Methods for enzyme library creation: which one will you choose? A guide for novices and experts to introduce genetic diversity

Alejaldre, Lorea; Pelletier, Joelle N.; Quaglia, Daniela

Abstract

Enzyme engineering allows to explore sequence diversity in search for new properties. The scientific literature is populated with methods to create enzyme libraries for engineering purposes, however, choosing a suitable method for the creation of mutant libraries can be daunting, in particular for the novices. Here, we address both novices and experts: how can one enter the arena of enzyme library design and what guidelines can advanced users apply to select strategies best suited to their purpose? Section I is dedicated to the novices and presents an overview of established and standard methods for library creation, as well as available commercial solutions. The expert will discover an up-to-date tool to freshen up their repertoire (Section I) and learn of the newest methods that are likely to become a mainstay (Section II). We focus primarily on in vitro methods, presenting the advantages of each method. Our ultimate aim is to offer a selection of methods/strategies that we believe to be most useful to the enzyme engineer, whether a first-timer or a seasoned user.

• Journal Article Type: Article
• Acceptance Date: Jun 2, 2021
• Online Publication Date: Jul 15, 2021
• Publication Date: 2021-08
• Deposit Date: Jul 1, 2021
• Publicly Available Date: Jul 16, 2022
• Journal: BioEssays
• Print ISSN: 1521-1878
• Electronic ISSN: 1521-1878
• Peer Reviewed: Peer Reviewed
• Volume: 43
• Issue: 8
• Article Number: 2100052
• DOI: https://doi.org/10.1002/bies.202100052
• Public URL: https://nottingham-repository.worktribe.com/output/5750044
• Publisher URL: https://onlinelibrary.wiley.com/doi/10.1002/bies.202100052#