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

Content Provider	Springer Nature : BioMed Central
Author	Lukežič, Tadeja Pikl, Špela Zaburannyi, Nestor Remškar, Maja Petković, Hrvoje Müller, Rolf
Abstract	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.
Related Links	https://microbialcellfactories.biomedcentral.com/counter/pdf/10.1186/s12934-020-01495-x.pdf
Ending Page	13
Page Count	13
Starting Page	1
File Format	HTM / HTML
ISSN	14752859
DOI	10.1186/s12934-020-01495-x
Journal	Microbial Cell Factories
Issue Number	1
Volume Number	19
Language	English
Publisher	BioMed Central
Publisher Date	2020-12-19
Access Restriction	Open
Subject Keyword	Applied Microbiology Biotechnology Microbiology Microbial Genetics and Genomics Enzymology Genetic Engineering Antibiotics Polyketide Heterologous expression Actinobacteria Natural product biosynthesis Tetracyclines Chelocardin
Content Type	Text
Resource Type	Article
Subject	Applied Microbiology and Biotechnology Bioengineering Biotechnology
Journal Impact Factor	4.3/2023
5-Year Journal Impact Factor	5.5/2023

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Uncovering the genetic basis of antiviral polyketide limocrocin biosynthesis through heterologous expression

Biosynthesis of C-nucleoside antibiotics in actinobacteria: recent advances and future developments

A Streptomyces coelicolor host for the heterologous expression of Type III polyketide synthase genes

Engineered fungal polyketide biosynthesis in Pichia pastoris: a potential excellent host for polyketide production

Successful biosynthesis of natural antioxidant ergothioneine in Saccharomyces cerevisiae required only two genes from Grifola frondosa

Engineering of Streptomyces lividans for heterologous expression of secondary metabolite gene clusters

Genome-wide screen identifies new set of genes for improved heterologous laccase expression in Saccharomyces cerevisiae

Synthesis of acridone derivatives via heterologous expression of a plant type III polyketide synthase in Escherichia coli

Exploring the biocombinatorial potential of benzoxazoles: generation of novel caboxamycin derivatives

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

Similar Documents

Uncovering the genetic basis of antiviral polyketide limocrocin biosynthesis through heterologous expression

Biosynthesis of C-nucleoside antibiotics in actinobacteria: recent advances and future developments

A Streptomyces coelicolor host for the heterologous expression of Type III polyketide synthase genes

Engineered fungal polyketide biosynthesis in Pichia pastoris: a potential excellent host for polyketide production

Successful biosynthesis of natural antioxidant ergothioneine in Saccharomyces cerevisiae required only two genes from Grifola frondosa

Engineering of Streptomyces lividans for heterologous expression of secondary metabolite gene clusters

Genome-wide screen identifies new set of genes for improved heterologous laccase expression in Saccharomyces cerevisiae

Synthesis of acridone derivatives via heterologous expression of a plant type III polyketide synthase in Escherichia coli

Exploring the biocombinatorial potential of benzoxazoles: generation of novel caboxamycin derivatives

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