In vitro evolution of enzymes

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 978; p. 73
• Main Authors: Golynskiy, Misha V, Haugner, 3rd, John C, Morelli, Aleardo, Morrone, Dana, Seelig, Burckhard
• Format: Journal Article
• Language: English
• Published: United States 2013
• Subjects: Directed Molecular Evolution; Enzymes - chemistry; Enzymes - genetics; Enzymes - metabolism; Gene Library; Protein Engineering - methods
• ISSN: 1940-6029
• DOI: 10.1007/978-1-62703-293-3_6

In the past decade, in vitro evolution techniques have been used to improve the performance or alter the activity of a number of different enzymes and have generated enzymes de novo. In this review, we provide an overview of the available in vitro methods, their application, and some general considerations for enzyme engineering in vitro. We discuss the advantages of in vitro over in vivo approaches and focus on ribosome display, mRNA display, DNA display technologies, and in vitro compartmentalization (IVC) methods. This review aims to help researchers determine which approach is best suited for their own experimental needs and to highlight that in vitro methods offer a promising route for enzyme engineering.