WO2021253362A1 - 一种以烟酰胺为原料制备烟酰胺单核苷酸的方法 - Google Patents
一种以烟酰胺为原料制备烟酰胺单核苷酸的方法 Download PDFInfo
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- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 title claims abstract description 96
- 238000000034 method Methods 0.000 title claims abstract description 67
- 235000005152 nicotinamide Nutrition 0.000 title claims abstract description 48
- 239000011570 nicotinamide Substances 0.000 title claims abstract description 48
- 229960003966 nicotinamide Drugs 0.000 title claims abstract description 48
- 239000002994 raw material Substances 0.000 title claims abstract description 39
- DAYLJWODMCOQEW-TURQNECASA-O NMN(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(O)=O)O2)O)=C1 DAYLJWODMCOQEW-TURQNECASA-O 0.000 title claims abstract 14
- 238000006243 chemical reaction Methods 0.000 claims abstract description 85
- JLEBZPBDRKPWTD-TURQNECASA-O N-ribosylnicotinamide Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](CO)O2)O)=C1 JLEBZPBDRKPWTD-TURQNECASA-O 0.000 claims abstract description 34
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000007788 liquid Substances 0.000 claims abstract description 20
- 238000001471 micro-filtration Methods 0.000 claims abstract description 19
- 239000012528 membrane Substances 0.000 claims abstract description 18
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000001728 nano-filtration Methods 0.000 claims abstract description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims abstract description 12
- IHNHAHWGVLXCCI-FDYHWXHSSA-N [(2r,3r,4r,5s)-3,4,5-triacetyloxyoxolan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H]1OC(C)=O IHNHAHWGVLXCCI-FDYHWXHSSA-N 0.000 claims abstract description 12
- FTVLMFQEYACZNP-UHFFFAOYSA-N trimethylsilyl trifluoromethanesulfonate Chemical compound C[Si](C)(C)OS(=O)(=O)C(F)(F)F FTVLMFQEYACZNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000000872 buffer Substances 0.000 claims abstract description 9
- 239000002904 solvent Substances 0.000 claims abstract description 9
- 101001076781 Fructilactobacillus sanfranciscensis (strain ATCC 27651 / DSM 20451 / JCM 5668 / CCUG 30143 / KCTC 3205 / NCIMB 702811 / NRRL B-3934 / L-12) Ribose-5-phosphate isomerase A Proteins 0.000 claims abstract description 8
- 102000046755 Ribokinases Human genes 0.000 claims abstract description 8
- RAHZWNYVWXNFOC-UHFFFAOYSA-N Sulphur dioxide Chemical compound O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910001425 magnesium ion Inorganic materials 0.000 claims abstract description 7
- 229910021627 Tin(IV) chloride Inorganic materials 0.000 claims abstract description 5
- HPGGPRDJHPYFRM-UHFFFAOYSA-J tin(iv) chloride Chemical compound Cl[Sn](Cl)(Cl)Cl HPGGPRDJHPYFRM-UHFFFAOYSA-J 0.000 claims abstract description 5
- 238000011282 treatment Methods 0.000 claims abstract description 5
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 14
- 239000012295 chemical reaction liquid Substances 0.000 claims description 8
- 238000006555 catalytic reaction Methods 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 239000012510 hollow fiber Substances 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 abstract description 15
- 108090000790 Enzymes Proteins 0.000 abstract description 15
- 239000007787 solid Substances 0.000 abstract description 8
- 230000036632 reaction speed Effects 0.000 abstract description 3
- 230000003197 catalytic effect Effects 0.000 abstract description 2
- 238000007670 refining Methods 0.000 abstract 2
- 239000000243 solution Substances 0.000 description 56
- DAYLJWODMCOQEW-TURQNECASA-N NMN zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)([O-])=O)O2)O)=C1 DAYLJWODMCOQEW-TURQNECASA-N 0.000 description 39
- 239000000047 product Substances 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 238000002360 preparation method Methods 0.000 description 11
- 238000003756 stirring Methods 0.000 description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 6
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 6
- 229950006238 nadide Drugs 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 239000012224 working solution Substances 0.000 description 6
- 239000013067 intermediate product Substances 0.000 description 5
- 230000015556 catabolic process Effects 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005265 energy consumption Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 3
- 235000017557 sodium bicarbonate Nutrition 0.000 description 3
- 239000012535 impurity Substances 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- -1 amide compound Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000007806 chemical reaction intermediate Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000020956 nicotinamide riboside Nutrition 0.000 description 1
- 239000011618 nicotinamide riboside Substances 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000011514 reflex Effects 0.000 description 1
- 239000013557 residual solvent Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/048—Pyridine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
Definitions
- the present invention relates to the technical field of preparation of nicotinamide mononucleotide, in particular to a method for preparing nicotinamide mononucleotide using nicotinamide as a raw material by means of a combination of chemical synthesis and biological enzyme catalysis.
- Nicotinamide mononucleotide is a biochemical substance inherent in biological cells. After being adenylated by nicotinamide nucleotide adenosyl transferase, it is transformed into biological cells that depend on survival.
- the important substance nicotinamide adenine dinucleotide (abbreviated as NAD, also known as coenzyme I, exists in every cell and participates in thousands of reactions).
- NAD nicotinamide adenine dinucleotide
- NMN is the direct precursor of NAD and is currently the most ideal way to supplement NAD. Its level in biological cells directly affects the concentration of NAD, plays an important role in biological cell energy production, and is not harmful to the human body.
- nicotinamide mononucleotide has many pharmacological activities such as regulating immunity, regulating insulin secretion, and affecting the expression of mRNA. It has been developed for medical care such as delaying aging, improving symptoms of Parkinson’s and other senile diseases. , Other uses are also under study. With the increase in people's awareness of the medicinal and health effects of nicotinamide mononucleotide, and the wide application of nicotinamide mononucleotide as a reaction substrate in the chemical industry, the demand for nicotinamide mononucleotide in the market is increasing day by day.
- Nicotinamide also known as nicotinamide, is an amide compound of niacin, which belongs to the precursor substance of NMN.
- nicotinamide as a raw material is a commonly used method for preparing nicotinamide mononucleotide. This method generally adopts the chemical synthesis route shown in the following figure, that is, first convert nicotinamide into the intermediate nicotinamide ribose ( Nicotinamide riboside, referred to as NR), and then convert nicotinamide ribose into the final product nicotinamide mononucleotide.
- NR nicotinamide riboside
- the purpose of the present invention is to solve the technical problem that the refined nicotinamide ribose must be used to complete the conversion into nicotinamide mononucleotide, and at the same time, the more green and environmentally friendly biological enzyme catalytic means In combination with chemical synthesis, a simple and low-cost method for preparing nicotinamide mononucleotide using nicotinamide as a raw material is developed.
- the inventor has carried out a large number of experiments for a long time, and finally developed a method for preparing nicotinamide mononucleotide using nicotinamide as a raw material.
- the method includes the following steps:
- the nicotinamide ribokinase is used to catalyze the reaction of the nicotinamide ribose solution obtained in step 5) at a temperature of 35-39°C, and the pH of the reaction process is controlled to be 7.5 ⁇ 8.0, Nicotinamide mononucleotide is obtained after the reaction is over.
- the reaction in step 1) is carried out at a temperature of 25-35°C.
- the solvent is preferably acetonitrile, because of its good solubility in the reaction, it helps to form a homogeneous system and facilitates the rapid progress of the reaction. , And the by-product ⁇ -isomer is very small ( ⁇ 0.1%).
- the catalyst is preferably trimethylsilyl trifluoromethanesulfonate because of its high reaction stereoselectivity in the reaction. Easy to handle, no metal residues.
- step 1) of the method for preparing nicotinamide mononucleotide using nicotinamide as a raw material provided by the present invention trimethylsilyl trifluoromethanesulfonate or tin tetrachloride, nicotinamide and tetraacetyl ribose
- the molar ratio is 1.2 ⁇ 5:1.2 ⁇ 2:1.
- the second reaction solution is concentrated to remove the solvent.
- the reaction in step 3 is carried out at a temperature of -10 to -5°C .
- step 3 of the method for preparing nicotinamide mononucleotide using nicotinamide as a raw material provided by the present invention the molar ratio of sodium methoxide to tetraacetylribose is 1 to 5:1.
- the temperature of the reaction liquid is maintained at -10 ⁇ -5 in the process of step 4). °C.
- the pore size of the microfiltration membrane used in the microfiltration process is 0.2-1 ⁇ m to remove microorganisms in the reaction solution.
- the nanofiltration process uses hollow fiber membranes with a molecular weight cut-off of 150-250 to remove nicotinamide, residual solvents and most of the inorganic salt impurities in the reaction solution.
- step 6 the amount of each substance added in the entire enzyme-catalyzed reaction system is: Mg ion 10-50mM, ATP 10- 30mM, buffer 20-100mM, nicotinamide ribose 9-27mM, nicotinamide ribokinase 0.2-1g/L.
- the Mg ion is MgCl 2 .
- the buffer is K 2 HPO 4 buffer.
- the preparation method of nicotinamide mononucleotide uses nicotinamide and tetraacetylribose as starting materials, and nicotinamide ribose as the reaction intermediate.
- the whole process avoids the prior art. It is necessary to refine the nicotinamide ribose by crystallization and other methods to participate in the technical requirements of the subsequent conversion into nicotinamide mononucleotide, so the purification steps for nicotinamide ribose are reduced, the process is more streamlined, and the operation is simpler. Lower cost and shorter time-consuming.
- the light yellow liquid obtained above is dissolved in water, and then sent to the membrane concentration equipment for microfiltration and nanofiltration treatments in sequence.
- the microfiltration membrane used in the microfiltration process has a pore size of 0.2 ⁇ m, and the nanofiltration process uses a hollow with a molecular weight cut-off of 150.
- the fibrous membrane is used to obtain a nicotinamide ribose solution (the organic solvent is less than 0.5%), and the obtained solution is ready for use.
- the light yellow liquid obtained above is dissolved in water, and then sent to the membrane concentration equipment for microfiltration and nanofiltration treatments in sequence.
- the microfiltration membrane used in the microfiltration process has a pore size of 0.5 ⁇ m, and the nanofiltration process uses a hollow with a molecular weight cut-off of 200.
- the fibrous membrane is used to obtain a nicotinamide ribose solution (the organic solvent is less than 0.5%), and the obtained solution is ready for use.
- the light yellow liquid obtained above is dissolved in water, and then sent to the membrane concentration equipment for microfiltration and nanofiltration treatments in sequence.
- the microfiltration membrane used in the microfiltration process has a pore size of 0.7 ⁇ m, and the nanofiltration process uses a hollow with a molecular weight cut-off of 200.
- the fibrous membrane is used to obtain a nicotinamide ribose solution (the organic solvent is less than 0.5%), and the obtained solution is ready for use.
- the light yellow liquid obtained above is dissolved in water, and then sent to the membrane concentration equipment for microfiltration and nanofiltration in sequence.
- the microfiltration membrane used in the microfiltration process has a pore size of 1 ⁇ m, and the nanofiltration process uses hollow fibers with a molecular weight cut-off of 250. Film, obtain nicotinamide ribose solution (organic solvent soluble residue is less than 0.5%), and the obtained solution is ready for use.
- the NR solution prepared in the embodiment of the present invention has a faster initial speed in the reaction and requires a lower amount of enzyme.
- the NR solution prepared in the examples of the present invention has a faster initial speed in the reaction, and can avoid the occurrence of the factors shown in the commercially available NR solids-batch 2. A situation where certain impurities are present and the reaction is inhibited.
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- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
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- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Saccharide Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
实验 | pH值 | 温度 | 放置时间 | 稳定性 |
1 | 3 | 2~4℃ | 45天 | 基本不降解 |
2 | 4 | 2~4℃ | 45天 | 降解2.4% |
3 | 5 | 2~4℃ | 45天 | 降解3.7% |
温度 | 甲醇钠用量 | 反应时间 | 反应结果 |
0~5℃ | 2.0eq | 0.5h | 反应完全,产品降解9.2% |
-5~0℃ | 2.0eq | 1.0h | 反应完全,产品降解19.6% |
-10~-5℃ | 2.0eq | 1.5h | 反应完全,产品基本不降解 |
-15~-10℃ | 2.0eq | 5.0h | 反应完全,产品基本不降解 |
甲醇钠用量 | 反应温度 | 反应时间 | 反应结果 |
1.2eq | -10~-5℃ | 24.0h | 反应完全,产品基本不降解 |
1.5eq | -10~-5℃ | 2.0h | 反应完全,产品基本不降解 |
2.0eq | -10~-5℃ | 1.5h | 反应完全,产品基本不降解 |
3.0eq | -10~-5℃ | 1.0h | 反应完全,产品基本不降解 |
5.0eq | -10~-5℃ | 40min | 反应完全,产品基本不降解 |
实验 | 储存温度 | pH值 | 储存时间 | 稳定性 |
1 | 0~4℃ | 3 | 45天 | 基本不降解 |
2 | 0~4℃ | 4 | 45天 | 降解1.3% |
3 | 0~4℃ | 5 | 45天 | 降解2.1% |
Claims (10)
- 一种以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于,所述方法包括如下步骤:1)在乙腈、二氯甲烷、1,2-二氯乙烷或者液态二氧化硫溶剂中,用三氟甲磺酸三甲基硅酯或四氯化锡催化烟酰胺和四乙酰核糖于20~40℃的温度下进行反应,得第一反应液;2)向第一反应液中加入碳酸氢钠、碳酸钠或氢氧化钠,调节pH值为3~5,得第二反应液;3)向第二反应液中加入甲醇钠溶液,于-15~5℃的温度下进行反应,得第三反应液;4)向第三反应液中加入盐酸,调节pH值为3~5,得第四反应液;5)用膜浓缩设备对第四反应液依次进行微滤和纳滤处理,得烟酰胺核糖溶液;6)在Mg离子、ATP和缓冲液存在的环境中,于35~39℃的温度下,用烟酰胺核糖激酶催化步骤5)得到的烟酰胺核糖溶液进行反应,控制反应过程的pH值为7.5~8.0,反应结束后即得烟酰胺单核苷酸。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤1)的反应在25~35℃的温度下进行。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤1)中,所述三氟甲磺酸三甲基硅酯、所述烟酰胺与所述四乙酰核糖的摩尔比为1.2~5:1.2~2:1。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤3)的反应在-10~-5℃的温度下进行。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤3)中,所述甲醇钠与所述四乙酰核糖的摩尔比为1~5:1。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤4)的过程保持反应液的温度为-10~-5℃。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方 法,其特征在于:所述步骤5)中,所述微滤过程采用的微滤膜孔径为0.2~1μm,所述纳滤过程采用截留分子量为150~250的中空纤维膜。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于所述步骤6)中,整个酶催化反应体系中各物质的加入量为:Mg离子10~50mM,ATP 10~30mM,缓冲液20~100mM,烟酰胺核糖9~27mM,烟酰胺核糖激酶0.2~1g/L。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤6)中,所述Mg离子为MgCl 2。
- 根据权利要求1所述的以烟酰胺为原料制备烟酰胺单核苷酸的方法,其特征在于:所述步骤6)中,所述缓冲液为K 2HPO 4缓冲液。
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