CN1108660A - Technology for production of adenosine triphospharic acid (ATP) by using adenosine (AR) as raw material - Google Patents
Technology for production of adenosine triphospharic acid (ATP) by using adenosine (AR) as raw material Download PDFInfo
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- CN1108660A CN1108660A CN 95100148 CN95100148A CN1108660A CN 1108660 A CN1108660 A CN 1108660A CN 95100148 CN95100148 CN 95100148 CN 95100148 A CN95100148 A CN 95100148A CN 1108660 A CN1108660 A CN 1108660A
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Abstract
The tecnology for production of adenosine triphosph (ATP) by using adenosin (AR)as raw material is that high-concentration (above 2%) conversion method using AR as bottom material and alcoholic-deposition desalting technology are adopted to make the adsorption ratio of resin increse from original 5-13g ATP/100g resin to 18-22g ATP/100g resin, the content of upper-column liquid, cleaning mixture and eluent and the temp. under 15 deg. C can be controlled to make the cost reduce to 1500 Yuan/Kg. The content of said product is over ministry standard by above 11%.
Description
The present invention relates to a kind of biochemical pharmacy novel process, particularly a kind of is the novel process of raw material production adenosine triphosphate (ATP) with adenosine (AR).
At the late nineteen seventies early eighties, China mainly contains four tame production of units adenosine triphosphates (ATP), it is respectively Jiangmen sugar chemical factory, grape sugar refinery, Shanghai, North China Pharmaceutical Factory, Jiamusi biochemical-pharmaceutical factory, all be to be raw material with adenylic acid (AMP), by the relevant enzyme in make the transition yeast or the cereuisiae fermentum is to produce adenosine triphosphate (ATP), but, adopt above processing technology routine to produce ATP, cost is very high, reach 3000～4000 yuan/kg, and import ATP to advance the price of main China market only be about 2200 yuan/kg, closely like this make these the whole stopping productions of China ATP factory, it is in paralyzed state substantially that China produces ATP industry.
At above situation, units such as Huadong Chemical College, the biochemical institute in Shanghai, the organic institute in Shanghai, the late nineteen eighties begin one's study with adenosine (AR) be the technical matters of raw material production ATP medical material, they at first carry out pilot scale at Qidong, Jiangsu Province county plant nucleotide, but owing to the Technology reason, quality product does not still reach Ministry of Health's standard, and production cost still is higher than the imported product price, can't formally produce, it is as follows to study carefully its major cause:
1, fermentation conversion concentration is low
Difficulty is extracted in low cause (1) of fermentation concentration, makes the extraction cost height; (2) the fermentation equipment utilization ratio is low, the production stable cost is big; (3) labour productivity is low, and production cost increases.
2, prescription is unreasonable
This is because the subsidiary material use of fermenting is too high, increases materials consumption, and cost is improved, and more the order extraction is difficult, and productive rate and quality product descend.
3, zymotechnique is unreasonable
Mainly be that pH value does not reach and makes AR be converted into the ATP optimum value, thereby make the product recovery rate low, make the production cost height.
4, extraction process is unreasonable
Mainly be that upper prop ion exchange resin adsorption rate in the upper prop purifying technique is low, be generally 5～13gATP/100g resin, and make recovery and plant factor is low, product content is low, downgrade.
The objective of the invention is the improvement that exists as above problem to propose at the technology that with AR is raw material production ATP.
Its reaction process is as follows:
Adopt following technical process:
AR solution+clear enzyme solution carries out ATP fermentation → centrifugation → alcohol precipitation → throw out Hui Rong → dilution upper prop purification → depyrogenation → crystallization → vacuum-drying → ATP.
1, adopt high density AR conversion method, concentration is more than 2%.
(1) prescription (seeing Table)
Example:
| Name of material | Specification | Quantity | Remarks |
| AR | More than 90% | 1.5～2.5kg | Water is made into 20% solution |
| Saccharase liquid | 38～451 |
(2) processing parameter:
PH:6.3～6.7 temperature: 32～35 ℃ of times: 4～6 hours
Transformation efficiency: more than 90%;
End point determination: DEAE-cellulose sheet layer watchman's clapper follow-up investigations.
2, adjust the ATP zymotechnique, make it be more suitable for AR and be converted into ATP, obviously improved the yield of product, the transformation efficiency that makes AR is more than 90%.
3, adopt the alcohol precipitation desalinating process, the adsorption rate that makes resin is brought up to 18～22gATP/100g resin by original 5～13gATP/100g resin.
(1) processing parameter:
Fermentation separates clear liquid: alcohol consumption=1: 4.0～5.0＜weight 〉
Stir speed (S.S.): 40～50 rev/mins
Churning time: 30～40 minutes
The quiescent setting time: 4～8 hours
Keep fermentation to separate the pH value of clear liquid: 2.0～2.2
ATP content g/l:35～60 of fermentation parting liquid;
4, the temperature of the upper prop liquid in the control ATP upper prop purifying technique process, washings, elutriant improves recovery and quality product that the ATP upper prop is purified below 15 ℃.
(1) upper prop processing parameter
Upper column quantity: 18～22gATP/100g201 * 7 ion exchange resin, 100～200 orders;
Upper prop liquid ATP content 1～2%;
Upper prop liquid pH value: 3.2～3.8;
Upper prop liquid temp: below 15 ℃;
Last column flow rate: 20～30ml/min.kg resin;
(2) washing process parameter
Washings composition: 0.01MNaCl or PH2.0 solution;
Washing flow velocity: 40～50ml/min.kg resin;
Washing time: 30～40 hours;
Wash temperature: below 15 ℃.
The washing terminal point is with the follow-up investigations of DEAE fiber thin flaggy watchman's clapper, confirms behind the no ATP 3～8 hours.
(3) elutriant composition: 1MNaCl-PH3.8HCl solution (pyrogen-free);
Eluent flow rate: early stage 10～15ml/min.kg resin;
Collection phase 3～5ml/min.kg resin.
Collection method: denseer with the alcohol precipitation inspection, ATP content is more than 70g/l.
By above improvement, make the ATP cost drop to 1500 yuan/kg by 3000～4000 yuan original/kg, product content surpasses Ministry of Health's standard more than 11%, has produced remarkable economic efficiency and social benefit.
Describe the present invention below in detail:
1, the reaction process with AR production ATP is as follows:
Its enzyme extraction technical process is as follows:
Separate → disappear enzyme liquid
The technology of producing ATP is as follows:
AR solution+clear enzyme solution carries out ATP fermentation → centrifugation → alcohol precipitation → throw out Hui Rong → dilution upper prop purification → depyrogenation → crystallization → vacuum-drying → ATP.
The optimised process of producing ATP is as follows:
AR solution+clear enzyme solution carries out ATP fermentation → termination reaction → centrifugation → clear liquid → alcohol precipitation → throw out → Hui Rong → freeze desalination → diatomite processing → filtration (press filtration) → clear liquid → dilution upper prop purification → washing → wash-out → decolorizing resin decolouring → diatomite processing → gac depyrogenation processing → filtration (G5 funnel) → crystallization → crystallisate → absolute ethanol washing → anhydrous diethyl ether washing → vacuum-drying → ATP.
2, technology condition
(1) beer yeast slurry or beer yeast cake:
Beer yeast slurry is the yeast with greater activity collected in the beer production, and amount of yeast in the beer yeast slurry (weight percent) is 16-18%, amount of yeast in the yeast cake dry-matter (weight percent) 23-26%.
(2) the refrigerating process condition of cereuisiae fermentum:
1) freezing temp :-15 ℃-25 ℃
2) freezing time: 7 days-15 days
3) quick-frozen is frozen the time: in 8 hours
(3) the enzymatic process condition is put forward in fermentation:
1) prescription
Name of material specification quantity remarks
Available 1%-2% before beer yeast slurry 16-18% 20～50kg adds
Bromogeramine was handled 30 minutes
In phosphoric acid 85% (industry) 2～5kg KOH and the phosphoric acid and back is used
NaOH transfers PH6.6-6.7
The fixed molten 15l of KOH CP 1～3kg
MgCl
2CP 0.1～2kg is made into 25% and adds
Sucrose eats 3～6kg
Total volume 50-60l
2) control pH value: 6.3-6.7
3) temperature: 32-35 ℃
4) time: 2-2.5 hour
5) the residual body of yeast separates: centrifugation
(4) AR is converted into the processing condition of ATP:
1) prescription
Name of material specification quantity remarks
AR 90% 1.0-3.0kg water distribution becomes 20%
The above-mentioned parting liquid 30-45l of saccharase liquid
Sucrose eats 0.5-1.0kg
Phosphoric acid industry 0.2-0.5kg
Add in the enzyme liquid for preparing again after preparing.
2) processing condition:
PH：6.3-6.7
Temperature: 32-35 ℃
Time: 4-7 hour
More than the transformation efficiency %:90%
End point determination: DEAE-cellulose sheet chromatography follow-up investigations ATP transformation efficiency is more than 90%.
(5) termination reaction:
Termination reaction agent: technical hydrochloric acid
Stop pH value: PH1.9-2.0
(6) centrifugation removes albumen:
Test manufacture: filtration medium: nylon cloth
Separating machine: 33-800 link-suspended basket centrifuge
Pilot scale: tubular-bowl centrifuge
Centrifugation clear liquid standard: not muddy, no suspended substance.
(7) alcohol precipitation:
ATP clear liquid pH value: 1.9-2.1
Alcohol consumption: 4.5-5.0 doubly
Mixing speed: 40-50 rev/min
Churning time: 30-60 minute
Quiescent setting time: 4-8 hour
Fermentation separates clear liquid ATP content g/l:45-70g/l
(8) ATP throw out Hui Rong:
Solvent: distilled water
ATP concentration: 200g/l-220g/l
(9) freeze desalination:
Temperature :-5--15 ℃
The desalination standard: the icing back suction filtration that salts out is removed salt.
(10) diatomite is handled:
ATP concentration: 5%
Diatomite consumption: 1%
Temperature: below 25 ℃
Time: stirred 30 minutes
(11) press filtration or suction filtration:
Press filtration sterile filtration machine
Medium filter paper
The suction filtration vacuumfilter
(12) upper prop is purified:
Resin model: 201 * 8(201 * 7) 100-200 purpose strong basic type anion-exchange resin.
Resin demand: 18-20gATP/100g resin
Last column liquid concentration: ATP content 10g/l
Upper prop liquid PH:2.8～3.5;
Last column temperature: below 20 ℃;
Last column flow rate: 20～30ml/min.kg resin;
The upper prop terminal point: the inspection of DEAE-cellulose sheet chromatography has the ATP spot
Washing composition: 0.01MNaCl-0.01HCl solution (pyrogen-free) PH2.0
Wash temperature: below 18 ℃.
Washing flow velocity: 40～50ml/min.kg resin;
The washing terminal point: the inspection of DEAE-cellulose sheet chromatography does not have behind the ADP 3-8 hour
Eluent: 1MNaCl-PH3.8HCl solution (pyrogen-free);
Elution flow rate: in earlier stage: 10～15ml/min.kg resin (time: 30-40 minute)
The collection phase: 3～5ml/min.kg resin
Collection method: with the denseer ATP content of alcohol precipitation inspection more than 70g/l
Collect terminal point: ATP content is below 10g/l
(13) refining:
Decolouring is handled: flow through the decolorizing resin decolouring with elutriant
Flow velocity: 50-100ml/min.kg resin
Transmittance: more than 99%
Diatomite is handled:
Diatomite consumption: 1% of destainer
Treatment time: stirred 30-40 minute
Suction filtration vacuum tightness :-0.1MPa
Gac removes thermal source:
Pharmaceutical powder activated carbon dosage: the first time 0.3%, the second time 0.15%
Suction filtration vacuum tightness :-0.1MPa
Positively charged ion removing heavy metals: with suction filtration behind the positively charged ion accent PH2.5.
The G5 funnel filters:
Suction filtration vacuum tightness :-0.1MPa
Annotate: above refining whole process should prevent that thermal source from polluting.
(14) ATP crystallization:
ATP clear liquid pH value: 2.5
Alcohol consumption: 2.5 times
Temperature: below 25 ℃
Time: 10-16 hour
(15) absolute ethanol washing crystallisate:
Dehydrated alcohol consumption: 0.5 times of throw out
Vacuum filtration is anhydrous to be flowed down
(16) anhydrous diethyl ether washing:
Anhydrous diethyl ether consumption: 0.5 times of throw out
Vacuum filtration is anhydrous to be flowed down, and crystallisate does not have the thickness sense, loose good dispersion.
(17) P
2O
5Vacuum-drying:
Vacuum tightness :-0.1MPa
Temperature: below 45 ℃
Finished product water content: below 7%
Two, test manufacture result
1, main economic and technical norms result (tabulation as follows)
Table-2 test manufacture ATP test-results
Table-3 ATP test products quality results
Table-4 ATP test manufacture is production cost as a result
Continuous table-4
Claims (8)
1, a kind of is the novel process of raw material production adenosine triphosphate (ATP) with adenosine (AR), and its technical process is as follows:
AR solution+clear enzyme solution carries out ATP fermentation → centrifugation → alcohol precipitation → throw out Hui Rong → dilution upper prop purification → depyrogenation → crystallization → vacuum-drying → ATP.
2, a kind of usefulness adenosine according to claim 1 (AR) is the novel process of raw material production adenosine triphosphate (ATP), and its technical process is preferably as follows:
AR solution+clear enzyme solution carries out ATP fermentation → termination reaction → centrifugation → clear liquid → alcohol precipitation → throw out → Hui Rong → freeze desalination → diatomite processing → filtration (press filtration) → clear liquid → dilution upper prop purification → washing → wash-out → decolorizing resin decolouring → diatomite processing → gac depyrogenation processing → filtration (G5 funnel) → crystallization → crystallisate → absolute ethanol washing → anhydrous diethyl ether washing → vacuum-drying → ATP.
3, a kind of usefulness adenosine according to claim 1 (AR) is the novel process of raw material production adenosine triphosphate (ATP), and the reparation technology of its clear enzyme solution is as follows:
4, a kind of usefulness adenosine according to claim 1 (AR) is the novel process of raw material production adenosine triphosphate (ATP), it is characterized in that adopting the precipitation desalinating process, makes the adsorption rate of resin bring up to 18～22gATP/100g resin, and its processing parameter is as follows:
Separation clear liquid after the fermentation: alcohol consumption=1: 4.0～5.0＜weight 〉
Stirring velocity: 40～50 rev/mins
Churning time: 30～40 minutes
The quiescent setting time: 4～8 hours
The pH value of the separation clear liquid after keeping fermenting is: 2.0～2.2
The ATP content of fermentation parting liquid is: 35～60g/l
5, a kind of usefulness adenosine according to claim 1 (AR) is the novel process of raw material production adenosine triphosphate (ATP), the temperature that it is characterized in that controlling upper prop liquid in the ATP upper prop purifying technique process, washings, elutriant is below 15 ℃, so that improve recovery and quality product that the ATP upper prop is purified, its
(1) the upper prop processing parameter is:
Upper column quantity: 18～22gATP/100g201 * 7 ion exchange resin, 100～200 orders;
Upper prop liquid: ATP content 1～2%;
Upper prop liquid pH value: 3.2～3.8;
Upper prop liquid temp: below 15 ℃;
Last column flow rate: 20～30ml/min.kg resin;
(2) the washing process parameter is:
Washings composition: 0.01MNaCl-PH2.0HCl solution;
Washing flow velocity: 40～50ml/min.kg resin;
Washing time: 30～40 hours;
Wash temperature: below 15 ℃.
The washing terminal point is with the follow-up investigations of DEAE fiber thin flaggy watchman's clapper, confirms behind the no ATP 3～8 hours.
(3) elution processes parameter:
Elutriant composition: 1MNaCl-PH3.8HCl solution (pyrogen-free);
Eluent flow rate: early stage 10～15ml/min.kg resin;
Collection phase 3～5ml/min.kg resin.
6, a kind of with adenosine (AR) be the novel process of raw material production adenosine triphosphate (ATP), adenosine (AR) water is made into the 20%(weight ratio) solution after the adding clear enzyme solution in, make the content of adenosine (AR) in whole solution greater than the 2%(weight ratio).
7, a kind of usefulness adenosine according to claim 6 (AR) is the novel process of raw material production adenosine triphosphate (ATP), the best according to the form below prescription of the content of adenosine in whole solution:
8, a kind of usefulness adenosine according to claim 6 (AR) is the novel process of raw material production adenosine triphosphate (ATP), and the processing parameter when adenosine (AR) ferments in clear enzyme solution is:
PH value: 6.3～6.7 temperature: 32～35 ℃
Time: 4～6 hours
End point determination: DEAE-cellulose sheet layer watchman's clapper follow-up investigations ATP transformation efficiency is more than 90%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95100148 CN1108660A (en) | 1995-01-17 | 1995-01-17 | Technology for production of adenosine triphospharic acid (ATP) by using adenosine (AR) as raw material |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95100148 CN1108660A (en) | 1995-01-17 | 1995-01-17 | Technology for production of adenosine triphospharic acid (ATP) by using adenosine (AR) as raw material |
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| CN1108660A true CN1108660A (en) | 1995-09-20 |
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|---|---|---|---|
| CN 95100148 Pending CN1108660A (en) | 1995-01-17 | 1995-01-17 | Technology for production of adenosine triphospharic acid (ATP) by using adenosine (AR) as raw material |
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|---|---|---|---|---|
| CN102584923A (en) * | 2012-01-30 | 2012-07-18 | 烟台大学 | Method for extracting compounds from Chondrus ocelltus |
| CN104762347A (en) * | 2015-03-16 | 2015-07-08 | 浙江工商大学 | Production method of adenosine triphosphate (ATP) |
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| CN104861022A (en) * | 2015-05-28 | 2015-08-26 | 广西浦北制药厂 | Preparation technology of disodium adenosine triphosphate capable of reducing energy consumption |
| CN104878058A (en) * | 2015-05-28 | 2015-09-02 | 广西浦北制药厂 | Disodium adenosine triphosphate production method |
| CN104894192A (en) * | 2015-05-28 | 2015-09-09 | 广西浦北制药厂 | Preparation method for adenosine disodium triphosphate |
| CN104928334A (en) * | 2015-05-28 | 2015-09-23 | 广西浦北制药厂 | Processing technique of disodium adenosine triphosphate |
| CN109851650A (en) * | 2018-12-19 | 2019-06-07 | 新疆阜丰生物科技有限公司 | A method of extracting adenosine from fermentation liquid |
-
1995
- 1995-01-17 CN CN 95100148 patent/CN1108660A/en active Pending
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| CN102584923B (en) * | 2012-01-30 | 2014-01-15 | 烟台大学 | Method for extracting compounds from Chondrus ocelltus |
| CN104762347A (en) * | 2015-03-16 | 2015-07-08 | 浙江工商大学 | Production method of adenosine triphosphate (ATP) |
| CN104762347B (en) * | 2015-03-16 | 2018-05-08 | 浙江工商大学 | A kind of production method of atriphos (ATP) |
| CN104862357A (en) * | 2015-05-28 | 2015-08-26 | 广西浦北制药厂 | Production method of adenosine disodium triphosphate |
| CN104862359A (en) * | 2015-05-28 | 2015-08-26 | 广西浦北制药厂 | Production method of adenosine disodium triphosphate |
| CN104846038A (en) * | 2015-05-28 | 2015-08-19 | 广西浦北制药厂 | Process for preparing trinosin |
| CN104846042A (en) * | 2015-05-28 | 2015-08-19 | 广西浦北制药厂 | Energy-saving production method of adenosine disodium triphosphate |
| CN104846040A (en) * | 2015-05-28 | 2015-08-19 | 广西浦北制药厂 | Method for processing trinosin |
| CN104846041A (en) * | 2015-05-28 | 2015-08-19 | 广西浦北制药厂 | Method for preparing trinosin |
| CN104830929A (en) * | 2015-05-28 | 2015-08-12 | 广西浦北制药厂 | Processing method of trinosin |
| CN104862358A (en) * | 2015-05-28 | 2015-08-26 | 广西浦北制药厂 | Production method of improved adenosine disodium triphosphate |
| CN104862360A (en) * | 2015-05-28 | 2015-08-26 | 广西浦北制药厂 | Preparation method of disodium adenosine triphosphate capable of reducing energy consumption |
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| CN104861022A (en) * | 2015-05-28 | 2015-08-26 | 广西浦北制药厂 | Preparation technology of disodium adenosine triphosphate capable of reducing energy consumption |
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| CN104894192A (en) * | 2015-05-28 | 2015-09-09 | 广西浦北制药厂 | Preparation method for adenosine disodium triphosphate |
| CN104928334A (en) * | 2015-05-28 | 2015-09-23 | 广西浦北制药厂 | Processing technique of disodium adenosine triphosphate |
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