cover illustration Cutinase from Fusarium solani pisi was subjected to complete site-saturation mutagenesis to scan its entire amino acid sequence and identify those amino acids which contribute to increase its activity and stability towards the anionic detergent AOT. Five different “hot spot” regions were identified as being important in the stabilisation of cutinase towards AOT: (1) The loop near the N terminus (blue); (2) the binding loop 40 52 (bordeaux); (3) the binding loop 171-191 (orange); (4) the loop 151-166 (green); and (5) the large hydrophobic crevice that presumably renders cutinase susceptible to unfolding by anionics (red).This crevice is created when helices encompassing residues 191-211 and 51-63 move apart in the course of unfolding. A kink motion between residues 51-55 and a conformational change in the loop encompassing residues 64-66 characterises this motion. Of the variants that retained catalytic activity similar to native cutinase, variant S54D showed the best stability towards AOT. For further details please see Brissos et al., pp. 387-393.
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