ResearchConstruction of Novel Protease Variants
Protein engineering and directed evolution strategies will be exploited to create and screen for novel thermolysin-like protease (TLP) variants that can be used in protein hydrolysis and synthesis applications. TLPs are produced by several members of the bacterial genus Bacillus and several family members are being applied in industrial applications. Broader use of these enzymes is hampered by insufficient stability, specificity or activity under the appropriate conditions.
For the protein engineering approach molecular modeling and available expertise within IMEnz Bioengineering BV will be exploited to improve specific properties. To facilitate directed evolution strategies, regulated and/or inducible expression systems for thermolysin-like proteases in Escherichia coli and Lactococcus lactis will be developed. By means of new high-throughput screening procedures, novel properties, e.g. altered substrate specificity, pH optimum, or improved stability in organic solvents, shall be selected for. Availability of large numbers of TLP variants, with different enzymatic properties, will result in new industrial applications and improve existing processes. A recently constructed TLP variant, that is resistant to boiling, will serve as stable scaffold for these new variants.
Furthermore, IMEnz Bioengineering BV will undertake efforts to improve immobilization efficiencies for TLPs. Immobilized enzymes are advantageous because they manifest superior stability and can easily be recovered from the reaction process, thus enabling longer usage. In particular, peptide/ester synthesis will benefit from immobilized TLP variants, as the substrates are small and size limitation of enzyme-substrate interaction is not likely to occur. It is expected that this project will result in improved biocatalysts for many applications.
If you would like to discuss your specific desires in thermolysin-like proteases, please contact us at the address below.Dr. Bertus van den Burg
Manager Enzyme Technology
IMEnz Bioengineering BV
tel: (+31) 50 3132025