ArticlePloS one2017
Enzyme engineering: A synthetic biology approach for more effective library generation and automated high-throughput screening.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Artificial intelligence and automation in enzyme engineering: evolution, advances, and future perspectives.Bioresources and bioprocessing · 2026Review
- Biosensor-driven strain engineering reveals key cellular processes for maximizing isoprenol production inScience advances · 2025Article
- Synthetic Lipid Biology.Chemical reviews · 2025Review
- Hybrid Exb/Mot stators require substitutions distant from the chimeric pore to power flagellar rotation.Journal of bacteriology · 2024Article
- A growth-based screening strategy for engineering the catalytic activity of an oxygen-sensitive formate dehydrogenase.Applied and environmental microbiology · 2024Article
- Recent advances in microbial and enzymatic engineering for the biodegradation of micro- and nanoplastics.RSC advances · 2024Review
- Tuning Selectivity in CalA Lipase: Beyond Tunnel Engineering.Biochemistry · 2023Article
- New Trends in Bioremediation Technologies Toward Environment-Friendly Society: A Mini-Review.Frontiers in bioengineering and biotechnology · 2021Review
- Modelling of substrate access and substrate binding to cephalosporin acylases.Scientific reports · 2019Article
- Golden Mutagenesis: An efficient multi-site-saturation mutagenesis approach by Golden Gate cloning with automated primer design.Scientific reports · 2019Article
- Holistic engineering of Cal-A lipase chain-length selectivity identifies triglyceride binding hot-spot.PloS one · 2019Article
- Darwin Assembly: fast, efficient, multi-site bespoke mutagenesis.Nucleic acids research · 2018Article
- Highly efficient one-step scarless protein tagging by type IIS restriction endonuclease-mediated precision cloning.Biochemical and biophysical research communications · 2017Article
- The evolving art of creating genetic diversity: From directed evolution to synthetic biology.Biotechnology advancesReview
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Golden Gate strategy entails the use of type IIS restriction enzymes, which cut outside of their recognition sequence. It enables unrestricted design of unique DNA fragments that can be readily and seamlessly recombined. Successfully employed in other synthetic biology applications, we demonstrate its advantageous use to engineer a biocatalyst. Hot-spots for mutations were individuated in three distinct regions of Candida antarctica lipase A (Cal-A), the biocatalyst chosen as a target to demonstrate the versatility of this recombination method. The three corresponding gene segments were subjected to the most appropriate method of mutagenesis (targeted or random). Their straightforward reassembly allowed combining products of different mutagenesis methods in a single round for rapid production of a series of diverse libraries, thus facilitating directed evolution. Screening to improve discrimination of short-chain versus long-chain fatty acid substrates was aided by development of a general, automated method for visual discrimination of the hydrolysis of varied substrates by whole cells.
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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.