Tolypocladium inflatum Fermentation Services
As a leading CDMO, BOC Sciences has the ability to offer a broad range of microbial metabolites. We offer Tolypocladium inflatum fermentation services for the custom production of secondary metabolites such as cyclosporins. With our advanced technology, strong production capacity and comprehensive quality management system, we are able to meet the needs of our customers with high quality services and products at competitive prices.
Introduction of T. inflatum Fermentation
Tolypocladium inflatum, abbreviated as T. inflatum, is an ascomycete fungus that is commonly found in the soil. T. inflatum exists in both sexual and asexual states, the former being considered as an insect pathogen and the latter as a parthenogenic soil saprophyte. T. inflatum is capable of producing a variety of biologically active secondary metabolites and has been of interest to the pharmaceutical and biotechnology industries. It produces two important groups of metabolites, cyclosporin A and efrapeptin, both peptides with antifungal activity, which are produced via similar biosynthesis pathways. The gene clusters involved in biosynthesis in T. inflatum encode several enzymes, including non-ribosomal peptide synthase (NRPS) and a polyketide synthase (PKS).
Biosynthesis of Cyclosporin A in T. inflatum
Non-ribosomal peptide synthase (NRPS) contains modules responsible for sequential substrate uptake and chain elongation, including a core condensation-adenylation-thiolation domain, and an additional N-methylation domain. Substrates for NRPS include L-valine, L-leucine, L-alanine, glycine, D-alanine, 2-aminobutyric acid, etc. For the adenylation domain, NRPS generates an acyl adenylated amino acid, which is then covalently bound to phosphopantetheine via a thioester bond. Cyclization of linear peptide precursors is usually mediated by the terminal condensation domain of fungal NRPS. Given the antifungal activity of Cyclosporin A, comparing wild-type with Cyclosporin A-producing mutants of T. inflatum, cyclosporins production could benefit T. inflatum by conferring it a competitive edge over other fungi.
Our Services for T. inflatum Fermentation
We offer T. inflatum strain improvement and screening services. We analyze and identify genes involved in the biosynthesis of target secondary metabolites and apply techniques such as recombinant DNA and mutagenesis to the transformation of T. inflatum. Our goal is to screen for improved mutants of T. inflatum capable of overproducing the target secondary metabolites.
We offer controlled T. inflatum fermentation and fermentation scale-up services, from shake flask seed culture, seed tank culture to high volume fermenter culture. Fermentation process optimization includes medium optimization, which contains mainly carbon, nitrogen, amino acids and salts, as well as fermentation process selection, including submerged and solid state fermentation.
Secondary metabolites analysis
Secondary metabolites, such as cyclosporin A, are extracted from the fermentation broth of T. inflatum, and the titer of cyclosporin A is determined by HPLC. We provide detailed analytical reports of the target secondary metabolites.
T. inflatum Fermentation Conditions
- Aerobic conditions
- Autoclaved broth medium
- Medium in the pH range of 2 to 9
- Incubation temperature of 25-30 degrees C
- Stirring speed of 150 and 200 rpm
The Workflow of T. inflatum Fermentation
BOC Sciences' T. inflatum Fermentation
- Large supply of Cyclosporins, their analogues and related metabolites
- Obtained ISO 9001, ISO 22000 and ISO 14001 certifications
- Complete quality management system
- Different sizes of fermenters from 30 L to 30 KL
- Production process control and tracking
- Patented and proprietary manufacturing methods
