دورية أكاديمية
أعرض في EDS

Heterologous expression of the atypical tetracycline chelocardin reveals the full set of genes required for its biosynthesis.

التفاصيل البيبلوغرافية
العنوان: Heterologous expression of the atypical tetracycline chelocardin reveals the full set of genes required for its biosynthesis.
المؤلفون: Lukežič T; Department of Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)-Helmholtz Centre for Infection Research (HZI), and Department of Pharmacy, Saarland University Campus, Campus E8.1, 66123, Saarbrücken, Germany.; German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, 38124, Braunschweig, Germany.; National Institute of Biology, Večna pot 111, 1000, Ljubljana, Slovenia., Pikl Š; Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia., Zaburannyi N; Department of Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)-Helmholtz Centre for Infection Research (HZI), and Department of Pharmacy, Saarland University Campus, Campus E8.1, 66123, Saarbrücken, Germany.; German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, 38124, Braunschweig, Germany., Remškar M; Department of Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)-Helmholtz Centre for Infection Research (HZI), and Department of Pharmacy, Saarland University Campus, Campus E8.1, 66123, Saarbrücken, Germany.; German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, 38124, Braunschweig, Germany., Petković H; Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia. hrvoje.petkovic@bf.uni-lj.si., Müller R; Department of Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)-Helmholtz Centre for Infection Research (HZI), and Department of Pharmacy, Saarland University Campus, Campus E8.1, 66123, Saarbrücken, Germany. rolf.mueller@helmholtz-hips.de.; German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, 38124, Braunschweig, Germany. rolf.mueller@helmholtz-hips.de.
المصدر: Microbial cell factories [Microb Cell Fact] 2020 Dec 19; Vol. 19 (1), pp. 230. Date of Electronic Publication: 2020 Dec 19.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: BioMed Central Country of Publication: England NLM ID: 101139812 Publication Model: Electronic Cited Medium: Internet ISSN: 1475-2859 (Electronic) Linking ISSN: 14752859 NLM ISO Abbreviation: Microb Cell Fact Subsets: MEDLINE
أسماء مطبوعة: Original Publication: London : BioMed Central, [2002-
مواضيع طبية MeSH: Genes, Bacterial* , Multigene Family*, Streptomyces/*genetics , Tetracyclines/*biosynthesis, Amycolatopsis/genetics ; Amycolatopsis/metabolism ; Anti-Bacterial Agents/biosynthesis ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Biosynthetic Pathways ; Cloning, Molecular ; Cosmids ; Metabolic Engineering ; Streptomyces/metabolism ; Tetracyclines/pharmacology
مستخلص: Background: Chelocardin (CHD) exhibits a broad-spectrum antibiotic activity and showed promising results in a small phase II clinical study conducted on patients with urinary tract infections. Importantly, CHD was shown to be active also against tetracycline-resistant Gram-negative pathogens, which is gaining even more importance in today's antibiotic crisis. We have demonstrated that modifications of CHD through genetic engineering of its producer, the actinomycete Amycolatopsis sulphurea, are not only possible but yielded even more potent antibiotics than CHD itself, like 2-carboxamido-2-deacetyl-chelocardin (CD-CHD), which is currently in preclinical evaluation. A. sulphurea is difficult to genetically manipulate and therefore manipulation of the chd biosynthetic gene cluster in a genetically amenable heterologous host would be of high importance for further drug-discovery efforts.
Results: We report heterologous expression of the CHD biosynthetic gene cluster in the model organism Streptomyces albus del14 strain. Unexpectedly, we found that the originally defined CHD gene cluster fails to provide all genes required for CHD formation, including an additional cyclase and two regulatory genes. Overexpression of the putative pathway-specific streptomyces antibiotic regulatory protein chdB in A. sulphurea resulted in an increase of both, CHD and CD-CHD production. Applying a metabolic-engineering approach, it was also possible to generate the potent CHD analogue, CD-CHD in S. albus. Finally, an additional yield increase was achieved in S. albus del14 by in-trans overexpression of the chdR exporter gene, which provides resistance to CHD and CDCHD.
Conclusions: We identified previously unknown genes in the CHD cluster, which were shown to be essential for chelocardin biosynthesis by expression of the full biosynthetic gene cluster in S. albus as heterologous host. When comparing to oxytetracycline biosynthesis, we observed that the CHD gene cluster contains additional enzymes not found in gene clusters encoding the biosynthesis of typical tetracyclines (such as oxytetracycline). This finding probably explains the different chemistries and modes of action, which make CHD/CD-CHD valuable lead structures for clinical candidates. Even though the CHD genes are derived from a rare actinomycete A. sulphurea, the yield of CHD in the heterologous host was very good. The corrected nucleotide sequence of the CHD gene cluster now contains all gene products required for the production of CHD in a genetically amenable heterologous host, thus opening new possibilities towards production of novel and potent tetracycline analogues with a new mode of action.
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معلومات مُعتمدة: J4-8226 Javna Agencija za Raziskovalno Dejavnost RS; P4-0116 Javna Agencija za Raziskovalno Dejavnost RS; P4-0116 Javna Agencija za Raziskovalno Dejavnost RS; TTU09.814 German Center for Infection Research (DZIF); TTU09.821 German Center for Infection Research (DZIF)
فهرسة مساهمة: Keywords: Actinobacteria; Antibiotics; Chelocardin; Heterologous expression; Natural product biosynthesis; Polyketide; Tetracyclines
المشرفين على المادة: 0 (Anti-Bacterial Agents)
0 (Bacterial Proteins)
0 (Tetracyclines)
N09QLW5PXU (chelocardin)
SCR Organism: Amycolatopsis sulphurea; Streptomyces albus
تواريخ الأحداث: Date Created: 20201220 Date Completed: 20210901 Latest Revision: 20210901
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC7749508
DOI: 10.1186/s12934-020-01495-x
PMID: 33341113
قاعدة البيانات: MEDLINE
الوصف
تدمد:1475-2859
DOI:10.1186/s12934-020-01495-x