CN108998484A - A kind of preparation method of NADP coenzyme - Google Patents
A kind of preparation method of NADP coenzyme Download PDFInfo
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- CN108998484A CN108998484A CN201811016614.3A CN201811016614A CN108998484A CN 108998484 A CN108998484 A CN 108998484A CN 201811016614 A CN201811016614 A CN 201811016614A CN 108998484 A CN108998484 A CN 108998484A
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- starting material
- adenine dinucleotide
- niacinamide
- ribokinase
- nicotinamide
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- 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
- C12P19/36—Dinucleotides, e.g. nicotineamide-adenine dinucleotide phosphate
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Abstract
Disclosed by the invention is a kind of preparation method of NADP coenzyme, when solving in the prior art using biological enzyme preparation nicotinamide adenine dinucleotide, the not high problem of complicated for operation and yield.The present invention includes step 1: obtaining nicotinamide adenine dinucleotide；NADP is made after ATP phosphorylation in nicotinamide adenine dinucleotide by second step；The preparation process of nicotinamide adenine dinucleotide are as follows: starting material, niacinamide ribokinase, nicotinamide mononucleotide adenylase and atriphos are added to reaction in reaction vessel simultaneously and generate nicotinamide adenine dinucleotide；The concentration of the starting material is 30~40g/L, and the ratio that niacinamide ribokinase and nicotinamide mononucleotide adenylase are added is 2~3 ︰ 1~2；The additional amount of the niacinamide ribokinase is the 3~5% of starting material weight；The molar ratio of the atriphos and starting material is 2.1~2.3 ︰ 1.The present invention has many advantages, such as to simplify operation, improves yield.
Description
Technical field
The present invention relates to biological enzyme preparation fields, and in particular to be a kind of NADP coenzyme preparation method.
Background technique
NADP is nicotinamide-adenine dinucleotide phosphate (nicotinamide adenine dinucleotide
Phosphate abbreviation), once referred to as codehydrogenase II (TPN) or secondary dehydrogenase II or oxidized coenzyme Ⅱ.It is one
Kind of coenzyme, be niacin hydroxyacyl amine adenine-dinucleotide with a phosphoric acid molecules with the substance in conjunction with ester bond, be widely present living nature.
The similar NAD such as chemical property, absorption spectrum, redox form.
NADP can carry out the synthesis of enzyme by the ATP phosphorylation of NAD+ in the prior art.
Currently, there are many ways to preparation nicotinamide adenine dinucleotide (NAD), including chemical synthesis, biological enzyme are urged
Change and saccharomycetes to make fermentation extraction etc..Chemical synthesis by chemical industry it is intrinsic seriously polluted, condition is harsh, and yield is not in addition
Height, synthesis cost are higher.For the method that saccharomycetes to make fermentation extracts, since nicotinamide adenine dinucleotide (NAD) is in yeast
Content in bacterium is too low, therefore this method has significant limitation on industrial production scale.Biological enzyme is because it is solid
Some high efficiency, it is environmentally protective the advantages that be increasingly becoming the main stream approach of industrially prepared nicotinamide adenine dinucleotide (NAD).
It mainly includes following key step that biological enzyme, which prepares nicotinamide adenine dinucleotide (NAD):
Step 1: with niacinamide ribose (nicotinamide riboside, NR) for starting material, in niacinamide ribose
Under the catalysis of kinases (nicotinamide riboside kinase, NRK), nicotinamide mononucleotide is obtained
(nicotinamidemononucleotide,NMN)。
Step 2: nicotinamide mononucleotide (nicotinamidemononucleotide, NMN) is in nicotinamide mononucleotide
It is and another under the action of adenylase (nicotinamidemononucleotide adenylyltransferase, NMNAT)
The reaction of one precursor atriphos (ATP), generates nicotinamide adenine dinucleotide (NAD).
But above-mentioned nicotinamide adenine dinucleotide (NAD) biological enzyme preparation method is related to multistep reaction, and yield
It is not high.
Summary of the invention
It is an object of the invention to solve to prepare nicotinamide adenine dinucleotide using biological enzyme in the prior art
When, the not high problem of complicated for operation and yield；A kind of preparation method of the NADP coenzyme to solve the above problems is provided.
In order to achieve the above objectives, technical scheme is as follows:
A kind of preparation method of NADP coenzyme, comprising:
Step 1: obtaining nicotinamide adenine dinucleotide；
NADP is made after ATP phosphorylation in nicotinamide adenine dinucleotide by second step；
Wherein, the preparation process of nicotinamide adenine dinucleotide are as follows:
Starting material, niacinamide ribokinase, nicotinamide mononucleotide adenylase and atriphos are added simultaneously
Enter into reaction vessel reaction and generates nicotinamide adenine dinucleotide；The concentration of the starting material is 30~40g/L, nicotinoyl
The ratio that amine ribokinase and nicotinamide mononucleotide adenylase are added is 2~3 ︰ 1~2；The niacinamide ribokinase
Additional amount be starting material weight 3~5%；The molar ratio of the atriphos and starting material is 2.1~2.3 ︰ 1.
It is reacted in the present invention by the way that substrate etc. to be all once added in reaction vessel, the operation is more convenient.Existing skill
It has been generally acknowledged that niacinamide ribokinase, nicotinamide mononucleotide adenylase are added to starting material niacinamide core simultaneously in art
It when in sugar, is easy to cause the product of generation that can't only there was only nicotinamide adenine dinucleotide, also generates that a large amount of other are miscellaneous
Matter, and impurity content is higher, yield is lower, and the separating-purifying time is more complex, is not suitable for industrialized production.
The present invention can effectively promote the product generated to be by the optimal control of additional proportion and additional amount between each substance
Nicotinamide adenine dinucleotide, and then NADP is prepared by nicotinamide adenine dinucleotide, yield of the invention is opposite
It is higher, it is suitable for industrialized production.
Further, reaction temperature is 20~25 DEG C in the reaction vessel, shake culture, and incubation time is 4~10h.Institute
The concentration for stating starting material is 32~35g/L, the ratio that niacinamide ribokinase and nicotinamide mononucleotide adenylase are added
Example is 3 ︰ 2；The additional amount of the niacinamide ribokinase is the 4% of starting material weight.The atriphos and starting bottom
The molar ratio of object is 2.2 ︰ 1.
By the optimal setting of above-mentioned condition, yield effectively can be increased to 60%, also, the time is reduced within 10h,
Production efficiency is higher, yield is also very significant.
Compared with prior art, the present invention have the following advantages that and the utility model has the advantages that
1, the present invention can effectively promote the product generated by the optimal control of additional proportion and additional amount between each substance
For nicotinamide adenine dinucleotide, and then NADP is generated, the present invention effectively promotes yield is opposite to mention while simplifying operation
Height is suitable for industrialized production；
2, the yield for the raw material nicotinamide adenine dinucleotide for producing NADP effectively can be increased to 60% by the present invention, and
And the time is reduced within 10h, production efficiency is higher, yield is also very significant.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, embodiments of the present invention are not limited thereto.
Embodiment 1
A kind of preparation method of NADP coenzyme, comprising:
Step 1: obtaining nicotinamide adenine dinucleotide；
Using niacinamide ribose as starting material, while by starting material, niacinamide ribokinase, nicotinamide mononucleotide gland
Glycosides transferase and atriphos, which are added in reaction vessel, to react, and reaction process is as follows:
Firstly, starting material niacinamide ribose reacts to obtain under the catalysis of niacinamide ribokinase, with atriphos
Nicotinamide mononucleotide；Secondly, nicotinamide mononucleotide is under the action of nicotinamide mononucleotide adenylase, with another three
Adenosine phosphate reaction, generates nicotinamide adenine dinucleotide.
In the present embodiment, reaction temperature is 20~25 DEG C in the reaction vessel, shake culture, incubation time 10h.Institute
The concentration for stating starting material is 32~35g/L, the ratio that niacinamide ribokinase and nicotinamide mononucleotide adenylase are added
Example is 3 ︰ 2；The additional amount of the niacinamide ribokinase is the 4% of starting material weight.The atriphos and starting bottom
The molar ratio of object is 2.2 ︰ 1.
It is learnt by detection, the yield of product nicotinamide adenine-dinucleotide is 62% in the present embodiment.
NADP is made after ATP phosphorylation in nicotinamide adenine dinucleotide by second step.Cigarette is used in the present embodiment
The process that NADP is made in amide adenine-dinucleotide after ATP phosphorylation is the prior art, therefore in the present embodiment no longer
It repeats.
Embodiment 2
The present embodiment the difference from embodiment 1 is that, part parameter setting is different in the present embodiment, is specifically provided that
The concentration of the starting material is 38g/L, and niacinamide ribokinase and nicotinamide mononucleotide adenylase add
The ratio entered is 3 ︰ 1；The additional amount of the niacinamide ribokinase is the 5% of starting material weight；
The molar ratio of the atriphos and starting material is 2.2 ︰ 1.
Yield in the present embodiment is 56%.
Embodiment 3
The present embodiment the difference from embodiment 1 is that, part parameter setting is different in the present embodiment, is specifically provided that
The concentration of the starting material is 20g/L, and niacinamide ribokinase and nicotinamide mononucleotide adenylase add
The ratio entered is 1 ︰ 1；The additional amount of the niacinamide ribokinase is the 6% of starting material weight；
The molar ratio of the atriphos and starting material is 2 ︰ 1.
Yield in the present embodiment is 47%.
Above-described embodiment is merely a preferred embodiment of the present invention, and it is not intended to limit the protection scope of the present invention, as long as using
Design principle of the invention, and the non-creative variation worked and made is carried out on this basis, it should belong to of the invention
Within protection scope.
Claims (4)
1. a kind of preparation method of NADP coenzyme, comprising:
Step 1: obtaining nicotinamide adenine dinucleotide；
NADP is made after ATP phosphorylation in nicotinamide adenine dinucleotide by second step；It is characterized in that,
The preparation process of nicotinamide adenine dinucleotide are as follows:
Starting material, niacinamide ribokinase, nicotinamide mononucleotide adenylase and atriphos are added to simultaneously
Reaction generates nicotinamide adenine dinucleotide in reaction vessel；The concentration of the starting material is 30~40g/L, niacinamide core
The ratio that sugared kinases and nicotinamide mononucleotide adenylase are added is 2~3 ︰ 1~2；The niacinamide ribokinase adds
Enter 3~5% that amount is starting material weight；The molar ratio of the atriphos and starting material is 2.1~2.3 ︰ 1.
2. a kind of preparation method of NADP coenzyme according to claim 1, which is characterized in that reacted in the reaction vessel
Temperature is 20~25 DEG C, shake culture, and incubation time is 4~10h.
3. a kind of preparation method of NADP coenzyme according to claim 2, which is characterized in that the concentration of the starting material
For 32~35g/L, the ratio that niacinamide ribokinase and nicotinamide mononucleotide adenylase are added is 3 ︰ 2；The nicotinoyl
The additional amount of amine ribokinase is the 4% of starting material weight.
4. a kind of preparation method of NADP coenzyme according to claim 1, which is characterized in that the atriphos with rise
The molar ratio of beginning substrate is 2.2 ︰ 1.
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| CN201811016614.3A CN108998484A (en) | 2018-09-03 | 2018-09-03 | A kind of preparation method of NADP coenzyme |
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| CN201811016614.3A CN108998484A (en) | 2018-09-03 | 2018-09-03 | A kind of preparation method of NADP coenzyme |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110643587A (en) * | 2019-10-29 | 2020-01-03 | 杭州唯泰生物药业有限公司 | Method for preparing nicotinamide adenine dinucleotide phosphate by enzyme method |
| CN111424064A (en) * | 2020-04-20 | 2020-07-17 | 比瑞博生物科技(北京)有限公司 | High-purity NMN preparation process based on enzyme method |
| CN111635917A (en) * | 2020-06-11 | 2020-09-08 | 辽宁天华生物药业有限公司 | Preparation method of beta-nicotinamide ribodinucleotide |
| WO2020258111A1 (en) * | 2019-06-27 | 2020-12-30 | 邦泰合盛生物科技（深圳）有限公司 | Enzymatic method for industrial production of nad |
| CN112437811A (en) * | 2019-06-27 | 2021-03-02 | 邦泰生物工程(深圳)有限公司 | Recombinant NAD synthetase, gene and application thereof |
-
2018
- 2018-09-03 CN CN201811016614.3A patent/CN108998484A/en not_active Withdrawn
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020258111A1 (en) * | 2019-06-27 | 2020-12-30 | 邦泰合盛生物科技（深圳）有限公司 | Enzymatic method for industrial production of nad |
| CN112437813A (en) * | 2019-06-27 | 2021-03-02 | 邦泰生物工程(深圳)有限公司 | Method for industrially producing NAD (nicotinamide adenine dinucleotide) by enzyme method |
| CN112437811A (en) * | 2019-06-27 | 2021-03-02 | 邦泰生物工程(深圳)有限公司 | Recombinant NAD synthetase, gene and application thereof |
| CN112437813B (en) * | 2019-06-27 | 2023-03-03 | 邦泰生物工程(深圳)有限公司 | Method for industrially producing NAD (nicotinamide adenine dinucleotide) by enzyme method |
| CN112437811B (en) * | 2019-06-27 | 2023-05-05 | 邦泰生物工程(深圳)有限公司 | Recombinant NAD synthetase, gene and application thereof |
| CN110643587A (en) * | 2019-10-29 | 2020-01-03 | 杭州唯泰生物药业有限公司 | Method for preparing nicotinamide adenine dinucleotide phosphate by enzyme method |
| CN111424064A (en) * | 2020-04-20 | 2020-07-17 | 比瑞博生物科技(北京)有限公司 | High-purity NMN preparation process based on enzyme method |
| CN111635917A (en) * | 2020-06-11 | 2020-09-08 | 辽宁天华生物药业有限公司 | Preparation method of beta-nicotinamide ribodinucleotide |
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Application publication date: 20181214 |