Ketoreductase domain dysfunction expands chemodiversity: malyngamide biosynthesis in the cyanobacterium Okeania hirsuta

Nathan Moss, Tiago Leao, Michael Rankin, Tyler M. McCullough, Pingping Qu, Антон Иванович Коробейников, Janet L. Smith, Lena Gerwick, William H. Gerwick

Результат исследований: Научные публикации в периодических изданияхстатья

2 Цитирования (Scopus)

Выдержка

Dozens of type A malyngamides, principally identified by a decorated six-membered cyclohexanone head group and methoxylated lyngbic acid tail, have been isolated over several decades. Their environmental sources include macro- and microbiotic organisms, including sea hares, red alga, and cyanobacterial assemblages but their true producing organism has remained enigmatic. Many type A analogs display potent bioactivity in human-health related assays, spurring an interest in this molecular class and its biosynthetic pathway. Here we present the discovery of the type A malyngamide biosynthetic pathway in the first sequenced genome of the cyanobacterial genus Okeania. Bioinformatic analysis of two cultured Okeania genome assemblies identified 62 and 68 kb polyketide synthase/non-ribosomal peptide synthetase (PKS/NRPS) pathways with unusual loading and termination genes. NMR data of malyngamide C acetate derived from 13C-substrate-fed cultures provided evidence that an intact octanoate moiety is transferred to the first KS module via a LipM homolog originally associated with lipoic acid metabolism and implicated an inactive ketoreductase (KR0) as critical for six-membered ring formation, a hallmark of the malyngamide family. Phylogenetic analysis and homology modeling of the penultimate KR0 domain inferred structural cofactor-binding and active site alterations as contributory to domain dysfunction, which was confirmed by recombinant protein expression and NADPH binding assay. The carbonyl retained from this KR0 ultimately enables an intramolecular Knoevenagel condensation to form the characteristic cyclohexanone ring. Understanding this critical step allows assignment of a biosynthetic model for all type A malyngamides, whereby well-characterized tailoring modifications explain the surprising proliferation and diversity of analogs.
Язык оригиналаанглийский
ЖурналACS Chemical Biology
DOI
СостояниеЭлектронная публикация перед печатью - 16 ноя 2018

Отпечаток

Biosynthesis
Biosynthetic Pathways
Cyanobacteria
Genes
Peptide Synthases
Genome
Polyketide Synthases
Hares
Rhodophyta
Thioctic Acid
Assays
Computational Biology
NADP
Recombinant Proteins
Oceans and Seas
Tail
Catalytic Domain
Acetates
Binding Sites
Bioinformatics

Цитировать

Moss, Nathan ; Leao, Tiago ; Rankin, Michael ; McCullough, Tyler M. ; Qu, Pingping ; Коробейников, Антон Иванович ; Smith, Janet L. ; Gerwick, Lena ; Gerwick, William H. / Ketoreductase domain dysfunction expands chemodiversity: malyngamide biosynthesis in the cyanobacterium Okeania hirsuta. В: ACS Chemical Biology. 2018.
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title = "Ketoreductase domain dysfunction expands chemodiversity: malyngamide biosynthesis in the cyanobacterium Okeania hirsuta",
abstract = "Dozens of type A malyngamides, principally identified by a decorated six-membered cyclohexanone head group and methoxylated lyngbic acid tail, have been isolated over several decades. Their environmental sources include macro- and microbiotic organisms, including sea hares, red alga, and cyanobacterial assemblages but their true producing organism has remained enigmatic. Many type A analogs display potent bioactivity in human-health related assays, spurring an interest in this molecular class and its biosynthetic pathway. Here we present the discovery of the type A malyngamide biosynthetic pathway in the first sequenced genome of the cyanobacterial genus Okeania. Bioinformatic analysis of two cultured Okeania genome assemblies identified 62 and 68 kb polyketide synthase/non-ribosomal peptide synthetase (PKS/NRPS) pathways with unusual loading and termination genes. NMR data of malyngamide C acetate derived from 13C-substrate-fed cultures provided evidence that an intact octanoate moiety is transferred to the first KS module via a LipM homolog originally associated with lipoic acid metabolism and implicated an inactive ketoreductase (KR0) as critical for six-membered ring formation, a hallmark of the malyngamide family. Phylogenetic analysis and homology modeling of the penultimate KR0 domain inferred structural cofactor-binding and active site alterations as contributory to domain dysfunction, which was confirmed by recombinant protein expression and NADPH binding assay. The carbonyl retained from this KR0 ultimately enables an intramolecular Knoevenagel condensation to form the characteristic cyclohexanone ring. Understanding this critical step allows assignment of a biosynthetic model for all type A malyngamides, whereby well-characterized tailoring modifications explain the surprising proliferation and diversity of analogs.",
author = "Nathan Moss and Tiago Leao and Michael Rankin and McCullough, {Tyler M.} and Pingping Qu and Коробейников, {Антон Иванович} and Smith, {Janet L.} and Lena Gerwick and Gerwick, {William H.}",
year = "2018",
month = "11",
day = "16",
doi = "10.1021/acschembio.8b00910",
language = "English",
journal = "ACS Chemical Biology",
issn = "1554-8929",
publisher = "American Chemical Society",

}

Ketoreductase domain dysfunction expands chemodiversity: malyngamide biosynthesis in the cyanobacterium Okeania hirsuta. / Moss, Nathan; Leao, Tiago; Rankin, Michael; McCullough, Tyler M.; Qu, Pingping; Коробейников, Антон Иванович; Smith, Janet L.; Gerwick, Lena; Gerwick, William H.

В: ACS Chemical Biology, 16.11.2018.

Результат исследований: Научные публикации в периодических изданияхстатья

TY - JOUR

T1 - Ketoreductase domain dysfunction expands chemodiversity: malyngamide biosynthesis in the cyanobacterium Okeania hirsuta

AU - Moss, Nathan

AU - Leao, Tiago

AU - Rankin, Michael

AU - McCullough, Tyler M.

AU - Qu, Pingping

AU - Коробейников, Антон Иванович

AU - Smith, Janet L.

AU - Gerwick, Lena

AU - Gerwick, William H.

PY - 2018/11/16

Y1 - 2018/11/16

N2 - Dozens of type A malyngamides, principally identified by a decorated six-membered cyclohexanone head group and methoxylated lyngbic acid tail, have been isolated over several decades. Their environmental sources include macro- and microbiotic organisms, including sea hares, red alga, and cyanobacterial assemblages but their true producing organism has remained enigmatic. Many type A analogs display potent bioactivity in human-health related assays, spurring an interest in this molecular class and its biosynthetic pathway. Here we present the discovery of the type A malyngamide biosynthetic pathway in the first sequenced genome of the cyanobacterial genus Okeania. Bioinformatic analysis of two cultured Okeania genome assemblies identified 62 and 68 kb polyketide synthase/non-ribosomal peptide synthetase (PKS/NRPS) pathways with unusual loading and termination genes. NMR data of malyngamide C acetate derived from 13C-substrate-fed cultures provided evidence that an intact octanoate moiety is transferred to the first KS module via a LipM homolog originally associated with lipoic acid metabolism and implicated an inactive ketoreductase (KR0) as critical for six-membered ring formation, a hallmark of the malyngamide family. Phylogenetic analysis and homology modeling of the penultimate KR0 domain inferred structural cofactor-binding and active site alterations as contributory to domain dysfunction, which was confirmed by recombinant protein expression and NADPH binding assay. The carbonyl retained from this KR0 ultimately enables an intramolecular Knoevenagel condensation to form the characteristic cyclohexanone ring. Understanding this critical step allows assignment of a biosynthetic model for all type A malyngamides, whereby well-characterized tailoring modifications explain the surprising proliferation and diversity of analogs.

AB - Dozens of type A malyngamides, principally identified by a decorated six-membered cyclohexanone head group and methoxylated lyngbic acid tail, have been isolated over several decades. Their environmental sources include macro- and microbiotic organisms, including sea hares, red alga, and cyanobacterial assemblages but their true producing organism has remained enigmatic. Many type A analogs display potent bioactivity in human-health related assays, spurring an interest in this molecular class and its biosynthetic pathway. Here we present the discovery of the type A malyngamide biosynthetic pathway in the first sequenced genome of the cyanobacterial genus Okeania. Bioinformatic analysis of two cultured Okeania genome assemblies identified 62 and 68 kb polyketide synthase/non-ribosomal peptide synthetase (PKS/NRPS) pathways with unusual loading and termination genes. NMR data of malyngamide C acetate derived from 13C-substrate-fed cultures provided evidence that an intact octanoate moiety is transferred to the first KS module via a LipM homolog originally associated with lipoic acid metabolism and implicated an inactive ketoreductase (KR0) as critical for six-membered ring formation, a hallmark of the malyngamide family. Phylogenetic analysis and homology modeling of the penultimate KR0 domain inferred structural cofactor-binding and active site alterations as contributory to domain dysfunction, which was confirmed by recombinant protein expression and NADPH binding assay. The carbonyl retained from this KR0 ultimately enables an intramolecular Knoevenagel condensation to form the characteristic cyclohexanone ring. Understanding this critical step allows assignment of a biosynthetic model for all type A malyngamides, whereby well-characterized tailoring modifications explain the surprising proliferation and diversity of analogs.

U2 - 10.1021/acschembio.8b00910

DO - 10.1021/acschembio.8b00910

M3 - Article

JO - ACS Chemical Biology

JF - ACS Chemical Biology

SN - 1554-8929

ER -