Methanol-free Komagataella phaffii (Pichia pastoris) system for scalable, cost-effective industrial protein production

BRAIN Biotech has developed a proprietary, patent-pending, methanol-free and inducible production system for Komagataella phaffii (Pichia pastoris), offering a safe, scalable, and cost-effective solution. This next-generation production system enables high-yield protein production without the hazards or regulatory hurdles typically associated with methanol-inducible systems. 

Why was it necessary to development a methanol-free system?

The methylotrophic yeast K. phaffii has long been a cornerstone of recombinant protein production, valued for its ability to achieve high cell densities, tightly regulate gene expression via methanol-inducible promoters, and perform eukaryotic post-translational modifications. However, the use of methanol introduces significant technical and regulatory challenges, including:

• Safety risks & high cost: Methanol is volatile, flammable, and toxic. Its use requires explosion-proof facilities, continuous monitoring, and specialized handling - adding complexity and cost.
• High oxygen demand & heat production: Methanol metabolism demands intense aeration and cooling, escalating operational costs and limiting cell density.
• Cellular stress: Methanol metabolism leads to reactive oxygen species (ROS) formation, triggering stress responses that impair protein production and reduce cell viability.
• Scale-up limitations: Only a limited number of CMOs are equipped to handle the high oxygen and cooling demands of methanol-based processes at commercial scale, creating bottlenecks for industrialization.

BRAIN Biotech's methanol-free K. phaffii production platform

Our innovative expression system utilizes an inducible, methanol-free promoter, eliminating the need for hazardous substances. The employed inducers are non-toxic, food-grade, cheap and widely available, simplifying supply chains and reducing regulatory risk.

The inducible promoter allows:

• temporal control of expression
• reduction of cellular stress during the growth phase
• enabling a more robust and predictable production phase

The result is a platform capable of delivering yields suitable for industrial applications, without compromising on safety, sustainability, or regulatory compliance.

How can BRAIN Biotech's customers benefit from this?

Through our contract research services, our customers gain access to the methanol-free system and, if desired, can additionally make use of our end-to-end services to reduce time-to-market for recombinant protein products. We provide a fully integrated manufacturing solution, combining cutting-edge molecular biology with manufacturing and regulatory expertise.

• CRISPR-based strain engineering: Utilize our proprietary BMC genome editing platform for precise gene knockouts, metabolic rewiring, and co-expression of chaperones or secretion enhancers.
   
• Integrated strain & process optimization: Strain design and fermentation optimization are carried out in parallel for synergistic yield improvements. Expertise in downstream processing enables rapid development of purification strategies at lab scale.
   
• Industrial scale-up & manufacturing: Process scale-up to 10 m³ is available at our production site at Biocatalysts Ltd. Facilities are food-grade, kosher, and halal-certified, ensuring compliance for global markets.
   
• Compliance & regulatory support: All strains are constructed without the use of antimicrobial resistance markers (AMR-free), aligning with food and pharma safety standards. We provide guidance on dossier preparation, safety assessments, and EFSA or equivalent submissions. Support across the full regulatory lifecycle, from early development through to market entry.

BRAIN Biotech’s methanol-free K. phaffii platform represents a next-generation solution for recombinant protein production - combining easier utilization, safety, scalability, and regulatory readiness - for a safer, faster and scalable protein production.