ArticleApplied microbiology and biotechnology2016
Production of the polyketide 6-deoxyerythronolide B in the heterologous host Bacillus subtilis.
Article in Applied microbiology and biotechnology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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18 citing papers in PubMed, 37 citations in OpenAlex.
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- A host-vector toolbox for improved secretory protein overproduction in Bacillus subtilis.Applied microbiology and biotechnology · 2022Article
- Efficient Large-Scale and Scarless Genome Engineering Enables the Construction and Screening ofInternational journal of molecular sciences · 2022Article
- Heterologous Production of 6-Deoxyerythronolide B inMetabolites · 2020Article
- Article
- A metagenomic strategy for harnessing the chemical repertoire of the human microbiome.Science (New York, N.Y.) · 2019Article
- Genetic platforms for heterologous expression of microbial natural products.Natural product reports · 2019Review
- Leveraging synthetic biology for producing bioactive polyketides and non-ribosomal peptides in bacterial heterologous hosts.MedChemComm · 2019Review
- Recent advances in CRISPR/Cas9 mediated genome editing in Bacillus subtilis.World journal of microbiology & biotechnology · 2018Review
- Acyltransferases as Tools for Polyketide Synthase Engineering.Antibiotics (Basel, Switzerland) · 2018Review
- Omics based approach for biodiscovery of microbial natural products in antibiotic resistance era.Journal, genetic engineering & biotechnology · 2018Review
- Engineered polyketides: Synergy between protein and host level engineering.Synthetic and systems biotechnology · 2017Review
- A Highly Efficient CRISPR-Cas9-Mediated Large Genomic Deletion inFrontiers in microbiology · 2017Article
- Simple and rapid direct cloning and heterologous expression of natural product biosynthetic gene cluster in Bacillus subtilis via Red/ET recombineering.Scientific reports · 2016Article
- The fed-batch principle for the molecular biology lab: controlled nutrient diets in ready-made media improve production of recombinant proteins in Escherichia coli.Microbial cell factories · 2016Review
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Polyketides, such as erythromycin, are complex natural products with diverse therapeutic applications. They are synthesized by multi-modular megaenzymes, so-called polyketide synthases (PKSs). The macrolide core of erythromycin, 6-deoxyerythronolide B (6dEB), is produced by the deoxyerythronolide B synthase (DEBS) that consists of three proteins each with a size of 330-370 kDa. We cloned and investigated the expression of the corresponding gene cluster from Saccharopolyspora erythraea, which comprises more than 30 kb, in Bacillus subtilis. It is shown that the DEBS genes are functionally expressed in B. subtilis when the native eryAI-III operon was separated into three individual expression cassettes with optimized ribosomal binding sites. A synthesis of 6dEB could be detected by using the acetoin-inducible acoA promoter and a fed-batch simulating EnBase-cultivation strategy. B. subtilis was capable of the secretion of 6dEB into the medium. In order to improve the 6dEB production, several genomic modifications of this production strain were tested. This included the knockout of the native secondary metabolite clusters of B. subtilis for the synthesis of surfactin (26 kb), bacillaene (76 kb), and plipastatin (38 kb). It is revealed that the deletion of the prpBD operon, responsible for propionyl-CoA utilization, resulted in a significant increase of the 6dEB product yield when exogenous propionate is provided. Although the presented B. subtilis 6dEB production strain is not competitive with established Escherichia coli 6dEB production strains, the results of this study indicate that B. subtilis is a suitable heterologous host for the secretory production of a complex polyketide.
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