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IMEnz Bioengineering Research 
Biocatalyst Engineering Living organisms are dependent on the enormous diversity of molecular functions that are carried out by proteins. The majority of the molecular reactions occurring within a cell are catalysed by enzymes (biocatalysts). These enzymes are capable of performing reactions with high conversion rates and specificity.

Commercial exploitation of biocatalysts is of interest, since, contrary to the racemic mixtures often obtained with chemical conversions, biocatalytic processes can yield stereo-specific enantiomers and far less by-products. However, natural biocatalysts are often not optimally suited for industrial application. IMEnz Bioengineering has great expertise in improving biocatalyst performance by means of protein engineering and directed evolution.

Protein engineering is the design and construction of mutant proteins, which requires a multidisciplinary approach, involving recombinant DNA techniques, biochemical techniques, protein crystallography, and computer graphics (molecular modeling). IMEnz Bioengineering has applied protein engineering to improve the function and stability of starch-converting enzymes and proteases. For these enzymes, the structural features involved in stability, substrate specificity, pH optimum, catalytic mechanism and product inhibition have been studied and modified.

The directed evolution approach can be the method of choice for the improvement of biocatalysts, in particular for biocatalysts for which structural data are lacking. Random mutagenesis procedures are applied to create libraries of biocatalyst variants. Subsequently, specific screening procedures are developed to select variants with desired properties.
IMEnz Bioengineering has ample experience in biocatalyst engineering and can be your partner for the improvement of biocatalyst function, stability and applications. In addition, services for biocatalyst identification, cloning, expression and production are offered.
If you would like to discuss your specific desires in biocatalist engineering, please contact us.

IMEnz Bioengineering Research
Construction 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.

IMEnz Bioengineering Research
Enzyme Libraries for Application in
Industrial and Pharmaceutical Biotransformation

IMEnz Bioengineering BV provides access to unique libraries of enzyme variants and conducts the screening of these libraries for the selection of biocatalysts with desired properties.

The first library that has been made available contains more than 350 protease variants. The individual biocatalysts exhibit a large variation with respect to stability, pH optimum, substrate specifcity, metal ion dependence, and absolute activity, both in aqueous and in organic environments. This particular class of proteases has been shown to be applicable for many kinds of bioconversion reactions, as diverse as hydrolysis and synthesis. The stereoselectivity of transformations obtained by biocatalysts makes them extremely powerfull for chiral resolution applications.

IMEnz Bioengineering BV utilizes the library for the selection and identification of enzymes with desired or novel properties.

To that goal, IMEnz Bioengineering BV offers the following services:

  • development of application-oriented screening procedures
  • screening of the libraries for specific biotransformations
  • identification of biocatalysts involved
  • isolation, production and upscaling of identified, interesting variants
  • further improvement of selected biocatalysts by means of protein engineering and or directed evolution
  • assistance in obtaining exclusive rights to use selected variants
  • assistance in search and operation of new libraries

Other libraries of industrial enzymes, including chitinases, esterase, lipases and sugar-modifying enzymes are or will become available in due time also.