Protein Expression and Purification

Saccharomyces cerevisiae Protein Expression and Purification Services

Reliable recombinant protein production in S. cerevisiae with scalable fermentation and multi-step purification.

Scalable expression from shake flask to bioreactor (up to 20 L as scoped)
Multi-step purification: affinity, ion exchange.
Comprehensive QC: SDS-PAGE, western blot, endotoxin assay.

We tailor expression conditions (promoter, strain, induction) and purification protocols to your target protein's solubility, activity, and final purity requirements, ensuring project-specific success. Our dedicated project team manages every step from gene synthesis through final QC delivery, with regular milestone updates and transparent reporting throughout the engagement.

Gene synthesis and codon optimization for S. cerevisiae expression
Small-scale expression screening and optimization
Fermentation scale-up (flask or bioreactor)

Strain & Vector Optimization

Fermentation Scale-Up (up to 20 L)

QC & Documentation Package

Our Approach

Codon Optimization & Gene Synthesis

Optimization for S. cerevisiae codon usage to evaluate expression levels. All constructs are sequence-verified before transformation.

Fermentation Optimization

Shake flask and bioreactor scale-up with controlled pH, temperature, and induction timing to evaluate volumetric yield.

QC & Documentation

Routine purity (SDS-PAGE), identity (western blot), endotoxin, and concentration assays. Full data package delivered with the protein.

Deliverables

What You Receive

Standard deliverables are scoped per project; additional documentation or custom QC can be added upon request.

Purified Protein

Purified target protein in agreed buffer (liquid or lyophilized)
Concentration determined by A280 or BCA assay
Purity ≥ project-dependent performance that is verified during development by SDS-PAGE (target purity defined per project)
Endotoxin level <1 EU/mg (when scoped)

Documentation

Expression and purification report (methods, conditions, chromatograms)
QC data package (SDS-PAGE gel image, western blot if applicable, endotoxin results)
Sequence-verified expression vector map and sequence file
Glycerol stock of production strain

Service Scope

Customizable Parameters

Each project is designed around your target protein's specific requirements. Typical parameters considered include:

Parameter	Typical project scope
Host Strain	Saccharomyces cerevisiae (multiple strains available; selection based on glycosylation, protease profile, and growth characteristics)
Expression Vector / Promoter	Inducible (GAL1, CUP1, ADH2) or constitutive (PGK1, TEF1, GPD) as required for optimal expression
Expression Scale	Shake flask (scale scoped per project–scale scoped per project) or bioreactor (scale scoped per project) per project; larger scales as scoped
Purification Methods	Affinity (His-tag, GST-tag, MBP-tag), ion exchange (IEX), size exclusion chromatography (SEC)
QC Assays	SDS-PAGE, western blot (if antibody provided), endotoxin (LAL), concentration (A280/BCA)
Deliverable Form & Buffer	Liquid concentrate or lyophilized in customer-specified buffer; final formulation agreed during project design
Project Timeline	Scoped per project after consultation
Technical Support	Named scientific contact at project start; milestone review calls; email response within 1 business day

Workflow

Project Execution

Our standard five-stage process ensures structured development, regular reporting, and transparent communication.

Gene Design & Vector Construction

We synthesize codon-optimized cDNA for S. cerevisiae and clone into your chosen expression vector. The construct is sequence-verified before proceeding to transformation.

Transformation & Expression Screening

The vector is transformed into the selected S. cerevisiae strain. Small-scale cultures are screened for expression level, solubility, and induction conditions using SDS-PAGE and western blot.

Fermentation Scale-Up

The best-performing clone is scaled up from shake flask to a 1–20 L bioreactor, with controlled parameters (pH, temperature, dissolved oxygen) to evaluate biomass and protein yield.

Cell Lysis & Protein Extraction

Cells are harvested, lysed (bead beating or high-pressure homogenizer), and clarified by centrifugation. Soluble and insoluble fractions are analyzed to guide purification strategy.

Purification & QC

Target protein is captured by affinity chromatography, polished by IEX or SEC, and concentrated. Final QC includes SDS-PAGE, western blot, endotoxin assay, and concentration measurement.

Applications

Enzyme Production

Recombinant enzymes for biocatalysis, industrial processing, or metabolic pathway reconstitution. S. cerevisiae offers proper folding and post-translational modifications for many eukaryotic enzymes.

High-yield expression with codon optimization
Activity-preserved purification (buffers tailored to enzyme)
Scale-up to bioreactor for gram-scale yields (as scoped)

Therapeutic Protein Research

Production of vaccine antigens, cytokines, or antibody fragments for preclinical studies. S. cerevisiae provides a low-cost, scalable platform with a eukaryotic secretion pathway. Note: any subsequent in vivo or animal studies requiring the protein are conducted under the customer's own IACUC-approved protocols.

Secretion or intracellular expression as desired
Endotoxin-controlled purification (≤1 EU/mg when required)
Documentation supporting early-stage development

Structural Biology

Proteins for X-ray crystallography, NMR spectroscopy, or cryo-EM. High purity and homogeneity are essential, along with isotope labeling options for structural analysis.

Final purity ≥ project-dependent performance that is verified during development by SEC and SDS-PAGE (scoped per target)
Optional isotopic enrichment (15N, 13C) for NMR studies
Size exclusion chromatography as final polishing step

Frequently Asked Questions

Common Inquiries

Answers to typical questions about our S. cerevisiae expression and purification service.

Projects generally require 6 to 10 weeks from sequence confirmation to delivery of purified protein. This includes gene synthesis, cloning, transformation, expression screening, fermentation scale-up, purification development, and QC. The exact timeline depends on target protein complexity, required yield, and any iterative optimization steps.
Our standard service focuses on soluble protein expression and purification. Membrane protein production requires specialized detergents, solubilization protocols, and often different host strain choices. If your target is a membrane protein, please contact us with details so we can assess feasibility and discuss a custom scope.
Purity targets are agreed at project initiation.Typical affinity purification yields ≥ project-dependent performance that is verified during development by SDS-PAGE.
Protein is delivered in your specified buffer, either as a liquid concentrate (shipped on dry ice) or lyophilized (shipped at ambient). You receive a full data package: expression and purification report, SDS-PAGE gel image, western blot (if antibody was provided), endotoxin assay results, and concentration measurement. Additional certificates of analysis can be provided upon request.

Scientific Literature

References

Wang G, Møller-Hansen I, Babaei M, et al. Transportome-wide engineering of Saccharomyces cerevisiae. Metabolic engineering. 2021;64:52-63. View on PubMed
Yip MCJ, Bodnar NO, Rapoport TA. Ddi1 is a ubiquitin-dependent protease. Proceedings of the National Academy of Sciences of the United States of America. 2020;117(14):7776-7781. View on PubMed
Long KS, Cedervall T, Walch-Solimena C, et al. Phosphorylation of the Saccharomyces cerevisiae La protein does not appear to be required for its functions in tRNA maturation and nascent RNA stabilization. RNA (New York, N.Y.). 2001;7(11):1589-602. View on PubMed
Haslem L, Brown M, Zhang XA, et al. Purification of Membrane Proteins Overexpressed in Saccharomyces cerevisiae. Methods in molecular biology (Clifton, N.J.). 2022;2507:143-173. View on PubMed
Nozaki LY, Bulka NR, Dos Reis KL, et al. Expression of the Fusarium graminearum galactose oxidase GaoA in Saccharomyces cerevisiae. Protein expression and purification. 2025;227:106637. View on PubMed

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