Enzymatic systems and methods for synthesizing nicotinamide mononucleotide and nicotinic acid mononucleotide
AssigneesNewSouth Innovations Pty Limited, Metro Biotech, LLC
InventorsLindsay Wu, David A. Sinclair, Kyle Meetze
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Abstract
Enzyme-based systems and methods for synthesizing the NAD precursors NMN and NaMN are disclosed. Such methods and systems utilize a mutated form of phosphoribosylpyrophosphate synthetase (PRS) that is superactive and/or other enzyme or enzyme combinations that are immobilized onto a solid surface. The methods and systems substantially increase the efficiency and yield of NAD precursor synthesis.
Claims75
A system for synthesizing a nicotinamide adenine dinucleotide (NAD) precursor, the system comprising a superactive phosphoribosylpyrophosphate synthetase (PRS) mutant, wherein the PRS mutant is less sensitive to the product of the reaction that it catalyzes than a wild type PRS.
The system of any of claims 10-13, wherein the NAMPT or NAPRT is recombinantly produced, isolated, or purified.
A method for synthesizing an nicotinamide adenine dinucleotide (NAD) precursor comprising contacting ribose-5-phosphate with a superactive phosphoribosylpyrophosphate synthetase (PRS) mutant in the presence of adenosine triphosphate (ATP), wherein the PRS mutant is less sensitive to the product of the reaction that it catalyzes than a wild type PRS, and whereby phosphoribosyl pyrophosphate (PRPP) is produced.
A system for synthesizing nicotinamide mononucleotide (NMN), the system comprising nicotinamide riboside kinase (NRK) immobilized onto a surface.
A method for synthesizing nicotinamide mononucleotide (NMN), the method comprising contacting nicotinamide riboside kinase (NRK) immobilized onto a surface with nicotinamide riboside in the presence of adenosine triphosphate (ATP), whereby NMN is produced.
A system for synthesizing nicotinamide mononucleotide (NMN), the system comprising the following enzymes immobilized onto a surface:
- (a) a superactive phosphoribosylpyrophosphate synthetase (PRS) mutant, wherein the PRS mutant is less sensitive to the product of the reaction that it catalyzes than a wild type PRS;
- (b) hexokinase;
- (c) glucose-6phosphate dehydrogenase;
- (d) gluconolactonase;
- (e) 6-phospho gluconate dehydrogenase;
- (f) ribulose-5-phosphate isomerase;
- and
- (g) nicotinamide phosphoribosyl transferase.
Dependencies are inferred from the claim text.