ArticleScientific reports2016
In vitro flow cytometry-based screening platform for cellulase engineering.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 59 citations in OpenAlex.
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- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Flow Cytometry: The Next Revolution.Cells · 2023Review
- High-throughput screening of microbial strains in large-scale microfluidic droplets.Frontiers in bioengineering and biotechnology · 2023Article
- Role of metagenomics in prospecting novel endoglucanases, accentuating functional metagenomics approach in second-generation biofuel production: a review.Biomass conversion and biorefinery · 2023Review
- Multi-color flow cytometer with PIC-based structured illumination.Biomedical optics express · 2022Article
- Microfluidic screening and genomic mutation identification for enhancing cellulase production in Pichia pastoris.Biotechnology for biofuels and bioproducts · 2022Article
- High-throughput screening, next generation sequencing and machine learning: advanced methods in enzyme engineering.Chemical communications (Cambridge, England) · 2022Review
- Fungal cellulases: protein engineering and post-translational modifications.Applied microbiology and biotechnology · 2022Review
- SpeedyGenesXL: an Automated, High-Throughput Platform for the Preparation of Bespoke Ultralarge Variant Libraries for Directed Evolution.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey.Angewandte Chemie (International ed. in English) · 2021Review
- Droplet-based microfluidic platform for high-throughput screening of Streptomyces.Communications biology · 2021Article
- Consolidated Bioprocessing: Synthetic Biology Routes to Fuels and Fine Chemicals.Microorganisms · 2021Review
- A High-Throughput Screening System Based on Fluorescence-Activated Cell Sorting for the Directed Evolution of Chitinase A.International journal of molecular sciences · 2021Article
- Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites.Frontiers in plant science · 2021Article
- Cell-free protein synthesis andRSC advances · 2020Article
- Split & mix assembly of DNA libraries for ultrahigh throughput on-bead screening of functional proteins.Nucleic acids research · 2020Article
- Engineering Robust Cellulases for Tailored Lignocellulosic Degradation Cocktails.International journal of molecular sciences · 2020Review
- Evolving a Peptide: Library Platforms and Diversification Strategies.International journal of molecular sciences · 2019Review
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Ultrahigh throughput screening (uHTS) plays an essential role in directed evolution for tailoring biocatalysts for industrial applications. Flow cytometry-based uHTS provides an efficient coverage of the generated protein sequence space by analysis of up to 10(7) events per hour. Cell-free enzyme production overcomes the challenge of diversity loss during the transformation of mutant libraries into expression hosts, enables directed evolution of toxic enzymes, and holds the promise to efficiently design enzymes of human or animal origin. The developed uHTS cell-free compartmentalization platform (InVitroFlow) is the first report in which a flow cytometry-based screened system has been combined with compartmentalized cell-free expression for directed cellulase enzyme evolution. InVitroFlow was validated by screening of a random cellulase mutant library employing a novel screening system (based on the substrate fluorescein-di-β-D-cellobioside), and yielded significantly improved cellulase variants (e.g. CelA2-H288F-M1 (N273D/H288F/N468S) with 13.3-fold increased specific activity (220.60 U/mg) compared to CelA2 wildtype: 16.57 U/mg).
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.