[P001] Poster

12:10 PM - 1:00 PM JST(3:10 AM - 4:00 AM UTC)

[P01-071] Process development and strain engineering for pneumocandin B₀ enhancement from Glarea lozoyensis ATCC 20868

○Anshu Baldia¹, Kashyap Kumar Dubey¹ (1. School of Biotechnology, Jawaharlal Nehru university, New Delhi (India))

Keywords:

Pneumocandin B0,Antifungal agent,Strain engineering,Statistical technique

Pneumocandin B₀, a lipohexapeptide produced by the filamentous fungus Glarea lozoyensis, serves as the key precursor for caspofungin synthesis. Therefore, improving pneumocandin B₀ production is crucial for enhancing industrial-scale antifungal drug manufacturing. This study focuses on the development of an optimized bioprocess and strain improvement for pneumocandin B₀ production. Initially, carbon and nitrogen sources were optimized using a one-variable-at-a-time approach. Soybean meal and mannitol were identified as the optimal nitrogen and carbon sources, respectively. Growth conditions were further optimized using response surface methodology with central composite design (RSM-CCD). Key parameters, including carbon sources (mannitol and fructose), nitrogen source (soybean meal), and pH, were evaluated. Under optimized conditions, pneumocandin B₀ production increased significantly, resulting in a 2.3-fold enhancement. To further improve production, strain engineering was performed using both physical and chemical mutagenesis. Gamma irradiation was applied for physical mutagenesis, with approximately 90% mortality observed at 250 Gy, from which stable mutants were selected. The best-performing gamma mutant exhibited a 1.74-fold increase in pneumocandin B₀ production compared with the wild-type strain. Chemical mutagenesis using methyl methanesulfonate (MMS) was also employed, with optimal treatment conditions of 0.2% MMS for 60 min, resulting in 87% mortality. The top-performing MMS mutant (M7) demonstrated a 2.2-fold increase in production relative to the wild type. Morphological and microscopic analyses revealed significant structural changes in mutant strains, including altered hyphal architecture and increased fragmentation, which were likely associated with enhanced metabolite secretion. Overall, this study presents an integrated strategy combining media optimization and mutagenesis to significantly enhance pneumocandin B₀ production and provides a strong foundation for developing efficient and scalable industrial processes for caspofungin manufacture. In conclusion, this study presents an integrated strategy combining media optimization, mutagenesis, and scale-up validation to significantly enhance pneumocandin B₀ production. The findings provide a strong foundation for developing efficient and scalable industrial processes for caspofungin production, contributing to improved availability of life-saving antifungal therapies.