Standard protein engineering usually involves the substitution of one or more amino acids by other amino acids chosen from the remaining nineteen common alternatives. While this approach has a proven track record, it is nevertheless extremely limited by the chemical space offered by the 20 canonical amino acids.
Over the last 25 years, techniques have been developed to site specifically introduce non-canonical amino acids (ncAA) into proteins, while using the existing genetic code [1,2]. Advantageously, these approaches provide a means to explore chemical space, create new catalytic opportunities and perform bioorthogonal conjugate reactions or assemblies.
Despite the increasing use of ncAA, this approach has rarely been used to engineer carbohydrate-active enzymes (CAZymes). In work aimed at extending the chemical space of CAZymes, we are studying the production of nCAA-bearing glycoside hydrolases and carbohydrate-binding modules . In this presentation, we will describe current progress towards this goal, focussing both on the methodological aspects and on prospects for the future creation of artificial enzymes displaying new functions or architectures.
This research was supported by the pre-competitive program of Toulouse White Biotechnology (TWB), the Institut National de la Recherche Agronomique (INRA) and the 3BCAR Carnot Institute, through the project INSEREE (2015-2018), the ANS CHIMZYM (2016 & 2017) and the project i-INSEREE (2018-2019) respectively. The iGEM Toulouse INSA-UPS team 2018 (http://2018.igem.org/Team:Toulouse-INSA-UPS) is composed of G. Bordes, A. Pelus, Y. Bouchiba, C. Burnard, J. Delhomme, J. Pérochon, M. Toanen, A. Verdier, C. Wagner, S. Barbe, B. Enjalbert, J. Esque, M.-P. Escudié, R. Fauré, M. Guionnet, A. Henras, S. Heux, P. Millard, C. Montanier, and Y. Romeo.
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- Ramos N.; Bigot D.-J.; Ramillon-Devolve V.; Hebra T.; Montanier C.; Truan G.; Fauré R.; Nouaille S. In preparation