The present disclosure provides modified looped proteins comprising at least one looped region, wherein the at least one looped region comprises a cell penetrating peptide (CPP). In some embodiments, the present disclosure provides polynucleotides encoding the modified looped proteins and methods for their production.

This disclosure relates to methods of treating cancer or initiating, enhancing, or prolonging an anti-tumor response in a subject in need thereof comprising administering to the subject an effective amount of a checkpoint inhibitor in combination with a purine cleaving enzyme or a vector encoding expression thereof, and a prodrug cleaved by said purine cleaving enzyme. In certain embodiments, this disclosure relates to methods of treating cancer or initiating, enhancing, or prolonging an anti-tumor response in a subject in need thereof comprising administering to the subject an effective amount of a checkpoint inhibitor in combination with a purine cleaving enzyme, or a vector encoding expression thereof, in the absence of a prodrug cleaved by said purine cleaving enzyme.

The present invention provides engineered purine nucleoside phosphorylase (PNP) enzymes, polypeptides having PNP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing PNP enzymes are also provided. The present invention further provides compositions comprising the PNP enzymes and methods of using the engineered PNP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

The present invention provides engineered purine nucleoside phosphorylase (PNP) enzymes, polypeptides having PNP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing PNP enzymes are also provided. The present invention further provides compositions comprising the PNP enzymes and methods of using the engineered PNP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

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 invention provides a method of determining a predisposition to a condition associated with ATP depletion, such as an ischaemic event, in a subject comprising: a. measuring the concentration of one or more purines in a body fluid of the subject, the purines being selected from adenosine, inosine, hypoxanthine, xanthine and ATP, and b. comparing the measured concentration with a threshold concentration of the one or more purines, wherein the threshold concentration is preferably in the range 2 [micro]M to 8 [micro]M and wherein a measured concentration higher than the threshold concentration indicates the presence of ischaemia.

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 invention provides engineered purine nucleoside phosphorylase (PNP) enzymes, polypeptides having PNP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing PNP enzymes are also provided. The present invention further provides compositions comprising the PNP enzymes and methods of using the engineered PNP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

Provided are an siRNA which inhibits purine nucleoside phosphorylase (PNP) gene expression, a pharmaceutical composition comprising the siRNA, and an siRNA conjugate, capable of effectively treating and/or preventing abnormal uric acid metabolism, or diseases or physiological conditions caused by abnormal uric acid metabolism. Each nucleotide in the siRNA is independently a modified or unmodified nucleotide.

The present invention relates to the field of biotechnologies. Disclosed are an enzyme composition for preparing ?-nicotinamide mononucleotide (NMN), and an application thereof. In the present invention, by using adenosine and nicotinamide as raw materials, D-ribose 1-phosphate and a nicotinamide ribose intermediate are generated under enzyme catalysis of an enzyme composition of a PNP enzyme and an NRK enzyme, and finally NMN is obtained; only two enzymes need to be involved in the whole reaction system, and the by-product adenine can be recycled. Moreover, only one molecule of ATP needs to be consumed for generating one molecule of NMN, thereby greatly reducing the process cost. The reaction of synthesizing NMN by NRK enzyme catalysis in the last step is irreversible, and thus, the substrate conversion rate can be greatly improved and the production cost can be further reduced.