The invention relates to enzyme clusters of resveratrol biosynthesis enzymes that improve the production of resveratrol in bacteria Escherichia coli. Enzyme clusters are formed when enzymes are linked with selected orthogonal pairs of CC peptides, forming dimeric coiled coils. The invention also relates to bacteria E. coli containing enzyme clusters. In the case of resveratrol, it has been shown that the fusion of enzymes with the CC-peptide pairs forming the dimeric coiled coil increases the production of resveratrol. The advantage of using these CC-peptide pairs is that they are orthogonal, i.e. they interact specifically, each CC-peptide only with its designed pair. Thus, we can ensure designed and controlled formation of enzyme clusters for the reactions involving more than two enzymes.
F.33 Slovenian patent
COBISS.SI-ID: 6341402We presented our results at the Slovenian Biochemical Society meeting. Synthetic scaffolds proved to be successful tool for improving biosynthetic pathway efficiency. Our aim was to test the activity of large multi-enzymatic complexes where clustering was mediated by polypeptide elements forming ordered structures. Complexes were produced in Escherichia coli and tested in small scale fermentation bioprocess. Multi-enzymatic particles were purified in very simple isolation process that includes only cellular lysis, centrifugation and final washing and tested in vitro in the production process. Reaction efficiencies of multi-enzymatic particles were compared to reaction systems where soluble enzymes were not linked together or to reaction systems composed of otherwise linked enzymes but not integrated into large subcellular structures.
F.02 Acquisition of new scientific knowledge
COBISS.SI-ID: 5778970At the meeting the results from our research in the field of designed protein origami, e.g. designed self-assembled polypeptide nanostructures from a single chain, and the possibility for their functionalization to improve biocatalysis were presented.
B.04 Guest lecture
COBISS.SI-ID: 5833242