Identification and characterization of the echinocandin b biosynthetic gene cluster from Emericella rugulosa NRRL 11440

R.A. Cacho, W. Jiang, Heng Chooi, C.T. Walsh, Y. Tang

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Echinocandins are a family of fungal lipidated cyclic hexapeptide natural products. Due to their effectiveness as antifungal agents, three semisynthetic derivatives have been developed and approved for treatment of human invasive candidiasis. All six of the amino acid residues are hydroxylated, including 4R,5R-dihydroxy-l-ornithine, 4R-hydroxyl-l-proline, 3S,4S-dihydroxy-l- homotyrosine, and 3S-hydroxyl-4S-methyl-l-proline. We report here the biosynthetic gene cluster of echinocandin B 1 from Emericella rugulosa NRRL 11440 containing genes encoding for a six-module nonribosomal peptide synthetase EcdA, an acyl-AMP ligase EcdI, and oxygenases EcdG, EcdH, and EcdK. We showed EcdI activates linoleate as linoleyl-AMP and installs it on the first thiolation domain of EcdA. We have also established through ATP-PPi exchange assay that EcdA loads l-ornithine in the first module. A separate hty gene cluster encodes four enzymes for de novo generation of l-homotyrosine from acetyl-CoA and 4-hydroxyphenyl-pyruvate is found from the sequenced genome. Deletions in the ecdA, and htyA genes validate their essential roles in echinocandin B production. Five predicted iron-centered oxygenase genes, ecdG, ecdH, ecdK, htyE, and htyF, in the two separate ecd and hty clusters are likely to be the tailoring oxygenases for maturation of the nascent NRPS lipohexapeptidolactam product. © 2012 American Chemical Society.
Original languageEnglish
Pages (from-to)16781-16790
JournalJournal of the American Chemical Society
Volume134
Issue number40
DOIs
Publication statusPublished - 2012

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Emericella
Echinocandins
Oxygenases
Multigene Family
Ornithine
Genes
Adenosine Monophosphate
Proline
Hydroxyl Radical
Invasive Candidiasis
Peptide Synthases
Acetyl Coenzyme A
Antifungal Agents
Linoleic Acid
Ligases
Biological Products
Pyruvic Acid
Antifungal agents
Iron
Adenosine Triphosphate

Cite this

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abstract = "Echinocandins are a family of fungal lipidated cyclic hexapeptide natural products. Due to their effectiveness as antifungal agents, three semisynthetic derivatives have been developed and approved for treatment of human invasive candidiasis. All six of the amino acid residues are hydroxylated, including 4R,5R-dihydroxy-l-ornithine, 4R-hydroxyl-l-proline, 3S,4S-dihydroxy-l- homotyrosine, and 3S-hydroxyl-4S-methyl-l-proline. We report here the biosynthetic gene cluster of echinocandin B 1 from Emericella rugulosa NRRL 11440 containing genes encoding for a six-module nonribosomal peptide synthetase EcdA, an acyl-AMP ligase EcdI, and oxygenases EcdG, EcdH, and EcdK. We showed EcdI activates linoleate as linoleyl-AMP and installs it on the first thiolation domain of EcdA. We have also established through ATP-PPi exchange assay that EcdA loads l-ornithine in the first module. A separate hty gene cluster encodes four enzymes for de novo generation of l-homotyrosine from acetyl-CoA and 4-hydroxyphenyl-pyruvate is found from the sequenced genome. Deletions in the ecdA, and htyA genes validate their essential roles in echinocandin B production. Five predicted iron-centered oxygenase genes, ecdG, ecdH, ecdK, htyE, and htyF, in the two separate ecd and hty clusters are likely to be the tailoring oxygenases for maturation of the nascent NRPS lipohexapeptidolactam product. {\circledC} 2012 American Chemical Society.",
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Identification and characterization of the echinocandin b biosynthetic gene cluster from Emericella rugulosa NRRL 11440. / Cacho, R.A.; Jiang, W.; Chooi, Heng; Walsh, C.T.; Tang, Y.

In: Journal of the American Chemical Society, Vol. 134, No. 40, 2012, p. 16781-16790.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification and characterization of the echinocandin b biosynthetic gene cluster from Emericella rugulosa NRRL 11440

AU - Cacho, R.A.

AU - Jiang, W.

AU - Chooi, Heng

AU - Walsh, C.T.

AU - Tang, Y.

PY - 2012

Y1 - 2012

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AB - Echinocandins are a family of fungal lipidated cyclic hexapeptide natural products. Due to their effectiveness as antifungal agents, three semisynthetic derivatives have been developed and approved for treatment of human invasive candidiasis. All six of the amino acid residues are hydroxylated, including 4R,5R-dihydroxy-l-ornithine, 4R-hydroxyl-l-proline, 3S,4S-dihydroxy-l- homotyrosine, and 3S-hydroxyl-4S-methyl-l-proline. We report here the biosynthetic gene cluster of echinocandin B 1 from Emericella rugulosa NRRL 11440 containing genes encoding for a six-module nonribosomal peptide synthetase EcdA, an acyl-AMP ligase EcdI, and oxygenases EcdG, EcdH, and EcdK. We showed EcdI activates linoleate as linoleyl-AMP and installs it on the first thiolation domain of EcdA. We have also established through ATP-PPi exchange assay that EcdA loads l-ornithine in the first module. A separate hty gene cluster encodes four enzymes for de novo generation of l-homotyrosine from acetyl-CoA and 4-hydroxyphenyl-pyruvate is found from the sequenced genome. Deletions in the ecdA, and htyA genes validate their essential roles in echinocandin B production. Five predicted iron-centered oxygenase genes, ecdG, ecdH, ecdK, htyE, and htyF, in the two separate ecd and hty clusters are likely to be the tailoring oxygenases for maturation of the nascent NRPS lipohexapeptidolactam product. © 2012 American Chemical Society.

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M3 - Article

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SP - 16781

EP - 16790

JO - Journal of the Amercian Chemical Society

JF - Journal of the Amercian Chemical Society

SN - 0002-7863

IS - 40

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