin Biosynthesis: a Complementary Nucleotidyltransferase and Phosphotransferase with Identical Modification Sites and Distinct Temporal Order

Cui

Wang

Liu

et al. 2018

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Muraymycins are antibacterial natural products from spp. that inhibit translocase I (MraY), which is involved in cell wall biosynthesis. Structurally, muraymycins consist of a 5'--glycyluridine (GlyU) appended to a 5″-amino-5″-deoxyribose (ADR), forming a disaccharide core that is found in several peptidyl nucleoside inhibitors of MraY. For muraymycins, the GlyU-ADR disaccharide is further modified with an aminopropyl-linked peptide to generate the simplest structures, annotated as the muraymycin D series. Two enzymes encoded in the muraymycin biosynthetic gene cluster, Mur29 and Mur28, were functionally assigned as a Mg·ATP-dependent nucleotidyltransferase and a Mg·ATP-dependent phosphotransferase, respectively, both modifying the 3″-OH of the disaccharide. Biochemical characterization revealed that both enzymes can utilize several nucleotide donors as cosubstrates and the acceptor substrate muraymycin also behaves as an inhibitor. Single-substrate kinetic analyses revealed that Mur28 preferentially phosphorylates a synthetic GlyU-ADR disaccharide, a hypothetical biosynthetic precursor of muraymycins, while Mur29 preferentially adenylates the D series of muraymycins. The adenylated or phosphorylated products have significantly reduced (170-fold and 51-fold, respectively) MraY inhibitory activities and reduced antibacterial activities, compared with the respective unmodified muraymycins. The results are consistent with Mur29-catalyzed adenylation and Mur28-catalyzed phosphorylation serving as complementary self-resistance mechanisms, with a distinct temporal order during muraymycin biosynthesis.

show abstract

“…The proposed mechanism involves phosphorylation of 2′-OH of a precursor nucleotide early in the pathway, which is essential for the activity of all subsequent AHA biosynthetic enzymes. Analogous cryptic phosphorylation has also been found in other nucleoside − and aminoglycoside natural products, suggesting the generality of this biosynthetic mechanism.…”

mentioning

confidence: 55%

Conserved Mechanism of 2′-Phosphorylation-Aided Amide Ligation in Peptidyl Nucleoside Biosynthesis

2021

View full text Add to dashboard Cite

Peptidyl nucleoside antifungals, represented by nikkomycins and polyoxins, consist of an unusual six-carbon nucleoside [aminohexuronic acid (AHA)] ligated to a nonproteinogenic amino acid via an amide bond. A recent study suggested that AHA is biosynthesized through cryptic phosphorylation, where a 2′-phosphate is introduced early in the pathway and required to form AHA. However, whether 2′-phosphorylation is necessary for the last step of biosynthesis, the formation of the amide bond between AHA and nonproteinogenic amino acids, remains ambiguous. Here, we address this question with comprehensive in vitro and in vivo characterizations of PolG and NikS, which together provide strong evidence that amide ligation proceeds with 2′phosphorylated substrates in both pathways. Our results suggest that 2′-phosphorylation is retained for the entirety of both nikkomycin and polyoxin biosynthesis, providing important insights into how cryptic phosphorylation assists with nucleoside natural product biosynthesis. P eptidyl nucleosides (PNs), such as nikkomycins and polyoxins, exhibit selective antifungal activities with no known side effects to higher eukaryotes. Consequently, polyoxin D [1 (Figure 1A)] is a safely used fungicide in Communication pubs.acs.org/biochemistry

show abstract

Biosynthesis of the antituberculous agent caprazamycin: Identification of caprazol-3ʺ-phosphate, an unprecedented caprazamycin-related metabolite

Cited by 10 publications

References 13 publications

Cryptic phosphorylation in nucleoside natural product biosynthesis

Cryptic phosphorylation in nucleoside natural product biosynthesis

Self-Resistance during Muraymycin Biosynthesis: a Complementary Nucleotidyltransferase and Phosphotransferase with Identical Modification Sites and Distinct Temporal Order

Conserved Mechanism of 2′-Phosphorylation-Aided Amide Ligation in Peptidyl Nucleoside Biosynthesis

Contact Info

Product

Resources

About