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Biomolecules 2013, 3(4), 778-811; doi:10.3390/biom3040778

Improvement of Biocatalysts for Industrial and Environmental Purposes by Saturation Mutagenesis

Department of Life Sciences and Systems Biology, University of Torino, via Accademia Albertina 13, Torino 10123, Italy
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Received: 21 August 2013 / Revised: 22 September 2013 / Accepted: 23 September 2013 / Published: 8 October 2013
(This article belongs to the Special Issue Enzymes and Their Biotechnological Applications)
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Abstract

Laboratory evolution techniques are becoming increasingly widespread among protein engineers for the development of novel and designed biocatalysts. The palette of different approaches ranges from complete randomized strategies to rational and structure-guided mutagenesis, with a wide variety of costs, impacts, drawbacks and relevance to biotechnology. A technique that convincingly compromises the extremes of fully randomized vs. rational mutagenesis, with a high benefit/cost ratio, is saturation mutagenesis. Here we will present and discuss this approach in its many facets, also tackling the issue of randomization, statistical evaluation of library completeness and throughput efficiency of screening methods. Successful recent applications covering different classes of enzymes will be presented referring to the literature and to research lines pursued in our group. The focus is put on saturation mutagenesis as a tool for designing novel biocatalysts specifically relevant to production of fine chemicals for improving bulk enzymes for industry and engineering technical enzymes involved in treatment of waste, detoxification and production of clean energy from renewable sources.
Keywords: biocatalysis; directed evolution; synthetic biology; protein engineering; industrial biotechnology; bioremediation; fine chemistry; saturation mutagenesis; screening methods biocatalysis; directed evolution; synthetic biology; protein engineering; industrial biotechnology; bioremediation; fine chemistry; saturation mutagenesis; screening methods
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MDPI and ACS Style

Valetti, F.; Gilardi, G. Improvement of Biocatalysts for Industrial and Environmental Purposes by Saturation Mutagenesis. Biomolecules 2013, 3, 778-811.

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