Protein engineering

Protein engineering is the process of developing useful proteins through mutagenesis approaches.

 

Rational enzyme engineering and reverse engineering.

 

Figure 1. Conserved salt bridges in lipolytic enzyme family XIII.2. Phylogenetic tree of lipolytic enzymes from family XIII and XIII.2 showing the conservation of an interloop salt bridge located in i -2 and i – 4 from the catalytic Asp and His respectively (left). Tridimensional structure of EstGtA2 and location of the family XIII.2-conserved salt bridges studied by combinatorial alanine scanning mutagenesis and location of the conserved interloop salt bridge (right).

Charbonneau DM and Beauregard M. Role of key salt bridges in thermostability of G. thermodenitrificans EstGtA2: Distinctive patterns within the new bacterial lipolytic enzyme family XIII.2PLOS ONE. 2013, 8(10), e76675

 

Directed evolution of PML.

 

Figure 2. Directed evolution of PML. Library screening scheme (left) and structural changes involved in lipase stabilization solved by X-ray crystal structure of Dieselzyme4 (right).

Korman TP, Sahachartsiri B, Charbonneau DM, Huang GL, Beauregard M and Bowie JU. Dieselzymes: development of a stable and methanol tolerant lipase for biodiesel production by directed evolutionBiotechnol Biofuels. 2013, 6:70