Enhancing Coupled Catalytic Activity of Multi-Enzyme Cascades With Liquid-Liquid Phase Separation Using Peptide-Based Condensates
AssigneeThe Government of the United States of America, as represented by the Secretary of the Navy
InventorsJoyce C. Breger, Kimihiro Susumu, Igor L. Medintz, Gregory A. Ellis, Sebastian A. Diaz, Shelby L. Hooe
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Abstract
Enzyme cascade reactions, wherein the product of a first enzyme is the substrate of a second enzyme and so forth, were found to be enhanced by the presence of peptides. It is believed that the peptides operate by forming coacervates, which are membrane-less compartments where, in the case of coacervates formed by peptides, the liquid-liquid phase separation involves water as the continuous phase both inside and outside the coacervate.
Claims9
A method of conducting an enzyme cascade reaction, the method comprising:
- providing a plurality of enzymes configured as an enzymatic cascade wherein the product of a first enzyme is the substrate of a second enzyme and so forth, wherein the enzymatic cascade comprises at least two different enzymes;
- contacting the enzymatic cascade with a substrate of the first enzyme while contacting the enzymatic cascade with a peptide under conditions effective to form a peptide-driven coacervate;
- and
- allowing a reaction to proceed so that each of the plurality of enzymes acts in succession to produce an end product,
- wherein at least one of the enzymes comprises multiple polyhistidine tags acting to cross-link the nanoparticles into the cluster.
A method of conducting an enzyme cascade reaction, the method comprising:
- providing a plurality of enzymes configured as an enzymatic cascade wherein the product of a first enzyme is the substrate of a second enzyme and so forth, wherein the enzymatic cascade comprises at least two different enzymes;
- contacting the enzymatic cascade with a substrate of the first enzyme while contacting the enzymatic cascade with a peptide under conditions effective to form a peptide-driven coacervate;
- and
- allowing a reaction to proceed so that each of the plurality of enzymes acts in succession to produce an end product,
- wherein at least one of the enzymes comprises multiple polyhistidine tags acting to cross-link the nanoparticles into the cluster,
- wherein the plurality of enzymes comprises Bacillus subtilis aspartate ammonia lyase (AspB), E. coli aspartokinase III (LysC), and E. coli aspartate-β-semialdehyde dehydrogenase (Asd).
Dependencies are inferred from the claim text.