An adenosine-involved fully enzymatic synthesis method for NMN includes the steps under the same reaction system: (A) reacting, under the catalytic action of a yeast cell, adenosine, phosphate and sugar that is metabolizable by a yeast cell, so as to generate ATP; and (B) generating NMN by an enzymatic reaction including the step of under the action of NAMPT, reacting nicotinamide, PRPP and ATP to generate NMN, ADP and phosphate. In this way, a series of reactions such as the generation (regeneration) of ATP, the synthesis of NMN and the utilization of ATP are performed in one reaction system in a unified manner, and thus efficient synthesis of NMN can be implemented.

The present disclosure provides an innovative method for improving the enzyme activity of an NMN biosynthetic enzyme Nampt, and relates to the technical field of genetic engineering. A mutant protein of the present disclosure is obtained by firstly analyzing a target protein Nampt using two softwares FoldX and DeepDDG, and then predicting multiple key sites influencing the enzyme functions and finally performing the semi-rational design of the enzyme. In the examples of the present disclosure, 10 mutant strains are constructed using the designed primers according to the principle of point mutation, and 8 of the mutants have higher activity than a wild-type strain, in which the NMN yield of the mutant Nampt-V365L is increased by 62%, and the NMN yields of the mutants Nampt-S248A, Nampt-N164L, Nampt-S382M, Nampt-A245T and Nampt-A208G are increased by 34%, 27%, 27%, 22% and 17% respectively.

Methods and compositions for preventing or treating peripheral neuropathy, cognitive deficits, inactivity, depression, chemotherapy and/or radiotherapy induced peripheral neuropathy and cognitive deficits, and improving cognitive performance, in a subject in need thereof are disclosed. The disclosed methods include the step of administering to the subject an effective amount of an agent that increases the level of NAD.sup.+ in the subject.

Provided herein are pharmaceutical compositions for local administration of metabolic inhibitors, methods of locally administering such compositions, and rapid diagnostic methods for identifying mutant allele during the course of a surgical procedure.

A method of making nicotinamide mononucleotide (NMN), nicotinamide mononucleotide derivatives, or mixtures thereof is disclosed. The method involves the in vitro artificial enzymatic pathways comprised: the generation of alpha-D-ribose-1-phosphate from numerous substrates followed by the synthesis of nicotinamide mononucleotide catalyzed by nicotinamide riboside phosphorylase and nicotinamide riboside kinase or the generation of 5-phospho-alpha-D-ribose-1-diphosphate from nucleotides followed by the synthesis of nicotinamide mononucleotide catalyzed by nicotinamide phosphoribosyltransferase. The multiple enzymes were reconstituted in one pot, wherein in-situ removal of byproducts that can be converted to other non-inhibitory chemicals with supplementary enzymes push the overall biotransformation toward the synthesis of nicotinamide mononucleotide. Furthermore, nicotinamide mononucleotide can be converted to its derivatives—nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate.

The present invention provides a method for preparing nicotinamide mononucleotide (NMN) by bioanalysis. The method includes a step of catalytically reacting a plurality of raw materials including nicotinamide, ATP, and ribose in the presence of nicotinamide phosphoribosyltransferase (Nampt), ribose phosphate pyrophosphokinase, and ribokinase, to prepare the NMN.

Methods and compositions for preventing or treating peripheral neuropathy, cognitive deficits, inactivity, depression, chemotherapy and/or radiotherapy induced peripheral neuropathy and cognitive deficits, and improving cognitive performance, in a subject in need thereof are disclosed. The disclosed methods include the step of administering to the subject an effective amount of an agent that increases the level of NAD.sup.+ in the subject.

An object of the present invention is to provide a method for producing nicotinamide mononucleotide (NMN) with excellent production efficiency. The method for producing NMN according to the present invention (the first aspect) comprises the step of bringing a transformant with enhanced expression of enzymes nicotinamide phosphoribosyltransferase (Nampt), phosphoribosyl pyrophosphate synthetase (Prs) and polyphosphate kinase (Ppk), a cell-free protein synthesis reaction solution having the three enzymes expressed, or a treated product thereof into contact with a mixture containing ribose-5-phosphate (R5P), nicotinamide (NAM), ATP and polyphosphate.

The present disclosure is directed to methods (e.g., in vitro methods) for use of nicotinamide phosphoribosyltransferase (NAMPT) as a biomarker in radiation-induced lung injury (RILI). Provided herein is an in vitro method for the diagnosis, prognosis, and/or monitoring of RILI in a human subject by providing a tissue or plasma sample from the subject and detecting the level of NAMPT therein, wherein a higher level of NAMPT in the tissue or plasma sample from the subject compared to a healthy control or a reference value is indicative for the presence of RILI in the subject. Further provided herein is a method of detecting NAMPT in a human subject by obtaining a biological sample from the subject, detecting the presence of NAMPT in the sample by contacting the sample with a capture agent that specifically binds NAMPT, and detecting binding between NAMPT and the capture agent.

Methods and compositions for preventing or treating peripheral neuropathy, cognitive deficits, inactivity, depression, chemotherapy and/or radiotherapy induced peripheral neuropathy and cognitive deficits, and improving cognitive performance, in a subject in need thereof are disclosed. The disclosed methods include the step of administering to the subject an effective amount of an agent that increases the level of NAD.sup.+ in the subject.