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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
ACS chemical biology
2024
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/39485010/ |
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| author | Chaudhri, Ayesha Ahmed Kakumu, Yuya Thiengmag, Sirinthra Liu, Jack Chun-Ting Lin, Geng-Min Durusu, Suhan Biermann, Friederike Boeck, Miriam Voigt, Christopher A Clardy, Jon Ueoka, Reiko Walker, Allison S Helfrich, Eric J N |
| author_facet | Chaudhri, Ayesha Ahmed Kakumu, Yuya Thiengmag, Sirinthra Liu, Jack Chun-Ting Lin, Geng-Min Durusu, Suhan Biermann, Friederike Boeck, Miriam Voigt, Christopher A Clardy, Jon Ueoka, Reiko Walker, Allison S Helfrich, Eric J N Chaudhri, Ayesha Ahmed Kakumu, Yuya Thiengmag, Sirinthra Liu, Jack Chun-Ting Lin, Geng-Min Durusu, Suhan Biermann, Friederike Boeck, Miriam Voigt, Christopher A Clardy, Jon Ueoka, Reiko Walker, Allison S Helfrich, Eric J N |
| collection | PubMed - marine biology |
| contents | Functional Redundancy and Dual Function of a Hypothetical Protein in the Biosynthesis of Eunicellane-Type Diterpenoids. Chaudhri, Ayesha Ahmed Kakumu, Yuya Thiengmag, Sirinthra Liu, Jack Chun-Ting Lin, Geng-Min Durusu, Suhan Biermann, Friederike Boeck, Miriam Voigt, Christopher A Clardy, Jon Ueoka, Reiko Walker, Allison S Helfrich, Eric J N Diterpenes Cytochrome P-450 Enzyme System Multigene Family Bacterial Proteins Actinobacteria Biosynthetic Pathways Many complex terpenoids, predominantly isolated from plants and fungi, show drug-like physicochemical properties. Recent advances in genome mining revealed actinobacteria as an almost untouched treasure trove of terpene biosynthetic gene clusters (BGCs). In this study, we characterized a terpene BGC with an unusual architecture. The selected BGC includes, among others, genes encoding a terpene cyclase fused to a truncated reductase domain and a cytochrome P450 monooxygenase (P450) that is split over three gene fragments. Functional characterization of the BGC in a heterologous host led to the identification of several new members of the -eunicellane family of diterpenoids, the euthailols, that feature unique oxidation patterns. A combination of bioinformatic analyses, structural modeling studies, and heterologous expression revealed a dual function of the pathway-encoded hypothetical protein that acts as an isomerase and an oxygenase. Moreover, in the absence of other tailoring enzymes, a P450 hydroxylates the eunicellane scaffold at a position that is not modified in other eunicellanes. Surprisingly, both the modifications installed by the hypothetical protein and one of the P450s exhibit partial redundancy. Bioactivity assays revealed that some of the euthailols show growth inhibitory properties against Gram-negative ESKAPE pathogens. The characterization of the euthailol BGC in this study provides unprecedented insights into the partial functional redundancy of tailoring enzymes in complex diterpenoid biosynthesis and highlights hypothetical proteins as an important and largely overlooked family of tailoring enzymes involved in the maturation of complex terpenoids. |
| format | Artículo científico |
| id | pubmed_39485010 |
| institution | PubMed |
| language | en |
| publishDate | 2024 |
| publisher | ACS chemical biology |
| record_format | pubmed |
| spellingShingle | Functional Redundancy and Dual Function of a Hypothetical Protein in the Biosynthesis of Eunicellane-Type Diterpenoids. Chaudhri, Ayesha Ahmed Kakumu, Yuya Thiengmag, Sirinthra Liu, Jack Chun-Ting Lin, Geng-Min Durusu, Suhan Biermann, Friederike Boeck, Miriam Voigt, Christopher A Clardy, Jon Ueoka, Reiko Walker, Allison S Helfrich, Eric J N Diterpenes Cytochrome P-450 Enzyme System Multigene Family Bacterial Proteins Actinobacteria Biosynthetic Pathways Functional Redundancy and Dual Function of a Hypothetical Protein in the Biosynthesis of Eunicellane-Type Diterpenoids. Chaudhri, Ayesha Ahmed Kakumu, Yuya Thiengmag, Sirinthra Liu, Jack Chun-Ting Lin, Geng-Min Durusu, Suhan Biermann, Friederike Boeck, Miriam Voigt, Christopher A Clardy, Jon Ueoka, Reiko Walker, Allison S Helfrich, Eric J N Diterpenes Cytochrome P-450 Enzyme System Multigene Family Bacterial Proteins Actinobacteria Biosynthetic Pathways Many complex terpenoids, predominantly isolated from plants and fungi, show drug-like physicochemical properties. Recent advances in genome mining revealed actinobacteria as an almost untouched treasure trove of terpene biosynthetic gene clusters (BGCs). In this study, we characterized a terpene BGC with an unusual architecture. The selected BGC includes, among others, genes encoding a terpene cyclase fused to a truncated reductase domain and a cytochrome P450 monooxygenase (P450) that is split over three gene fragments. Functional characterization of the BGC in a heterologous host led to the identification of several new members of the -eunicellane family of diterpenoids, the euthailols, that feature unique oxidation patterns. A combination of bioinformatic analyses, structural modeling studies, and heterologous expression revealed a dual function of the pathway-encoded hypothetical protein that acts as an isomerase and an oxygenase. Moreover, in the absence of other tailoring enzymes, a P450 hydroxylates the eunicellane scaffold at a position that is not modified in other eunicellanes. Surprisingly, both the modifications installed by the hypothetical protein and one of the P450s exhibit partial redundancy. Bioactivity assays revealed that some of the euthailols show growth inhibitory properties against Gram-negative ESKAPE pathogens. The characterization of the euthailol BGC in this study provides unprecedented insights into the partial functional redundancy of tailoring enzymes in complex diterpenoid biosynthesis and highlights hypothetical proteins as an important and largely overlooked family of tailoring enzymes involved in the maturation of complex terpenoids. |
| title | Functional Redundancy and Dual Function of a Hypothetical Protein in the Biosynthesis of Eunicellane-Type Diterpenoids. |
| topic | Diterpenes Cytochrome P-450 Enzyme System Multigene Family Bacterial Proteins Actinobacteria Biosynthetic Pathways |
| url | https://pubmed.ncbi.nlm.nih.gov/39485010/ |