Hydrolase Engineering by Circular Permutation

Hydrolase Engineering by Circular Permutation Stefan Lutz, Emory University, Department of Chemistry, Atlanta, GA 30322 Enzyme engineering of lipase B from Candida antarctica (WT CALB) by circular permutation (CP) results in biocatalysts with rate enhancements of up to 100-fold while preserving the high parental enantioselectivity. A recent studies with one variant (cp283∆7) demonstrates the enzymes superior performance as inter and transesterification catalysts for triglycerides and oils in biodiesel production. canola oil / 1-butanol

B.) % product formation

% product formation

A.)

7x

corn oil / 1-butanol

5.5x

1x

1x

C.)

canola oil / ethyl acetate

4x

1x

 cp283∆7  WT CALB

D.)

% product formation

time (h)

% product formation

Crystal structure of domain-swapped cp283∆7 dimer with suicide inhibitor (PDB: 3icw)

time (h)

spent cooking oil / 1-butanol

5x

1x

time (h)

time (h)