CN105274158A - Method for extracting polyglutamic acid from fermentation liquor through aqueous two-phase technology - Google Patents
Method for extracting polyglutamic acid from fermentation liquor through aqueous two-phase technology Download PDFInfo
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- CN105274158A CN105274158A CN201510817118.8A CN201510817118A CN105274158A CN 105274158 A CN105274158 A CN 105274158A CN 201510817118 A CN201510817118 A CN 201510817118A CN 105274158 A CN105274158 A CN 105274158A
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- fermented liquid
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Abstract
The invention discloses a method for extracting polyglutamic acid from fermentation liquor through the aqueous two-phase technology. The method comprises the steps that a mixed solution is firstly prepared, and an aqueous two-phase extraction system is built, so that the mass fraction of PEG1000 ranges from 10% to 30%, the mass fraction of (NH4)2SO4 ranges from 5% to 25%, the mass fraction of the fermentation liquor is 10%, the pH value of the fermentation liquor ranges from 3.75 to 8.46, and then environment temperature is controlled to range from 20 DEG C to 40 DEG C; after the mixed liquid is subjected to natural phase separation, an upper phase is taken, and PEG1000 penetrates through an ultrafiltration membrane with the molecular weight cutoff of 30 KDa; finally, after the cut-off polyglutamic acid concentrated solution is subjected to low-temperature refrigeration at the temperature of -40 DEG C, vacuum freeze drying is carried out for 24 h at the temperature of -50 DEG C, and a pure product is obtained. The problem that polyglutamic acid fermentation liquor is thick, and consequently pretreatment cost is too high is solved, the subsequent aqueous two-phase system reaction is moderate, and damage of a traditional precipitation method to polyglutamic acid molecular weight is avoided.
Description
Technical field
The present invention relates to a kind of method adopting Two-phase system to extract polyglutamic acid from fermented liquid, belong to bioseparation field of engineering technology.
Background technology
γ-PGA has excellent water-soluble, superpower adsorptivity and biodegradability, degraded product is non-harmful L-glutamic acid, it is a kind of excellent environment-friendly type macromolecule material, can be used as water-holding agent, heavy metal flocculant, sustained release dosage and pharmaceutical carrier etc., very large commercial value and social value is all had in industries such as makeup, environmental protection, food, medicine, agriculturals, but polyglutamic acid fermented liquid thickness, pre-treatment cost is excessive, and traditional precipitator method can damage polyglutamic acid molecular weight.
Summary of the invention
An object of the present invention is to solve above-mentioned polyglutamic acid fermented liquid thickness, pre-treatment cost is excessive, it is simple that the tradition precipitator method provide a kind of technique to problems such as polyglutamic acid molecular weight damage, cost is low, reaction temperature and double-aqueous phase system, avoid the destruction of polyglutamic acid molecular weight simultaneously.
Two of object of the present invention is to provide a kind of method adopting Two-phase system to extract polyglutamic acid from fermented liquid.
Technical scheme of the present invention is specifically described as follows.
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, namely first prepare mixing solutions, set up two phase aqueous extraction system, make PEG1000 massfraction be 10%-30%, (NH
4)
2sO
4massfraction is 5%-25%, fermented liquid massfraction is 10%, fermented liquid pH value is 3.75-8.46, then the temperature that controls environment is 20-40 DEG C, and solution nature to be mixed phase-splitting, gets phase, molecular weight cut-off is adopted to be that the ultra-filtration membrane of 30KDa is through PEG1000, finally by the polyglutamic acid concentrated solution that retains after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtain sterling.
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, be prepared from as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 10%-30%, (NH
4)
2sO
4massfraction is 5%-25%, fermented liquid massfraction is 10%, and fermented liquid pH value is 3.75-8.46;
(2) natural phase-splitting
The temperature that controls environment is 20-40 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
Beneficial effect of the present invention is as follows:
A kind of method adopting Two-phase system to extract polyglutamic acid from fermented liquid of the present invention, technique is simple, and cost is low, solves polyglutamic acid fermented liquid thickness, avoids the destruction of traditional precipitator method to polyglutamic acid molecular weight.
Embodiment
Below by specific embodiment, the present invention is set forth further, but do not limit the present invention.
A kind of Two-phase system that adopts of the present invention extracts polyglutamic acid fermented liquid used from fermented liquid, is made by oneself by laboratory.
A kind of Two-phase system that adopts of the present invention extracts polyglutamic acid small-sized tangential flow ultra-filtration unit used from fermented liquid, is produced by Millipore company of the U.S..
A kind of Two-phase system that adopts of the present invention extracts polyglutamic acid vacuum freeze drier used from fermented liquid, is produced by Beijing Bo Yikang laboratory apparatus company limited.
Embodiment 1
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, be prepared from as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 10%, (NH
4)
2sO
4massfraction is 5%, fermented liquid massfraction is 10%, and fermented liquid pH value is 3.75;
(2) natural phase-splitting
The temperature that controls environment is 20 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
The polyglutamic acid double-aqueous phase system of above-mentioned gained, partition ratio is 0.93, and extraction yield is 51.25%.
Embodiment 2
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, be prepared from as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 30%, (NH
4)
2sO
4massfraction is 25%, fermented liquid massfraction is 10%, and fermented liquid pH value is 8.46;
(2) natural phase-splitting
The temperature that controls environment is 40 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
The polyglutamic acid double-aqueous phase system of above-mentioned gained, partition ratio is 0.81, and extraction yield is 45.06%.
Embodiment 3
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, be prepared from as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 20%, (NH
4)
2sO
4massfraction is 15%, fermented liquid massfraction is 10%, and fermented liquid pH value is 5.31;
(2) natural phase-splitting
The temperature that controls environment is 30 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
The polyglutamic acid double-aqueous phase system of above-mentioned gained, partition ratio is 1.21, and extraction yield is 67.59%.
Embodiment 4
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, be prepared from as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 15%, (NH
4)
2sO
4massfraction is 10%, fermented liquid massfraction is 10%, and fermented liquid pH value is 4.66;
(2) natural phase-splitting
The temperature that controls environment is 25 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
The polyglutamic acid double-aqueous phase system of above-mentioned gained, partition ratio is 2.31, and extraction yield is 78.02%.
Embodiment 5
Adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, be prepared from as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 25%, (NH
4)
2sO
4massfraction is 20%, fermented liquid massfraction is 10%, and fermented liquid pH value is 7.55;
(2) natural phase-splitting
The temperature that controls environment is 35 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
The polyglutamic acid double-aqueous phase system of above-mentioned gained, partition ratio is 1.52, and extraction yield is 73.25%.
In sum, a kind of method adopting Two-phase system to extract polyglutamic acid from fermented liquid of the present invention, technique is simple, and cost is low, and its partition ratio is 0.81-2.31, and extraction yield is 45.06%-78.02%.Show thus, Two-phase system extracts polyglutamic acid and solves polyglutamic acid fermented liquid thickness from fermented liquid, the problem that pre-treatment cost is excessive, and follow-up double-aqueous phase system reaction temperature and, avoid the destruction of traditional precipitator method to polyglutamic acid molecular weight.
Above embodiment is only for illustration of content of the present invention; in addition; the present invention also has other embodiments, as long as those skilled in the art are because of technology involved in the present invention enlightenment, and the technical scheme adopting equivalent replacement or equivalent deformation mode to be formed all drops in protection scope of the present invention.
Claims (7)
1. adopt Two-phase system from fermented liquid, extract a method for polyglutamic acid, it is characterized in that, concrete steps are as follows:
(1) mixing solutions preparation
Preparation mixing solutions, sets up two phase aqueous extraction system, makes PEG1000 massfraction be 10%-30%, (NH
4)
2sO
4massfraction is 5%-25%, fermented liquid massfraction is 10%, and fermented liquid pH value is 3.75-8.46;
(2) natural phase-splitting
The temperature that controls environment is 20-40 DEG C, treats mixing solutions nature phase-splitting prepared by step (1);
(3) get the upper phase of step (2) mixed solution, adopt molecular weight cut-off be the ultra-filtration membrane of 30KDa through PEG1000, the polyglutamic acid concentrated solution retained, after-40 DEG C of cryogenic freezings, in-50 DEG C of vacuum lyophilization 24h, obtains sterling.
2. the method adopting Two-phase system to extract polyglutamic acid from fermented liquid as claimed in claim 1, it is characterized in that, in step (1), the proportioning of mixing solutions is: PEG1000 massfraction is 10%, (NH
4)
2sO
4massfraction is 5%, and fermented liquid massfraction is 10%, and fermented liquid pH value is 3.75; Natural phase-splitting described in step (2), the temperature that controls environment is 20 DEG C.
3. the method adopting Two-phase system to extract polyglutamic acid from fermented liquid as claimed in claim 1, it is characterized in that, in step (1), the proportioning of mixing solutions is: PEG1000 massfraction is 30%, (NH
4)
2sO
4massfraction is 25%, and fermented liquid massfraction is 10%, and fermented liquid pH value is 8.46; Natural phase-splitting described in step (2), the temperature that controls environment is 40 DEG C.
4. the method adopting Two-phase system to extract polyglutamic acid from fermented liquid as claimed in claim 1, it is characterized in that, in step (1), the proportioning of mixing solutions is: PEG1000 massfraction is 20%, (NH
4)
2sO
4massfraction is 15%, and fermented liquid massfraction is 10%, and fermented liquid pH value is 5.31; Natural phase-splitting described in step (2), the temperature that controls environment is 30 DEG C.
5. the method adopting Two-phase system to extract polyglutamic acid from fermented liquid as claimed in claim 1, is characterized in that, in step (1), the proportioning of mixing solutions is: PEG1000 massfraction is 15%, (NH
4)
2sO
4massfraction is 10%, and fermented liquid massfraction is 10%, and fermented liquid pH value is 4.66; Natural phase-splitting described in step (2), the temperature that controls environment is 25 DEG C.
6. the method adopting Two-phase system to extract polyglutamic acid from fermented liquid as claimed in claim 1, is characterized in that, in step (1), the proportioning of mixing solutions is: PEG1000 massfraction is 25%, (NH
4)
2sO
4massfraction is 20%, and fermented liquid massfraction is 10%, and fermented liquid pH value is 7.55; Natural phase-splitting described in step (2), the temperature that controls environment is 35 DEG C.
7. the employing Two-phase system as described in as arbitrary in claim 1-6 extracts the method for polyglutamic acid from fermented liquid, it is characterized in that, when extracting polyglutamic acid, its partition ratio is 0.81-2.31, and extraction yield is 45.06%-78.02%.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101503511A (en) * | 2009-03-06 | 2009-08-12 | 天津商业大学 | Method for extracting polyglutamic acid from original fermentation liquor by double aqueous phase system |
| CN102584933A (en) * | 2012-02-13 | 2012-07-18 | 汪志友 | Method for improving separation efficiency, purity and biological specific activity of blood coagulation factor VIII and analog thereof by using affine aqueous two-phase system |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101503511A (en) * | 2009-03-06 | 2009-08-12 | 天津商业大学 | Method for extracting polyglutamic acid from original fermentation liquor by double aqueous phase system |
| CN102584933A (en) * | 2012-02-13 | 2012-07-18 | 汪志友 | Method for improving separation efficiency, purity and biological specific activity of blood coagulation factor VIII and analog thereof by using affine aqueous two-phase system |
Non-Patent Citations (1)
| Title |
|---|
| 王雷: "采用双水相系统从发酵液中提取聚谷氨酸", 《食品研究与开发》 * |
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