The present disclosure relates to a genetically engineered strain with high production of uridine and its construction method and application. The strain was constructed as follows: heterologously expressing pyrimidine nucleoside operon sequence pyrBCAKDFE (SEQ ID NO:1) on the genome of E coli prompted by strong promoter P.sub.trc to reconstruct the pathway of uridine synthesis; overexpressing the autologous prsA gene coding PRPP synthase by integration of another copy of prsA gene promoted by strong promoter P.sub.trc on the genome; deficiency of uridine kinase, uridine phosphorylase, ribonucleoside hydrolase, homoserine dehydrogenase I and ornithine carbamoyltransferase. When the bacteria was used for producing uridine, 40-67 g/L uridine could be obtained in a 5 L fermentor after fermentation for 40-70 h using the technical scheme provided by the disclosure with the maximum productivity of 0.15-0.25 g uridine/g glucose and 1.5 g/L/h respectively which is the highest level of fermentative producing uridine reported at present.

The present disclosure relates to a novel method, expression vectors, and host cells for producing nicotinamide riboside by regulating the pathways that lead to the production of nicotinamide riboside.

The present disclosure relates to a novel method, expression vectors, and host cells for producing nicotinamide riboside by regulating the pathways that lead to the production of nicotinamide riboside.

The present invention is directed to microbial production of nicotinamide riboside and/or nicotinamide mononucleotide using a genetically modified fungus.

An acid phosphatase mutant, and method for preparing nicotinamide riboside by same. The mutant is a protein of the following (a), (b) or (c): (a) a protein, having an amino acid sequence shown in SEQ ID NO: 3; (b) a protein, derived from (a), having catalytic activity higher than an acid phosphatase parent having an amino acid sequence shown in SEQ ID NO: 2, obtained by substituting, deleting or adding several amino acids in the amino acid sequence shown in SEQ ID NO: 3, using nicotinamide mononucleotide as a substrate; (c) a protein, having catalytic activity higher than the acid phosphatase shown in SEQ ID NO: 2, having 90% or more homology with the amino acid sequence of the protein defined by (a) or (b), and using nicotinamide mononucleotide as a substrate. It is used to prepare nicotinamide riboside and the conversion rate can be 99%.

An acid phosphatase mutant, and method for preparing nicotinamide riboside by same. The mutant is a protein of the following (a), (b) or (c): (a) a protein, having an amino acid sequence shown in SEQ ID NO: 3; (b) a protein, derived from (a), having catalytic activity higher than an acid phosphatase parent having an amino acid sequence shown in SEQ ID NO: 2, obtained by substituting, deleting or adding several amino acids in the amino acid sequence shown in SEQ ID NO: 3, using nicotinamide mononucleotide as a substrate; (c) a protein, having catalytic activity higher than the acid phosphatase shown in SEQ ID NO: 2, having 90% or more homology with the amino acid sequence of the protein defined by (a) or (b), and using nicotinamide mononucleotide as a substrate. It is used to prepare nicotinamide riboside and the conversion rate can be 99%.

The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using an endonuclease V activity and initiator with a 3-penultimate deoxyinosine.

The present invention provides a method for producing a target substance, the biosynthetic pathway of which is ATP-dependent, for example, amino acids, nucleosides, nucleotides, isoprenoids, and peptides, by fermentation of a bacterium which has been modified to overexpress a gene encoding a protein having H.sup.+-translocating membrane-bound pyrophosphatase activity, for example, the hppA gene native to R. rubrum or a variant thereof.

The present invention relates to compositions, kits and methods for making and using RNA compositions comprising in vitro-synthesized ssRNA inducing a biological or biochemical effect in a mammalian cell or organism into which the RNA composition is repeatedly or continuously introduced. In certain embodiments, the invention provides compositions and methods for changing the state of differentiation or phenotype of a human or other vertebrate cell. For example, the present invention provides mRNA and methods for reprogramming cells that exhibit a first differentiated state or phenotype to cells that exhibit a second differentiated state or phenotype, such as to reprogram human somatic cells to pluripotent stem cells.

The present invention relates to compositions, kits and methods for making and using RNA compositions comprising in vitro-synthesized ssRNA inducing a biological or biochemical effect in a mammalian cell or organism into which the RNA composition is repeatedly or continuously introduced. In certain embodiments, the invention provides compositions and methods for changing the state of differentiation or phenotype of a human or other vertebrate cell. For example, the present invention provides mRNA and methods for reprogramming cells that exhibit a first differentiated state or phenotype to cells that exhibit a second differentiated state or phenotype, such as to reprogram human somatic cells to pluripotent stem cells.