Display Technologies for High-Throughput Enzyme Screening

CD Biosynsis offers specialized Enzyme Display Technology Services, serving as a core tool for High-Throughput Screening (HTS) to significantly accelerate directed evolution and enzyme discovery. Display technologies (such as Phage Display, Yeast Surface Display, mRNA Display, and Cell Surface Display) physically link the enzyme variant to its encoding gene, enabling highly efficient enrichment and sorting based on activity or binding affinity. These systems provide ultra-large library capacities, ranging from 10^8 to 10^13 variants, easily handling the vast mutant libraries generated by saturation mutagenesis or DNA shuffling. By integrating these display platforms with Fluorescence-Activated Cell Sorting (FACS), we can rapidly identify and recover rare enzyme variants with specific optimized properties, overcoming the limitations of traditional microplate screening and ensuring the precise evolution of activity, stability, or substrate specificity.

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Accelerating Directed Evolution with Ultra-High Capacity Systems

Traditional enzyme directed evolution faces two main challenges: the diversity of the mutant library and the throughput of the screening process. Display technologies solve both issues by establishing a physical "genotype-phenotype" link. In these systems, enzyme activity or binding affinity is converted into an easily detectable and sortable signal (typically fluorescence or magnetic bead binding). For example, Yeast Display combined with FACS allows us to sort the top 0.1% of the most active variants from millions of yeast cells within hours. mRNA Display achieves even higher library capacities, making it suitable for de novo design tasks that require exploring immense sequence space. We custom-tailor the most appropriate display platform based on the client's enzyme characteristics and project goals, ensuring optimal efficiency and accuracy in screening.

Core Enzyme Display Technology Platforms

Cellular/Viral Display Systems Eukaryotic Display Systems Cell-Free and Universal Display

High-Capacity Affinity Enrichment

Utilizing Phage and Cell Surface Display

Phage Display for Enzyme Engineering Service

Displaying enzyme variants on the surface of bacteriophage, allowing for selection and enrichment based on affinity binding to a target molecule (substrate or inhibitor). Library capacity up to 10^9.

Cell Surface Display for Enzyme Engineering Service

Anchoring enzyme variants onto the surface of bacterial (e.g., E coli) or other host cells, combined with FACS for sorting, suitable for screening under specific physiological conditions.

Ribosome Display

An in vitro cell-free translation system where the variant protein remains associated with its ribosomal complex, suitable for enzyme engineering with high stability requirements.

Function and Post-Translational Modifcations

Leveraging Yeast Surface Display

Yeast Surface Display for Enzyme Engineering Service

Utilizes the yeast cell surface display system, capable of eukaryotic folding and post-translational modifications, combined with FACS for 10^7 scale activity sorting.

FACS-Based Screening

Using high-precision Fluorescence-Activated Cell Sorting to sort single cells based on the intensity of the fluorescent signal generated by enzyme activity.

Off-Rate Selection

Controlling elution time in display technology to screen for enzyme variants with high affinity and slow dissociation rates from the substrate.

Maximum Exploration of Sequence Space

Leveraging mRNA Display

mRNA Display for Enzyme Engineering Service

An in vitro cell-free system where the enzyme variant is covalently linked to its mRNA template, offering the highest library capacity (up to 10^13) for de novo or large sequence exploration.

In Vitro Selection

All reactions are performed in a test tube, avoiding cell transformation limits and growth bias, suitable for cytotoxic or non-natural amino acid-modified enzymes.

Enzyme Activity Sorting

Development of enzyme activity reporter probes where the catalytic product is converted into a fluorescent signal for high-throughput activity enrichment cycles.

Enzyme Display Technology Screening Pipeline

Ultra-efficient enrichment from gene library to active variant.

Library Construction and Display

Target Binding and Selection

Enrichment and FACS Analysis

Recovery and Identification

Vector Preparation: Subcloning the mutant enzyme library into the specific display vector.

Expression and Display: Induce the host cells (e.g., yeast, bacteria, or phage) to express and display the enzyme variant on their surface.

Activity Labeling: Incubate the enzyme library with a fluorescently labeled substrate or target molecule, linking enzyme activity to a fluorescent signal.

Wash Enrichment: Remove low-affinity or low-activity variants via magnetic beads or multiple washing steps for preliminary enrichment.

FACS/Sorting: Subject the enriched cell suspension to Fluorescence-Activated Cell Sorting (FACS) for single-cell sorting.

Sequence Recovery: Extract DNA, perform PCR amplification on the sorted high-fluorescence cells, preparing for the next round of directed evolution.

Iterative Cycling: Repeat the above steps for multiple rounds of evolution to reach target performance goals.
Sequencing: Sequence the final successful clones to identify beneficial mutations.
Delivery: Optimized gene sequences and a detailed evolution report.

Unique Benefits of Display Technologies

Massive Throughput

mRNA Display offers capacity up to 10^13, covering all random and saturation mutant space.

Genotype-Phenotype Link

Mutant and its encoding gene are physically linked, ensuring accurate recovery of the corresponding DNA after selection.

Precise Quantitative Sorting

Combined with FACS, activity can be quantitatively graded based on fluorescent signal, enabling high-precision enrichment.

Suitable for Complex Enzymes

Yeast Display and similar systems support complex disulfide bond formation and post-translational modifications, suitable for eukaryotic enzymes.

Client Testimonials on Enzyme Display Technologies

"We successfully screened our 10^7 variant library using Yeast Surface Display combined with FACS, isolating an enzyme with a 15^\circC improvement in thermostability, far exceeding microplate screening efficiency."

Dr. Samuel Liu, R&D Director

"The Phage Display service helped us efficiently screen for enzymes with high binding affinity to a non-natural substrate, greatly simplifying the initial screening process."

Ms. Janet Chen, Lead Bioengineer

"For our de novo designed enzyme library, mRNA Display was the only option. It allowed us to explore 10^12 sequence space and successfully enrich for active clones."

Dr. Kenji Tanaka, Principal Scientist

FAQs about Enzyme Display Technologies Services

Still have questions?

Which display technique is best for screening ultra-large libraries?

If you need to explore library sizes up to 10^10 to 10^13 variants, mRNA Display for Enzyme Engineering Service is the optimal choice, as it is a cell-free system not limited by cell transformation efficiency.

If my enzyme requires post-translational modifications, which system should I choose?

If your enzyme requires complex folding, disulfide bond formation, or glycosylation, which are characteristic of eukaryotic post-translational modifications, we recommend using Yeast Surface Display for Enzyme Engineering Service.

Can display technologies screen for catalytic activity, not just binding affinity?

Yes, we screen for activity by developing "activity probes" or coupled reactions. For example, in Phage Display for Enzyme Engineering Service, the substrate can be covalently linked to the phage to screen for the ability to catalyze substrate conversion.

What is the advantage of Cell Surface Display over Yeast Display?

Cell Surface Display for Enzyme Engineering Service typically uses bacterial hosts, offering high transformation efficiency and faster growth. It is also convenient for screening under specific pH or temperature conditions.

How much does Metabolic Engineering services cost?

The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.

Do your engineered strains meet regulatory standards?

We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).

What to look for when selecting the best gene editing service?

We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.

Does gene editing allow customisability?

Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.

What is the process for keeping data private and confidential?

We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.