Development of protein factory infrastructure for the production of immobilized biocatalysts
The production of proteins with catalytic activity is a key element of sustainable development. Immobilization engineering allows development of novel immobilization methods that increase the usability of enzymes. The aim of this project is to improve the production and immobilization of catalytic proteins (lipases, transaminases, and dUTPases) at pilot lab level.
The main objective of the infrastructure development project is to recombinantly produce and subsequently immobilize proteins with important biocatalytic activity on lab / pilot-lab scale. Aims of the project are the production and subsequent purification of three families of enzymes (lipases, transaminases, and dUTPases) on a scale of 1-20 L after recombinant production and purification. The bottleneck of these processes is the post-fermentation cell disruption, which requires the purchase of a cell homogenizer which can be operated in a batch or continuous mode within the project in addition to the operation costs.
After recombinant production of the three enzyme families (lipases, e.g. from Candida antarctica or Geobacillus stearothermophilus; transaminases, such as from Arthrobacter or Sinorhizobium sp .; and dUTPases, e.g., from human or E. coli) (gene synthesis, cloning, expression optimization, recombinant expression) and their scale-up to 1-20 L, in the development of novel enzyme immobilization methods we would like to utilize the knowledge and infrastructure of the Institute for Materials and Environmental Chemistry of the Hungarian Academy of Sciences in the development and characterization of heterogeneous catalysts. We intend to co-operate with Dr. Beáta G. Vértessy in applications of dUTPases.
In the course of the work, we plan the combinatorial surface functionalization of carbon and Si, Ti, Al-based nanostructures with mixtures of functionalizing reagents containing amino groups using 16-channel MW autoclave and experimental design methods. The resulting nanostructured supports are used to immobilize the above-mentioned enzymes.
László Poppe, András Tompos, Áron Németh