Yeast Protein Expression Service | Pichia pastoris & More

Yeast Protein Expression Service

High-yield · Scalable · Eukaryotic Modifications
Pichia pastoris expression platform with codon optimization and high-density fermentation – from gene to purified protein.
Common Strains
GS115

Mut⁺ (methanol utilization fast), AOX1 high-expression, ideal for most recombinant proteins.

KM71

Mutˢ slow methanol utilization, lower methanol consumption, suitable for toxic proteins.

X33

Wild-type Zeocin resistance, high-copy integration, stable yields.

SMD1168

Protease-deficient strain, reduces product degradation, improves integrity.

Optimal strain recommendation based on protein characteristics & codon optimization available.
System Advantages
Low-cost media, simple components
Fast growth (doubling time ~90 min)
Eukaryotic folding: disulfide bonds, glycosylation, phosphorylation
Secretion system for easier downstream purification
Scalable to >500 L industrial scale
Genetic stability: chromosomal integration, high-copy maintenance
Methanol-inducible AOX1 promoter (tight regulation)
Low mannosylation pattern – reduced immunogenicity
Service Process
StepContentLead Time
Step 1Gene synthesis & codon optimization (for Pichia pastoris)2–4 weeks
Step 2Vector construction & subcloning (pPICZα, pPIC9K etc.)1 week
Step 3Transformation & positive clone screening (high-copy/antibiotic gradient)2–3 weeks
Step 4Small-scale expression (shake flask, SDS-PAGE/WB verification)1–2 weeks
Step 5High-density fermentation & purification (1L–100L optional)2–4 weeks
Total7–10 weeks (including expression, purification & QC)
Technical Flowchart
1
Gene Optimization
Codon optimization + synthesis
2
Vector Construction
Subcloning → linearization
3
Electroporation
Competent yeast + homologous recombination
4
Screening
High-copy clone / phenotype selection
5
Shake-flask Test
Induction & expression validation
6
Fermentation & Purification
High-density culture / multi-step chromatography
Quality controls: SDS-PAGE, WB, endotoxin/activity assays (optional)
Expression Level Reference

Shake Flask

200 ml – 1 L scale
mg range (1–50 mg/L, protein-dependent)

Bioreactor / Fermenter

5 L – 100 L scale
hundreds mg to grams (typical 0.1–2 g/L)

Industrial Fermentation

500 L and above
gram+ level for pre-clinical/industrial supply

* Expression yields vary significantly depending on protein nature, molecular weight and folding difficulty. Contact our team for feasibility evaluation.
Protein Type Suitability

Well suited

  • Secreted proteins / soluble intracellular proteins
  • Proteins requiring disulfide bonds & glycosylation
  • Enzymes, cytokines, growth factors
  • Antibody fragments (scFv, Fab)
  • Vaccine antigens & industrial enzymes

More challenging (custom strategy needed)

  • Very large proteins (>150 kDa)
  • Complex human-like glycosylation (glyco-engineered strains available)
  • Multi-pass transmembrane proteins (membrane protein optimization)
  • Host-toxic proteins (special induction / conditional expression)
Representative Cases

Cytokine Interleukin analogue (~18 kDa) secreted in Pichia pastoris; fermenter yield >200 mg/L, single-step affinity purity >95%, active confirmed by ELISA.

Single-chain antibody (scFv) Tumor-targeting scFv: shake-flask screening ~12 mg/L; after optimization 5L fermenter reached 110 mg/L, recovery 80%.

Industrial enzyme Thermostable cellulase (~55 kDa) at 500 L scale; wet cell weight >300 g/L; purified specific activity 420 U/mg – suitable for industrial applications.

Above data represent typical project outcomes; actual yields depend on specific sequences & structure. Full QC from cloning to final protein provided.

Frequently Asked Questions
Does the yeast system ensure correct protein folding?
Pichia pastoris possesses ER and Golgi apparatus, facilitating disulfide bond formation and partial glycosylation. Many complex proteins are correctly folded. We also offer co-expression of chaperones or fusion tags for improved folding.
Are tags present in secreted proteins? Can they be removed?
Common tags include His, Myc, HA; tag-free expression is also available. For tag removal we provide protease cleavage strategies (TEV, EK).
Does yeast glycosylation affect protein activity compared to mammalian systems?
High mannose pattern may increase immunogenicity for certain therapeutics, but it is acceptable for diagnostic reagents, industrial enzymes and many research applications. Humanized glycosylation variants are available upon request (glyco-engineered strains).
Can you express large proteins (>100 kDa)?
Yes, we have successfully expressed kinase fusions up to ~120 kDa, though yields may decrease. A pilot-scale feasibility test is recommended.
Request a Quote
We will respond within 24h with a preliminary proposal and service details.
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