Services overview
SERVICES: CUSTOM PROCESS DESIGN
Our strategy: providing solution by combining experimental and computational methods
Braiuca P. et al, TRENDS in Biotechnol., 2006, 24, 419
ENZYME IMMOBILISATION: ANALYSIS OF ENZYME STRUCTURE (HYDROPHILIC/HYDROPHOBIC INTERACTIONS AND ANALYSIS OF AMINO ACIDS INVOLVED IN IMMOBILISATION)
Molecular simulations are used to calculate hydrophobic/hydrophilic balance of enzyme surface. The mapping of hydrophobic and hydrophilic regions provides information on the interaction the protein will establish with the carrier. This information is used to design the immobilization conditions to ensure an appropriate enzyme orientation, maximize the stability of interaction with the carrier and ultimately optimize the performances of the immobilized enzyme.
The location of aminoacid residues utilizable for covalent linkage with the carrier is analysed to choose the most appropriate immobilization chemistry. Particular attention is given to the relative position of the covalent attach points and the active site entrance to maximize substrate accessibility after enzyme-carrier bonds are formed.
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| Lipase CALB | PGA from E. coli | |
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| Trypsin | Thermolysin | |
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| Subtilisin | ||
Basso A. et al., Adv. Synth. Catal, 2007, 349, 877
Degrassi G. et al., Biotechnol. Bioeng., 2006, 93, 344
ENZYME THERMOSTABILITY PREDICTION
A detailed analysis of the enzyme surface by in silico calculation of physical chemical molecular descriptors can be used for the prediction of enzyme (thermo)stability, or to get information about how to improve stability. 3D-QSAR models of mesostable, thermostable and hyperthermostable enzymes are available. It is also possible to study the effect of enzyme glycosylation on thermostability. New enzyme class specific 3D-QSAR models can be designed combining molecular simulations and experimental measurements for the prediction of any property of interest.
Braiuca P. et al., Biotechnol. J., 2007, 2, 214
ENZYME SELECTIVITY PREDICTION
Computational techniques involving molecular modeling coupled with multivariate statistical analysis are used to evaluate and predict quantitatively the substrate selectivity and enantioselectivity of enzymes.
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Example: conformation of tetrahedral intermediates of a) D-PheOMe (c) and L-PheOMe and b) D-AlaOMe (a) and L-AlaOMe
Basso A. et al., J. Mol. Catal. B: Enzymatic, 2002, 19–20, 423
Example: conformation of amide enantiomers in the active site of CALB
Braiuca et al., Adv. Synth. Catal., 2009, 351, 1293
PREDICTING MUTAGENESIS SITES
Prediction of selectivity changes upon mutagenesis of aminoacids in two different enzyme structures
Braiuca P. et al., Biotechnol. Prog., 2004, 20, 1025
Braiuca P. et al., ChemBioChem., 2003, 4, 615
