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Enzyme Yeast Display Service

Tek Biotech is committed to providing high-quality enzyme yeast display service to our customers to support their downstream research and development work, such as enzyme activity validation, functional enzyme screening (flow-through screening), development of high-activity enzymes, and in vitro directed evolution of proteases. Tek Biotech has many years of experience in enzyme yeast display service, and is able to provide customers with a variety of customized enzyme yeast display service, including, but not limited to, single enzyme display, trimer codonmutant enzyme library display, NNK mutant enzyme library display, error-prone PCR mutant enzyme library display, and many other enzyme yeast display services.

Yeast display technology is the fusion expression of enzyme sequences/enzyme mutation library sequences with the lectin Aga2p, where the Aga2p protein subunit binds to the Aga1p protein subunit immobilized on the yeast cell wall via two disulfide bonds (i.e., the carrier protein anchors the protease (with a specific tag) to the outer surface of the yeast cell in an active form) .The yeast strains induced to express the target enzyme proteins are screened in combination with flow sorting technology. Yeast display technology makes the target protease more stable by immobilizing it on the yeast surface, and the protease is close to its natural spatial conformation, which can be verified by ELISA, flow assay and other methods. Based on the yeast display technology, Teckbio can provide customers with high quality enzyme mutation libraries for yeast display, and the library capacity of yeast libraries can be up to 10^7-10^8, and the diversity of libraries, insertion rate and positive rate can be more than 90%, which can satisfy the quality requirements of all kinds of customers for enzyme and yeast display libraries. Meanwhile, Teckbio can also provide one-stop technical services such as downstream flow validation, ELISA validation, flow sorting, etc. Customers only need to provide specific enzyme sequence information and the sites to be mutated (continuous mutation or discontinuous point mutation, etc.), and Teckbio's scientists will be able to carry out a reasonable program design and personalized customization according to the needs of the customers, which will help customers to develop high activity enzymes for their scientific research and development projects. Teckbio's scientists are able to design and customize the program according to the customer's needs, which will help the customer's research projects and the development of highly active enzymes.

█ Enzyme Yeast Display Library Construction Service

Tek Biotech has established a complete yeast demonstration system, which can provide a variety of high-quality customized yeast demonstration services, including but not limited to VHH antibody yeast demonstration, VHH antibody CDR region mutation library yeast demonstration, scFv antibody yeast demonstration, protein yeast demonstration, enzyme yeast demonstration, enzyme mutation library yeast demonstration and so on, which can meet the scientific research needs of different customers. The service flow of enzyme gene random mutation library yeast display is shown in Figure 1.

Enzyme Yeast Display Service-Tekbiotech.jpg

Figure 1 Yeast Display Service Process for Enzyme Gene Random Mutation Library

█ Content and Periodicity of Services

Step

Service Content

Period

Individual enzyme yeast display

(1) Protease gene synthesis + PCR amplification of target gene;

(2) Vector construction and transformation: protease gene splicing of yeast display vector, electroshock transformation of yeast strain;

(3) Strain PCR validation;

(4) Delivery: display yeast strains as well as induction cultures, standard lab reports

4-5 weeks

Enzyme mutation library yeast display

(1) Construct a gene pool using trimer codon/NNK/error-prone PCR.

(2) Construct yeast vector gene pool

(3) Vector construction and transformation: electroshock transformation of yeast strain;

(4) Randomly pick single clones for strain PCR verification + library NGS sequencing verification;

(5) Delivery: 2-3 ml of library, library volume >10^8

4-6 weeks

█ Service Advantages

-- Multiple random mutagenesis options: trimer codon, NNK, error-prone PCR, etc.

-- Multiple enzyme display options: individual enzyme proteins can be displayed, and enzyme mutation libraries can be displayed (including but not limited to trimer codon mutation libraries, NNK mutation libraries, error-prone PCR mutation libraries).

-- Proven technology for enzyme yeast display: enzyme mutation libraries with library sizes up to 10^7-10^8, with insertion rates >90%.

-- High standard of delivery: library quality assurance to support customers' downstream experiments.

-- Traceability of experimental records: QC standards for libraries, Chinese and English experimental reports, original experimental records.

-- Provide a series of downstream experimental services, such as flow validation, library screening and so on.

-- One-to-one customized solutions, designing the best constructs (e.g. mutant libraries, labels, vectors, validation protocols, etc.) according to customers' needs, to meet the needs of all kinds of customers' scientific research projects.

-- One-stop service: one-stop technical service from program design, gene/library synthesis, vector construction, strain transformation, induced expression, flow-through validation to library screening.

Enzyme Yeast Display Service Frequently Asked Questions

Q: Enzymes may not be correctly located at the target site of yeast cells, affecting their functional expression.

A: Optimize the signal peptide sequence to ensure that the enzyme can be correctly located on organelles or cell membranes to achieve its functional expression.
Q: Enzymes may not fold or assemble correctly within yeast cells, leading to structural abnormalities.

A: Select appropriate signal peptides and expression hosts, optimize culture conditions to promote correct folding and assembly of enzymes; Using molecular screening techniques or protein engineering methods to modify enzymes and improve their stability and activity.
Q: The expression level of enzymes in yeast is low, which affects enzyme activity and yield.

A: Optimize promoter and signal peptide sequences, adjust culture conditions (such as temperature, pH, medium composition, etc.), select appropriate yeast strains and expression vectors to improve enzyme expression levels and activity.

Q: Enzymes may not be correctly located at the target site of yeast cells, affecting their functional expression.

A: Optimize the signal peptide sequence to ensure that the enzyme can be correctly located on organelles or cell membranes to achieve its functional expression.
Q: Enzymes may not fold or assemble correctly within yeast cells, leading to structural abnormalities.

A: Select appropriate signal peptides and expression hosts, optimize culture conditions to promote correct folding and assembly of enzymes; Using molecular screening techniques or protein engineering methods to modify enzymes and improve their stability and activity.
Q: The expression level of enzymes in yeast is low, which affects enzyme activity and yield.

A: Optimize promoter and signal peptide sequences, adjust culture conditions (such as temperature, pH, medium composition, etc.), select appropriate yeast strains and expression vectors to improve enzyme expression levels and activity.

Consult Now Enzyme Yeast Display Service

Yeast Display Platform

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Enzyme Yeast Display Service Frequently Asked Questions

Q: Enzymes may not be correctly located at the target site of yeast cells, affecting their functional expression.

Q: Enzymes may not fold or assemble correctly within yeast cells, leading to structural abnormalities.

Q: The expression level of enzymes in yeast is low, which affects enzyme activity and yield.

Q: Enzymes may not be correctly located at the target site of yeast cells, affecting their functional expression.

Q: Enzymes may not fold or assemble correctly within yeast cells, leading to structural abnormalities.

Q: The expression level of enzymes in yeast is low, which affects enzyme activity and yield.

Consult Now Enzyme Yeast Display Service