TH36623A - - Google Patents

Abstract

DC60 (09/09/54) [Issue] A method for the production of nucleoside-5'-phosphate ester (nulceoside-5'-phosphate ester) was provided for a non-cost basis. It is expensive and efficient by adding phosphoril groups to biochemical nucleosides [Key point] nucleoside-5-phosphate. Esters are produced by allowing composite phosphatase, especially asidphosphatase to have increased affinity for nucleosides. And / or stability against Increased temperatures, and microorganisms, which were induced by genes that encoded the active protein of the anhydrous phosphatase to operate under pH conditions of 3.0 to 5.5 on the nucleos. Sites and phosphate groups which are Choose from a group that contains polyphosphoric acid. Or its salts, phenylphosphoric acid or its salts, and carbamyl phosphate or its salts to produce nucleoside-5'-phosphate ester. And the collection of that [problem] method for the production of nucleoside-5'-phosphate. An ester (nucleoside-5'-phosphate ester) is provided inexpensively and effectively by adding a phosphoril group to a biochemical nucleoside. Important] Nucleoside-5-Phosphate Esters are produced by allowing composite phosphatase, especially asidphosphatase to have increased affinity for nucleosides. And / or stability against Temperature rise, and Microorganisms, which were induced by genes encoding the active protein of the anidphosphatase to operate under pH conditions 3.0 to 5.5 on nucleosides and phosphate groups. Which was Choose from a group that contains polyphosphoric acid. Or its salts, phenylphosphoric acid or its salts, and carbamyl phosphate or its salts to produce nucleoside-5'-phosphate ester. And collecting that

Claims (5)

Disclaimer (in total), which will not appear on the advertisement page: 1. It is mutated only at the 74th glycine part and / or 153 isoleucine; (c) the amino acid sequence. SEQ ID NO: 22,24, or 26, minos, mutated only at the 92th glycine fraction and / or 171 isoleucine; And (d) the amino acid sequence of SEQ ID NO: 28, which was mutated only at the 88th glycine portion and / or the 167th isoleucine. 2. Method for nucleus production. Oxide-5 \ '- Phosphate Esters that comprise the phases of It allows microorganisms to operate under pH 3.0 to 5.5 on nucleosides and phosphate groups to produce nucleoside-5 \ '- phosphate ester and their collection. Where the microorganism is reproduced with recombinant DNA composed of the genes encoding azidphosphatase, as defined in claim 1 3. Method of claim 1. Or 2 where Km for the nucleoside below 100 4. Method according to one of the preceding claims, wherein asydophosphatase remains active Viti that After being left at 50 ํ for 30 minutes at pH 7.0 5. Method according to one of the previous claims, wherein asymmetric asidophosphatase had a higher degree of resistance than the original variety. image The nucleosides were obtained from bacteria of the genus Escherichia, the genus Morganella, the genus Providencia, the Enterobacter, the genus Klebsiella, or the genus Serratia 1. 1. and which were mutated by replacing the glycine part. The 74th and / or the 153 isoleucine and also consists of a The strains were selected from the replacement group for the 63th leucine, the 65th alanine, the 66th glutamic acid, the 69th asparagene, the serie. Na 71, serine 72, serine 85, alanine 92, alanine 94, glutamic acid 104, part 116th aspartic acid, 130th serine, 135th theronine, 136th glutamic acid and / or 151th theronine 8. Method Of claim 6 or claim 7 where the leucin part 63 was replaced with glutamine, the alanine 65 was replaced with glutamine, acid A The 66th glutamic was replaced with alanine, 69th asparagene was replaced with aspartic acid, 71th serine was replaced with alanine. The 72nd serine was replaced with alanine, the 74th glycine part with aspartic acid, the 85th serine was replaced with tyrosine, ala. Nine 92 was replaced with serine, alanine 94 was replaced with glutamic acid, 104th glutamic acid was replaced. With the glycine part, the aspartic acid 116 was replaced with the glutamic acid, the 130 th serine was replaced with the glutamic acid, the theronine at 135 was replaced with lysine, 136th glutamic acid was replaced with glutamic acid, 135th theronine was replaced with lysine, alanine and /. Or the 153th isoleucine was replaced with theronine 1.a and with the mutation, replacing the 118th glutamic acid portion and / or the mutation which replaces the 165th theronine 1 0. Method of claim 9 where the glutamic acid portion is replaced by the glycine and / or the aforementioned theronine. Replaced by alanine part 1 1. Method of any of the preceding claims. Where the aforementioned phosphate group was selected From the group containing polyphosphoric acid or its salts, phenylphosphoric acid or its salts, acetylphosphoric acid Or the salts of that substance and carbamyl phosphates or salts of that substance 1 2. Asydophosphatase mutant, as defined in any of the Clause 1 or 3 or 10 1. 3. The decoded stand for Asidphosphates as defined in Clause 12 1. 4. Recombinant DNA containing genes as defined in claim 13 1. 5. Existing microorganisms with recombinant DNA as defined in claim 14.