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H. Mansouri Khosravi, C. P. Goncalves, F. Rudroff, S. Feroz, M.D. Mihovilovic:
"Activity And Stability Improvement Of Cyclohexanone Monooxygenase By Protein Engineering";
Poster: ProtStab 2018, vilnius; 16.05.2018 - 18.05.2018.

Enzyme stabiliy is a significant factor in biocatalytic application of enzymes and there is a high demand to provide stable enzyme by different approaches.[1] Cyclohexanone monooxygenase (CHMOAcineto from Acinetobacter sp. NCIMB 9871) is the most well studied Bayer-Villiger monooxygenae and a valuable biocatalyst for industrial application due to it´s impressive substrate scope as well as exquisite chemo, regio and enantioselectivity. But due to low stability of this enzyme the industrial application of this important biocatalyst is hampered. The main objective of this project is to overcome this obstacle by using protein engineeing approach and improve its stability. Iterative rounds of semi-rational protein design was applied to improve CHMOAcineto thermostability. For that, we have used a data-driven protein design method that requires fewer homologous sequences than the traditional consensus approach and utilizes structural information to limit the number of variants created.[3] The combination of single point mutations gave rise to a library of 14 mutants. Both activity and thermostability for all mutants have been evaluated. We have found several single point mutants with increased activity and half-life. After combination, we ended up with 8 mutations containing variant which showed higher activity and 4 mutants were found experimentally to be more thermostable than the wild-type.