Abstract:
Provided herein are systems and components thereof for improving protease activity. The systems make use of an emulsion for in vitro compartmentalization of a library of synthetic compounds, each compound having a gene linked to a protease substrate and selectable marker. Expressed enzymes with greater protease activity will preferentially hydrolyze the protease substrate, whereas enzymes with less protease activity will leave the substrate intact. Removal of the non-hydrolyzed compounds provides an enriched gene library encoding for more active protease variants. Also described are synthetic compounds and emulsions which can be used in the methods.
Abstract:
The present invention relates to polynucleotide constructs for in vitro and in vivo transcription/translation of genes of interest or variants of a gene of interest as well as microorganism host cells comprising such constructs and methods for producing a polypeptide of interest in such microorganism host cells.
Abstract:
Provided herein are methods and means for enhancing lipase activity. The system makes use of an emulsion for in vitro compartmentalization of a library of synthetic compounds which have a polynucleotide linked to a lipase substrate (e.g., a triglyceride). Expressed polypeptides having greater lipase activity will preferentially hydrolyze the substrate from the linked polynucleotide. Genes encoding polypeptides having less lipase activity will remain linked to the substrate and may be removed to enrich the library for more active variants. Also described are synthetic compounds and emulsions which can be used in the methods.
Abstract:
Provided herein are methods and means for enhancing enzymatic activity. The system makes use of an emulsion for in vitro compartmentalization of a library of synthetic compounds which have a gene and a marked enzymatic substrate both directly linked to a solid phase. Expressed enzymes with greater activity will preferentially release the selectable marker from the solid phase, whereas enzymes with less activity will leave the markers intact. Removal of the marked compounds provides an enriched gene library encoding for more active variants. Also described are synthetic compounds and emulsions which can be used in the methods.
Abstract:
The present invention relates to endoglucanase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, expression vectors, and recombinant host cells comprising the polynucleotides; and methods of using the variants.