Engineering of Fungal Chassis for Biosynthesis

Chassis selection depends on your target molecule and production requirements:

• S. cerevisiae: Best for terpenoids, complex eukaryotic proteins, and compounds requiring P450 enzymes. GRAS status and well-established tools.
• P. pastoris: Ideal for secreted proteins requiring high yields. Superior protein secretion machinery and high cell density fermentation capability.
• A. niger: Industry standard for industrial enzymes and organic acids. Exceptional secretion capacity and low pH tolerance.
• Y. lipolytica: Excellent for lipid-derived products and compounds requiring acetyl-CoA overflow metabolism.

Our team will provide detailed recommendations based on your target molecule, titer requirements, and scale-up plans during the initial consultation.

Project timelines vary based on complexity and scope:

• S. cerevisiae: 8-12 weeks for initial strain construction with one DBTL cycle
• P. pastoris: 10-14 weeks due to additional clone screening for secretion efficiency
• A. niger: 12-16 weeks as transformation efficiency requires more screening

Complex multi-gene pathways or iterative evolution campaigns will extend timelines accordingly. Express options are available for time-sensitive projects.

We employ multiple genome editing approaches:

• CRISPR-Cas9 RNP delivery: Our preferred method for marker-free, scarless editing with >90% efficiency in S. cerevisiae
• Cas12a/Cpf1: Used for multiplexed gene editing when multiple targets need simultaneous modification
• dCas9 transcriptional activation/repression: For dynamic pathway regulation without permanent genomic changes
• Traditional homologous recombination: Still valuable for large insertions or specific applications

Pathway balancing is critical for optimal production. Our approach includes:

• Promoter library screening: Characterized promoter collections with 100-1000x expression range
• RBS engineering: Translational level fine-tuning using characterized RBS variants
• Cofactor engineering: NADPH/NADH balance optimization for oxidative pathways
• Transporter identification: Adding exporter genes to relieve product toxicity
• Multi-omics guided iteration: Fluxomic analysis to identify and relieve bottlenecks

Yes, we offer comprehensive scale-up support:

• Shake flask to benchtop: Standard included deliverable with each strain
• 5-50L bioreactor optimization: Fed-batch strategy development and process parameters
• Industrial scale-up: Collaboration with manufacturing partners for commercial production
• Technical transfer: Complete documentation package for technology transfer to CMOs

Scale-up add-ons are available separately or as part of comprehensive development programs.

Every delivered strain undergoes comprehensive QC:

• Whole genome sequencing: Verification of all genetic modifications and absence of off-target effects
• Metabolite analysis: GC-MS/LC-MS quantification of target product and byproducts
• Stability testing: Continuous cultivation without selective pressure to confirm genomic stability
• Growth characterization: Growth curve comparison with parental strain
• Documentation: Complete strain construction records, plasmid maps, and fermentation protocols

Absolutely. We routinely handle:

• Client-provided gene sequences: We can clone and express any gene of interest
• Uncharacterized enzymes: Functional expression and characterization services available
• Novel biosynthetic pathways: Heterologous reconstitution in fungal chassis
• IP considerations: We can work under strict confidentiality and accommodate specific IP requirements

NDA agreements are standard practice for all projects involving proprietary materials.

Titers depend heavily on the specific compound, pathway complexity, and metabolic burden. Representative examples:

• Terpenoids (farnesene): 10-130+ g/L in optimized strains
• Phenolic acids (caffeic acid): 5-10 g/L achievable
• Organic acids (itaconic acid): 5-9 g/L in engineered A. niger
• Secreted proteins: 1-25+ g/L depending on protein characteristics

Initial titers from first-generation strains typically range from mg/L to low g/L. DBTL cycles iteratively improve production.