CN101328489A - Method for yeast cell to biosynthesizing 1,6- fructose sodium diphosphate - Google Patents
Method for yeast cell to biosynthesizing 1,6- fructose sodium diphosphate Download PDFInfo
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- 210000005253 yeast cell Anatomy 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 23
- 230000003570 biosynthesizing effect Effects 0.000 title claims description 9
- 238000006243 chemical reaction Methods 0.000 claims abstract description 41
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 20
- 210000004027 cell Anatomy 0.000 claims abstract description 20
- 239000011734 sodium Substances 0.000 claims abstract description 19
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 19
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 17
- 239000008103 glucose Substances 0.000 claims abstract description 17
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 230000001954 sterilising effect Effects 0.000 claims abstract description 5
- 239000002253 acid Substances 0.000 claims description 18
- 229940045641 monobasic sodium phosphate Drugs 0.000 claims description 13
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 12
- 229920000053 polysorbate 80 Polymers 0.000 claims description 12
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
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- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 abstract description 8
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- CEHGLSKJDFICTB-PWVOXRODSA-K D-Fructose 1,6-bisphosphate trisodium salt Chemical compound [Na+].[Na+].[Na+].[O-]P(=O)([O-])OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)COP(O)([O-])=O CEHGLSKJDFICTB-PWVOXRODSA-K 0.000 description 2
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- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
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- 208000028399 Critical Illness Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000003793 Fructokinases Human genes 0.000 description 1
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- 241000193385 Geobacillus stearothermophilus Species 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 1
- 102000018120 Recombinases Human genes 0.000 description 1
- 108010091086 Recombinases Proteins 0.000 description 1
- CWHJIJJSDGEHNS-MYLFLSLOSA-N Senegenin Chemical compound C1[C@H](O)[C@H](O)[C@@](C)(C(O)=O)[C@@H]2CC[C@@]3(C)C(CC[C@]4(CCC(C[C@H]44)(C)C)C(O)=O)=C4[C@@H](CCl)C[C@@H]3[C@]21C CWHJIJJSDGEHNS-MYLFLSLOSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
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- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000001177 diphosphate Substances 0.000 description 1
- 235000011180 diphosphates Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
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- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
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- 235000013580 sausages Nutrition 0.000 description 1
- HBYYJIPCYANMBX-BAOOBMCLSA-M sodium;[(3s,4r,5r)-3,4,5-trihydroxy-2-oxo-6-phosphonooxyhexyl] hydrogen phosphate Chemical compound [Na+].OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)COP(O)([O-])=O HBYYJIPCYANMBX-BAOOBMCLSA-M 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for biologically synthesizing fructose 1,6 diphosphate sodium of a yeast cell, comprising the following steps that: based on 100mL of sterilizing reaction fluid, 1 to 5 grams of thalli obtained by pre-culture process is added to react for 4 to 10 hours at a temperature of between 25 and 30 DEG C and at a speed of 100 to 200r/min, trichloroacetic acid is added to stop the reaction, the clear solution of the fructose 1,6 diphosphate sodium is obtained by centrifugalization; the thalli is cells which are obtained by pre-culturing for 12 to 36 hours, freezing and centrifuging of Saccharomyces cerevisiae; the pH value of the reaction fluid is between 5.0 and 9.0, the reaction fluid comprises the following compositions in weight percentage: 2 to 12 percent of glucose, 2 to 15 percent of sodium dihydrogen phosphate, and the balance being water. The fructose 1,6 diphosphate sodium synthesized by the method has simple process for culturing the yeast cells, shortens the time, and has simple converting system, easy operation and high FDP sodium converting efficiency.
Description
Technical field
The invention belongs to biotechnology and microbial fermentation field, relate in particular to a kind of yeast cell biosynthesizing 1, the method for 6-hexose diphosphate sodium.
Background technology
1,6-hexose diphosphate [Fructose-1,6-Diphosphate are called for short FDP], molecular formula is C
6H
14O
12P
2, often the form with salt such as sodium, calcium, zinc exists.Be glycometabolic intermediate product, water-soluble, be that the human life activity is necessary, in cellular process, work the vital role of joint agent, biological catalyst and the cell tonic of withering.
Since Harden in 1908 and Young found and illustrate the fundamental characteristics of 1,6 one hexose diphosphate, many scientists were to the preparation of 1,6 one hexose diphosphate and should be used as research.People such as Nenfeng tie up to the laboratory in nineteen forty-two with the enzyme of fresh bread yeast or cereuisiae fermentum and have synthesized FDP, further confirm 1,6 one hexose diphosphates are as the argument of the intermediate in the carbohydrate metabolism process, and people such as Markov then lay the foundation for the application of FDP in medical science to pharmacology and the pharmacodynamic study that FDP carries out.Their FDP that discovers can act on cytolemma, regulate some enzymic activitys in the carbohydrate metabolism, especially activating phosphatase fructokinase (PFK), the poly-interior energy-rich phosphate root of cell that increases, produce a large amount of ATP, promote stream in the potassium ion, recover the cell polarized state, thereby be of value to the cellular energy metabolism under the states such as shock, anoxic, ischemic, damage, extracorporeal circulation and blood transfusion and the utilization of Liu's glucose, to promote cytothesis and to improve function.Clinically, FDP as activity and the recovery of regulating some enzymes in the carbohydrate metabolism, improve the molecular level medicine of cellular metabolism, be used for shock, Acute Myocardial Infarction, cardiac muscle direct-view operation, peripheral blood vessel illness and critical illness people and accept treatment of diseases and assisting therapy such as parenteral alimentation therapy.
The eighties, Italian Foscama company makes a breakthrough in fields such as the technology of preparing of FDP and clinical applications.First realized the suitability for industrialized production of FDP in the world, and has released the powder injection of 1,6 one fructose diphosphate sodium, obtains immense success, and its turnout is risen to 5,000,000 pins in 1991 by 20,000 pins in 1986.According to statistics, China's import volume in 1988 is 100,000 pins, increases sharply to more than 100 ten thousand pins, reaches more than 2,000,000 pins in 1993 in 1991.Domestic research to FDP is started late, and abroad accumulation mechanism and the production process of FDP is also lacked systematic study, does not see monograph so far.Except that the people such as Bisso of Foscama company have reported the preparation technology of FDP, do not see systematic article report in patent.
The 80-90 age finds 1,6-hexose diphosphate (Fructose-1,6-diphosphate) all multi-usages aspect Medicines and Health Product and plant growth regulating, being particularly useful for the receive much concern production of traditional F DP of medicine is to do the enzyme source with bread yeast or cereuisiae fermentum, obtain by glycolytic cycle, later useful fixed yeast cell, immobilization multienzyme system and bacillus stearothermophilus transforming glucose are the patent report of FDP, also have to adopt the reorganization cereuisiae fermentum to transform lactose be FDP and replace toluene to change the saturating property of yeast cell with CTAB, is the research etc. of carbon source biosynthesizing FDP with glucose.
Bio-transformation is to utilize plant isolated cells or organ, zooblast, microorganism and organoid thereof, and resolvase carries out the biochemical reaction of structural modification to exogenous compounds.Natural product is the human source of seeking the effective active composition always on the one hand; On the other hand, in the process of developing new drug, having with the natural product is lead compound, by chemistry or biological means, its structure is modified, and obtains the compound of the many novel structures of root, and then finds that toxicity is lower, the medicine of better efficacy.Microorganism is as a complete individuality, though its volume is little, also has the special enzyme system of a cover self.Because Microbial resources are abundant, of a great variety,, can supply the bacterial strain of screening a lot of simultaneously for substrate specially.Enzyme system with multiple different catalysiss carries out bio-transformation to active skull cap components, can produce the natural compounds storehouse of new associativity, pass through screening active ingredients again, can seek the natural radioactivity lead compound of new high-efficiency low-toxicity, or, huge social benefit and economic benefit have been obtained by can find the lead compound of a new generation to the heterogeneity structural changes in the active ingredient.The bio-transformation rule can also instruct the structural modification of new drug, obtains the medicine of new generation of efficient long-acting, is enough to illustrate the importance of bio-transformation to new drug development.
Now Bao Dao conversion synthetic method mostly is to adopt the fungal hyphae body as microorganism transformed, and this quasi-microorganism is cultivated loaded down with trivial details, time-consuming longer, and is unfavorable for the easy operated, and the efficient of biosynthesizing simultaneously is not high.
Summary of the invention
It is short to the invention provides a kind of cycle, the yeast cell biosynthesizing 1 that output is high, the method for 6-hexose diphosphate sodium.
A kind of yeast cell biosynthesizing 1, the method for 6-hexose diphosphate sodium comprises the steps:
Plum mountain yeast or yeast saccharomyces cerevisiae (trade name of yeast saccharomyces cerevisiae is a distillery yeast, the distillery yeast that can adopt Angel Yeast Co.,Ltd to produce) are seeded on the fermention medium, and fermention medium is that pol is 12 Berlin, and pH is 6.5 wort; What inoculation was adopted is the yeast starter liquid of logarithmic phase.Pre-12～the 36h that cultivates obtains fermented liquid in 25～30 ℃, 150r/min shaking table; At 4 ℃, under the 10000r/min condition fermented liquid frozen centrifugation 20～30min is obtained thalline.Preferred 18～24h of pre-incubated time, most preferably 24h.
The inventive method is not restricted to yeast saccharomyces cerevisiae, for plum mountain yeast, K word yeast etc. other (yeast is unicellular eukaryotic microorganisms.The form of yeast cell has sphere, oval, sausage shape, ellipse, lemon shape or rhizoma nelumbinis shape etc. usually, has typical eucaryotic cell structures, cell walls, cytolemma, nucleus, tenuigenin, vacuole, plastosome etc. are arranged, and what have also has a microbody) yeast is suitable for too.
After obtaining thalline with cultivation, in the reaction solution of 100mL sterilization, add 1～5g thalline, at 25～30 ℃, 100～200r/min conversion reaction, 4～10h, add 1mol/L Tricholroacetic Acid termination reaction; 4 ℃, the centrifugal 30min of 10000r/min collects supernatant liquor.
Contain 1 in the supernatant liquor, 6-hexose diphosphate sodium (FDP sodium) can adopt conventional extracting method to extract high-load FDP sodium from supernatant liquor, as the precipitator method, ion exchange adsorption etc.
For in the detected supernatant liquor 1,6-hexose diphosphate na concn can be handled supernatant liquor and survey light absorption value under wavelength 570nm by reported method.
Preferred as reaction conditions in the reaction solution of 100mL sterilization, adds the 3g thalline, at 30 ℃, 150r/min conversion reaction 6h, adds 1mol/L Tricholroacetic Acid termination reaction; 4 ℃, the centrifugal 30min of 10000r/min collects supernatant liquor.
The pH5.0 of described reaction solution～9.0, the mass percent of reaction solution consists of:
2～12% glucose;
2～15% SODIUM PHOSPHATE, MONOBASIC;
Surplus is a water.
The preferred pH5.5 of described reaction solution～8.9, the mass percent of reaction solution is formed preferred:
9%～11% glucose;
3%～7% SODIUM PHOSPHATE, MONOBASIC;
Surplus is a water.
Can improve the permeability of yeast cell as the preferred tensio-active agent Tween 80 (tween 80) that in reaction solution, adds reaction solution total mass 0.05～0.1%, improve the biosynthesizing amount of FDP.
When adding tensio-active agent Tween 80, the mass percent of reaction solution consists of:
9%～11% glucose;
3%～7% SODIUM PHOSPHATE, MONOBASIC;
0.05～0.1% tween 80;
Surplus is a water.
The inventive method, not only the yeast cell cultivation is simple, time-consuming shorter, and transformation system is simple, easy handling, FDP sodium combined coefficient height takes the tensio-active agent tween 80 to regulate and control and improve the combined coefficient of FDP sodium, has great application prospect.
Embodiment
Embodiment 1
Select the yeast saccharomyces cerevisiae bacterial classification for use, 30 ℃, (substratum: pol is the wort of 12 Berlin to 150r/min shaking table culturing cell, pH6.5, the bottled 100mL of 500mL triangle), the 24h collecting cell obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10000r/min.
Get the centrifugal yeast cell 1g that obtains, adding mass percent is 11% glucose, 5% SODIUM PHOSPHATE, MONOBASIC, the 30mL reaction solution of pH 7, invert point are 30 ℃, and rotating speed is 150r/min, after transforming 6h, with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), record in the fermented liquid 1 after treatment, 6-hexose diphosphate na concn is 4.79g/L.
Embodiment 2
Select the yeast saccharomyces cerevisiae bacterial classification for use, 30 ℃, the 150r/min shaking table is cultivated thalline, and (substratum: pol is the wort of 12 Berlin, pH6.5, the bottled 100mL of 500mL triangle), the 24h collecting cell obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10000r/min.
Get the centrifugal yeast cell 1g that obtains, adding mass percent is 8% glucose, 5% SODIUM PHOSPHATE, MONOBASIC, the 30mL reaction solution of pH8.4, invert point are 30 ℃, and rotating speed is 150r/min, after transforming 6h, with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), record in the fermented liquid 1 after treatment, 6-hexose diphosphate na concn is 3.9g/L.
Embodiment 3
Select the yeast saccharomyces cerevisiae bacterial classification for use, 30 ℃, (substratum: pol is the wort of 12 Berlin to 150r/min shaking table culturing cell, pH6.5, the bottled 100mL of 500mL triangle), the 24h collecting cell at J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10, obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of 000r/min.
Get the centrifugal yeast cell 1g that obtains, adding mass percent is 9.7% glucose, 5.8% SODIUM PHOSPHATE, MONOBASIC, the 30mL reaction solution of pH8.8, invert point are 30 ℃, and rotating speed is 150r/min, after transforming 6h, with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), record in the fermented liquid 1 after treatment, 6-hexose diphosphate na concn is 5.07g/L.
Embodiment 4
Select the yeast saccharomyces cerevisiae bacterial classification for use, 30 ℃, (substratum: pol is the wort of 12 Berlin to 150r/min shaking table culturing cell, pH6.5, the bottled 100mL of 500mL triangle), the 24h collecting cell obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10000r/min.
Get the centrifugal yeast cell 1g that obtains, adding mass percent is 9.7% glucose, 5.8% SODIUM PHOSPHATE, MONOBASIC, add tween 80 0.08%, the 30mL reaction solution of pH8.8, invert point is 30 ℃, rotating speed is 150r/min, after transforming 6h,, record in the fermented liquid 1 after treatment with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), 6-hexose diphosphate na concn is 5.67g/L, and the FDP sodium resultant quantity that does not add tween 80 in the same transformation system is 5.02g/L, 1, and the system that do not add tween 80 of 6-hexose diphosphate na concn under the similarity condition.
Embodiment 5
Select the yeast saccharomyces cerevisiae bacterial classification for use, 30 ℃, (substratum: pol is the wort of 12 Berlin to 150r/min shaking table culturing cell, pH6.5, the bottled 100mL of 500mL triangle), collecting cell behind the growth 24h obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10000r/min.
Get the centrifugal yeast cell 100g that obtains, adding mass percent is 9.7% glucose, 5.8% SODIUM PHOSPHATE, MONOBASIC, the 3000mL reaction solution of pH8.8 (5L retort), invert point is 30 ℃, rotating speed is 150r/min, after transforming 6h, with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), record in the fermented liquid 1 after treatment, 6-hexose diphosphate na concn is 5.11g/L.
Embodiment 6
Select plum mountain barms for use, 28-30 ℃, (substratum: pol is the wort of 12 Berlin to 150r/min shaking table culturing cell, pH6.5, the bottled 100mL of 500mL triangle), the 24h collecting cell obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10000r/min.
Get the centrifugal yeast cell 1g that obtains, adding mass percent is 10.0% glucose, 3.10% SODIUM PHOSPHATE, MONOBASIC, the 30mL reaction solution of pH6.5, invert point are 30 ℃, and rotating speed is 150r/min, after transforming 6h, with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), record in the fermented liquid 1 after treatment, 6-hexose diphosphate na concn is 3.57g/L.
Embodiment 7
Select plum mountain barms for use, 28-30 ℃, (substratum: pol is the wort of 12 Berlin to 150r/min shaking table culturing cell, pH6.5, the bottled 100mL of 500mL triangle), the 24h collecting cell obtained yeast cell in centrifugal 30 minutes under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 10000r/min.
Get the centrifugal yeast cell 1g that obtains, adding mass percent is 10.0% glucose, 3.10% SODIUM PHOSPHATE, MONOBASIC is added tween 80 0.05%, the 30mL reaction solution of pH6.5, invert point is 30 ℃, rotating speed is 150r/min, behind the conversion 6h, with 1mol/L Tricholroacetic Acid aqueous solution termination reaction (Tricholroacetic Acid amount of aqueous solution used 5ml), record in the fermented liquid 1 after treatment, 6-hexose diphosphate na concn is 5.57g/L.And the FDP sodium resultant quantity that does not add tween 80 in the same transformation system is 3.60g/L, 1, and the system that do not add tween 80 of 6-hexose diphosphate na concn under the similarity condition.
Claims (7)
1, a kind of yeast cell biosynthesizing 1, the method for 6-hexose diphosphate sodium comprises the steps:
In the reaction solution of 100mL sterilization, add 1～5g through the pre-thalline that obtains of cultivating, at 25～30 ℃, 100～200r/min conversion reaction, 4～10h, add the Tricholroacetic Acid termination reaction after, centrifugation obtains containing 1, the supernatant liquor of 6-hexose diphosphate sodium;
Described thalline is that yeast saccharomyces cerevisiae was cultivated 12～36 hours through pre-, again the cell that obtains through frozen centrifugation;
The pH5.0 of described reaction solution～9.0, the mass percent of reaction solution consists of:
2～12% glucose;
2～15% SODIUM PHOSPHATE, MONOBASIC;
Surplus is a water.
2, the method for claim 1 is characterized in that: in the reaction solution of 100mL sterilization, add the 3g thalline, at 30 ℃, 150r/min conversion reaction 6h, add the Tricholroacetic Acid termination reaction after, at 4 ℃, 10, the centrifugal 30min of 000r/min obtains containing 1, the supernatant liquor of 6-hexose diphosphate sodium.
3, the method for claim 1 is characterized in that: described reaction solution pH5.5～8.9, and the mass percent of reaction solution is formed preferred:
9%～11% glucose;
3%～7% SODIUM PHOSPHATE, MONOBASIC;
Surplus is a water.
4, the method for claim 1 is characterized in that: the mass percent of described reaction solution consists of:
9%～11% glucose;
3%～7% SODIUM PHOSPHATE, MONOBASIC;
0.05%～0.10% tween 80;
Surplus is a water.
5, the method for claim 1 is characterized in that: the fermention medium that described pre-cultivation is adopted is that pol is 12 Berlin, and pH is 6.5 wort.
6, method as claimed in claim 5 is characterized in that: described pre-cultivation is cultivated in advance at 25～30 ℃, 150r/min shaking table.
7, method as claimed in claim 6 is characterized in that: the condition of described frozen centrifugation is 4 ℃, 10000r/min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105733946A (en) * | 2015-12-30 | 2016-07-06 | 江南大学 | Production method of fructose 1,6-diphosphate (FDP) through low temperature spray drying and active dry yeast conversion |
| CN106222211A (en) * | 2016-07-21 | 2016-12-14 | 中国科学院天津工业生物技术研究所 | The preparation method of 1,6 fructose diphosphate |
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2008
- 2008-08-01 CN CNA2008100637940A patent/CN101328489A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105733946A (en) * | 2015-12-30 | 2016-07-06 | 江南大学 | Production method of fructose 1,6-diphosphate (FDP) through low temperature spray drying and active dry yeast conversion |
| CN105733946B (en) * | 2015-12-30 | 2019-08-06 | 江南大学 | A kind of method that low temperature spray drying prepares active dry yeast conversion 1,6 diphosphofructoses of production |
| CN106222211A (en) * | 2016-07-21 | 2016-12-14 | 中国科学院天津工业生物技术研究所 | The preparation method of 1,6 fructose diphosphate |
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