CN104263787B - Method for removing fluoride and ash in euphausia superba polypeptide - Google Patents
Method for removing fluoride and ash in euphausia superba polypeptide Download PDFInfo
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- CN104263787B CN104263787B CN201410460399.1A CN201410460399A CN104263787B CN 104263787 B CN104263787 B CN 104263787B CN 201410460399 A CN201410460399 A CN 201410460399A CN 104263787 B CN104263787 B CN 104263787B
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Abstract
The invention aims at providing a method for removing fluoride and ash in euphausia superba polypeptide. The method comprises the following operating steps: hydrolyzing euphausia superb powder with protease to obtain an euphausia superba polypeptide aqueous solution; adjusting the content of solid substances in the polypeptide aqueous solution by virtue of dilution or concentration to obtain an euphausia superba polypeptide feedstock solution; continuously introducing the euphausia superba polypeptide feedstock solution into zones of a simulated moving bed so that the entire processes of adsorption, elution, regeneration and the balance are completed in an ion exchange column within a process cycle by virtue of the periodic switching of valves; and spray-drying the euphausia superba polypeptide solution from which fluorine and salt are removed to obtain high-quality euphausia superba polypeptide powder, wherein the simulated moving bed device adopts the ion exchange column as an operating unit and at least comprises five zones, namely, a feed zone, an elution zone, a cation regeneration zone, an anion regeneration zone and a balance zone. The method has the advantages of small device structure, high yield, low solvent consumption, high separation capacity, less pollution and the like and by virtue of the method, the continuous large-scale industrial production and automatic control are achieved.
Description
Technical field
The invention belongs to polypeptide extraction purification technical field, be specifically related to a kind of method removing fluorochemical and ash content in krill polypeptide.
Background technology
Krill is a class marine invertebrate, is distributed in antarctic waters.According to statistics, the standing stock of krill are 6.5-10 hundred million tons, are the huge protein resource treasure-houses of the mankind.In krill dry weight, protein content is about 65-75%, and its amino acid composition is reasonable, essential amino acids content is high, is the important source material of carrying out protein higher value application.
Krill polypeptide adopts krill to be raw material, prepares the soluble small molecular polypeptide of gained through protease hydrolysis.The ash oontent being processed the krill meal obtained by whole shrimp is generally 10 ~ 17%, and Oil repellent is generally 1800-2500ppm.Take krill meal as raw material again, prepared in the process of krill polypeptide by protease hydrolysis, ash contents a large amount of in shrimp med and fluorochemical can remain in polypeptide products.Meanwhile, need to add the acid-basicity that acid or alkali carrys out maintenance system in enzymolysis process and stablize, thus cause the ash oontent in krill polypeptide products to raise further.And the high fluoride of krill polypeptide and high ash oontent, have a strong impact on the development and application prospect of polypeptide as functional protein food.
At present, the polypeptide desalinating process reported mainly comprises membrane processing method (ultrafiltration, micro-filtration, nanofiltration), electroosmose process, single-column ion exchange method and Amberlyst process, and first three plants technique is grow up on the basis of water desalination technology.Polypeptide is organism, and in the solution with certain electric charge, film process desalination process film very easily occurs by the phenomenon of Organic pollutants, makes the lost of life of film, and cost increases.Electroosmose process desalination can cause larger polypeptide loss, thus makes the rate of recovery on the low side, and the cost of electroosmose process is higher.The wash-out link of Amberlyst process needs with an organic solvent, not easily realizes industrialization.Single-column ion exchange method has that the rate of recovery is high, ratio of desalinization advantages of higher, but long processing period and more bronsted lowry acids and bases bronsted lowry can be used.
Simulation moving-bed (SMB) technology is switched by the regular order of stream socket position, fixed bed device simulates stationary phase reverse circulation flowing relative to moving phase, make separation system obtain larger mass transfer force, high-efficiency and continuous carries out a kind of technology of lock out operation.SMB technology has the advantages such as device structure is little, productive rate is high, solvent consumption is few, separating power is strong, pollution is few.SMB technology and ion-exchange are used in conjunction the periodical operation overcoming ADSORPTION IN A FIXED BED isolation technique and exist, the shortcoming that solvent consumption is large, product concentration is low, achieve continuity large-scale industrial production and automatically control.Current SMB technology has been applied in the separating-purifying of biology, chiral drug, fine chemicals and food.
Foreign patent EP1106602B1 discloses the method that adopts the acid of SMB technology continuous purifying ammonia base.Domestic patent CN102050865B discloses a method by SMB technology chromatography purified polypeptide, is separated by immunoreactivity compound with other non-intrinsically safe organic compositions.Domestic patent CN103787868A discloses the method that adopts four district SMB technology separation succsinic acids and lactic mixt.Liu Jiangli (the industrialization technology parameter study of Lactoferrin from bovine colostrums separation and purification, Heilongjiang Bayi Agricultural Reclamation University, academic dissertation, 2011) adopts SMB technology continuous separation technology to carry out separation and purification to lactoferrin, and purity is 95.8%.Adopt the method process krill polypeptide of existing SMB separation and purification proteins and peptides, the object of fluorochemical and ash content in the water-soluble krill polypeptide of effective elimination can not be reached, meanwhile, there is not yet both at home and abroad at present SMB technology is applied to the relevant report removing fluorochemical and ash content.
Summary of the invention
The object of this invention is to provide a kind of method removing fluorochemical and ash content in krill polypeptide, thus make up the deficiencies in the prior art.
Remove the method for fluorochemical and ash content in krill polypeptide in the present invention, include following step:
1) in euphausia superba powder, add distilled water, wherein the mass ratio of krill meal and water is 1:6 ~ 15, regulates mixed solution pH value, then in mixed solution, adds proteolytic enzyme be hydrolyzed, and hydrolysis temperature is 40 ~ 60 DEG C, and hydrolysis time is 2 ~ 12h; Hydrolysis terminates post-heating enzymolysis reaction, centrifugal, obtains supernatant liquor, is krill polypeptid solution;
2) prepare simulation moving-bed device, simulation moving-bed device includes intake zone I, elution zone II, positively charged ion breeding blanket III, anion regeneration district IV, equilibrium area V; Wherein, elution zone is positioned in the middle of positively charged ion breeding blanket and intake zone; Anion regeneration district is positioned in the middle of equilibrium area and positively charged ion breeding blanket; Intake zone is positioned in the middle of elution zone and equilibrium area; Wherein intake zone is alternately composed in series with the anion-exchange column being no less than 3 by the cationic exchange coloum being no less than 3, elution zone is made up of with the anion-exchange column being no less than 1 cationic exchange coloum being no less than 1, positively charged ion breeding blanket is made up of the cationic exchange coloum being no less than 1, anion regeneration district is made up of the anion-exchange column being no less than 1, and equilibrium area is made up of with the anion-exchange column being no less than 1 cationic exchange coloum being no less than 1;
3) krill polypeptid solution is introduced continuously the import in simulation moving-bed I district, remove the product after fluorochemical and ash content and flowed out by the outlet in I district; The elutriant in I district is flowed into by the import of II district, and the elutriant containing krill polypeptide that II district flows out imports I district again to carry out removing fluorochemical and ash treatment again; Wherein the ion column in I district and II district is undertaken regenerating and Balance Treatment by III district, IV district and V district successively; Ion column carries out regeneration activating in III district and IV district regenerated liquid, and in V district, balance liquid carries out balance purge; The balance liquid that V district uses is deionized water; The ion regenerated liquid in III district is 2% ~ 4%HCl solution, and IV district ion regenerated liquid is 2% ~ 4%NaOH solution;
4) the krill polypeptide solution removing fluorochemical and ash content is spray-dried, obtains krill polypeptide powder;
Wherein, step 1) in lipid content in euphausia superba powder be 1-20%; Proteolytic enzyme is one or more in food grade papoid, bromeline, trypsinase, neutral protease or Sumizyme MP;
Step 2) in the number of intake zone and elution zone be two or more;
Step 2) resin is strong acid type or weakly acidic cation-exchange resin in cationic exchange column, in anion-exchange column, resin is strong basicity or weak base anion-exchange resin;
Step 3) in krill propolypeptide feed liquid contact with the solid phase of ion exchange column in a counter-current configuration, charging flow velocity is 1 ~ 8BV/h, and elution flow rate is 1 ~ 8BV/h, and regeneration velocity is 1 ~ 8BV/h, and equilibrium velocity is 1 ~ 8BV/h;
Step 4) in spray-dired condition as follows: inlet temperature is 120 ~ 160 DEG C, and air outlet temperature is 80 ~ 120 DEG C;
Be 350 ~ 450mg/kg without Oil repellent in the krill polypeptide powder removing fluorochemical and ash content in the present invention, ash oontent is 7 ~ 15%; Removing Oil repellent in the krill polypeptide after fluorochemical and ash content through SMB technology is 2 ~ 10mg/kg, ash oontent≤4%, protein content >=90%.The present invention, by the regular switching of valve, makes exchange column in a technological cycle, complete all processes of absorption, wash-out, regeneration, balance.The method has the advantages such as device structure is little, productive rate is high, solvent consumption is few, separating power is strong, pollution is few, achieves continuity large-scale industrial production and automatically controls.
Accompanying drawing explanation
Fig. 1: be a kind of simulation moving-bed setting drawing removing fluorochemical and ash content in krill polypeptide of the present invention; Wherein, the exchange column that shade is filled is cationic exchange coloum; Blank exchange column of filling is anion-exchange column;
Fig. 2: the setting drawing removing fluorochemical and ash content in krill polypeptide for another kind of the present invention is simulation moving-bed, wherein, the exchange column that shade is filled is cationic exchange coloum; Blank exchange column of filling is anion-exchange column.
Embodiment
The invention provides the method for fluorochemical and ash content in the simultaneous removing krill polypeptide of a kind of easy and simple to handleization, technique serialization, with low costization.The method is with short production cycle, desalination degree is high, product recovery rate and purity high, provide theoretical foundation for being applied to large-scale industrial production.
Simulation moving-bed device of the present invention comprises intake zone I, elution zone II, positively charged ion breeding blanket III, anion regeneration district IV and equilibrium area V five regions.The ion exchange column (cationic exchange coloum and anion-exchange column) of two types is comprised in simulation moving-bed device.Wherein, elution zone is positioned in the middle of positively charged ion breeding blanket and intake zone; Anion regeneration district is positioned in the middle of equilibrium area and positively charged ion breeding blanket; Intake zone is positioned in the middle of elution zone and equilibrium area; Wherein intake zone is alternately composed in series with the anion-exchange column being no less than 3 by the cationic exchange coloum being no less than 3, elution zone is made up of with the anion-exchange column being no less than 1 cationic exchange coloum being no less than 1, positively charged ion breeding blanket is made up of the cationic exchange coloum being no less than 1, anion regeneration district is made up of the anion-exchange column being no less than 1, and equilibrium area is made up of with the anion-exchange column being no less than 1 cationic exchange coloum being no less than 1.Switch by the regular of different valve the inflow and outflow that control stock liquid, elutriant, regenerated liquid and balance liquid, reach the object removing fluorochemical and ash content in propolypeptide feed liquid.
When krill propolypeptide feed liquid flows into I district, organic polypeptide fraction can be made to separate with inorganic component (fluorochemical and ash content).In ion exchange column, the adsorptive power of polypeptide fraction is more weak, and the zwitterion in inorganic component exchanges respectively in anion-exchange column and cationic exchange coloum.Polypeptide fraction flows out I district prior to inorganic component.Collect the product that purity is higher, product lower for purity is imported again the process that I district carries out removing fluorochemical and ash content.Elutriant flows into II district, is washed out by the polypeptide fraction remained in pillar gap, and the elutriant containing polypeptide fraction imports I district.Two strands of different regenerated liquids flow into III district and IV district respectively, carry out manipulation of regeneration to ion exchange column, regenerated liquid after reverse osmosis for the preparation of stock liquid krill polypeptide.Balance liquid flows into V district, is washed out by regenerated liquid remaining in ion exchange column, and balance flow fluid imports regenerated liquid.
(illustrate for Fig. 1) when current period switches, all pillars move two respectively to the reverse direction that fluid flows, and 7, No. 8 pillars in I district enter II district; 5, No. 6 pillars in II district enter III district and the regeneration of IV district respectively; 4, No. 3 pillars in III district and IV district enter V district and balance; 1, No. 2 pillar in V district enters I district, becomes last two pillars in I district.
Method of the present invention, by increasing the repeat region of subregion or the exchange column number of subregion, can improve turnout.
Embodiment 1
1) euphausia superba powder 5kg is got, distilled water 60L is added in krill meal, 2M HCl is added in mixed solution, regulate the pH value to 7.5 of mixed solution, be warming up to 45 DEG C, in mixed solution, add neutral protease 75g, constant temperature stirs 10h in a kettle., be heated to 100 DEG C and stop enzyme reaction, centrifugal, obtain supernatant liquor; Make the solid content of supernatant liquor reach 10% by vacuum concentration, vacuum tightness is 0.08MPa, and temperature is 40 ~ 80 DEG C, obtains krill propolypeptide feed liquid;
2) the simulation moving-bed device of 12 exchange columns as shown in Figure 1, is selected; Elutriant is deionized water; Anion regeneration liquid 1 is 2% ~ 4%NaOH solution, and positively charged ion regenerated liquid 2 is 2% ~ 4%HCl solution; Balance liquid is deionized water;
3) the charging flow velocity of stock liquid is 4BV/h; The flow velocity of elutriant is 2.4BV/h; The flow velocity of regenerated liquid 1 and regenerated liquid 2 is 1.6BV/h; The flow velocity of balance liquid is 1.6BV/h; Switching time is 75min;
4) collect the high-purity product flowed out in I district, after spraying dry (inlet temperature 120 DEG C, air outlet temperature 80 DEG C), obtain low fluorine, low-ash high-quality krill polypeptide powder.
Be 350 ~ 450mg/kg without Oil repellent in the krill polypeptide powder removing fluorochemical and ash content in the present invention, ash oontent is 7 ~ 15%; Removing Oil repellent in the krill polypeptide after fluorochemical and ash content through SMB technology is 2 ~ 10mg/kg, ash oontent≤4%, protein content >=90%, protein recovery >=80%.
Embodiment 2
1) get euphausia superba powder 10kg, add 95% ethanol 100L, stir 6h in a kettle. in krill meal, centrifuging and taking precipitates, and vacuum-drying obtains degreasing krill meal;
2) extracting degreasing shrimp med 5kg, distilled water 50L is added in degreasing shrimp med, 2M HCl is added in mixed solution, regulate mixed solution pH value to 8.0, be warming up to 50 DEG C, in mixed solution, add Sumizyme MP 80g, constant temperature stirs 8h in a kettle., be heated to 100 DEG C and stop enzyme reaction, centrifugal, obtain supernatant liquor; Make the solid content of supernatant liquor reach 20% by vacuum concentration, vacuum tightness is 0.08MPa, and temperature is 40 ~ 80 DEG C, obtains krill propolypeptide feed liquid;
3) the simulation moving-bed device of 20 exchange columns as shown in Figure 2, is selected; Elutriant is deionized water; Regenerated liquid 1 is 2% ~ 4%NaOH solution, and regenerated liquid 2 is 2% ~ 4%HCl solution; Balance liquid is deionized water;
4) the charging flow velocity of stock liquid is 3BV/h, and 13 ~ No. 16 pillar flow velocitys in I district are 3BV/h, and 17 ~ No. 20 pillar flow velocitys are 6BV/h; The flow velocity of elutriant is 5BV/h; The flow velocity of regenerated liquid 1 and regenerated liquid 2 is 3BV/h; The flow velocity of balance liquid is 3BV/h; Switching time is 60min;
5) collect the high-purity product flowed out in I district, after spraying dry (inlet temperature 130 DEG C, air outlet temperature 85 DEG C), obtain low fluorine, low-ash high-quality krill polypeptide powder.
Be 350 ~ 450mg/kg without Oil repellent in the krill polypeptide powder removing fluorochemical and ash content in the present invention, ash oontent is 7 ~ 15%; Removing Oil repellent in the krill polypeptide after fluorochemical and ash content through SMB technology is 2 ~ 10mg/kg, ash oontent≤4%, protein content >=90%, protein recovery >=80%.Result shows, after I district and II district increase exchange column, switching time shortens 15min, and the treatment capacity in the unit time adds one times, product purity and yield unaffected.
Claims (5)
1. remove a method for fluorochemical and ash content in krill polypeptide, it is characterized in that, described method includes following step:
1) in euphausia superba powder, add distilled water, wherein the mass ratio of krill meal and water is 1:6 ~ 15, regulates mixed solution pH value, then in mixed solution, adds proteolytic enzyme be hydrolyzed, and hydrolysis temperature is 40 ~ 60 DEG C, and hydrolysis time is 2 ~ 12h; Hydrolysis terminates post-heating enzymolysis reaction, centrifugal, obtains supernatant liquor, is krill polypeptid solution;
2) prepare simulation moving-bed device, simulation moving-bed device includes intake zone I, elution zone II, positively charged ion breeding blanket III, anion regeneration district IV, equilibrium area V; Wherein, elution zone is positioned in the middle of positively charged ion breeding blanket and intake zone; Anion regeneration district is positioned in the middle of equilibrium area and positively charged ion breeding blanket; Intake zone is positioned in the middle of elution zone and equilibrium area; Wherein intake zone is alternately composed in series with the anion-exchange column being no less than 3 by the cationic exchange coloum being no less than 3, elution zone is made up of with the anion-exchange column being no less than 1 cationic exchange coloum being no less than 1, positively charged ion breeding blanket is made up of the cationic exchange coloum being no less than 1, anion regeneration district is made up of the anion-exchange column being no less than 1, and equilibrium area is made up of with the anion-exchange column being no less than 1 cationic exchange coloum being no less than 1;
Described intake zone and the number of elution zone are two or more;
In described cationic exchange coloum, resin is strong acid type or weakly acidic cation-exchange resin, and the resin of anion-exchange column is strong basicity or weak base anion-exchange resin;
3) krill polypeptid solution is introduced continuously the import in simulation moving-bed I district, remove the product after fluorochemical and ash content and flowed out by the outlet in I district; The elutriant in I district is flowed into by the import of II district, and the elutriant containing krill polypeptide that II district flows out imports I district again to carry out removing fluorochemical and ash treatment again; Wherein the ion column in I district and II district is undertaken regenerating and Balance Treatment by III district, IV district and V district successively; Ion column carries out regeneration activating in III district and IV district regenerated liquid, and in V district, balance liquid carries out balance purge;
Wherein krill polypeptid solution contacts with the solid phase of ion exchange column in a counter-current configuration, and charging flow velocity is 1 ~ 8BV/h, and elution flow rate is 1 ~ 8BV/h, and regeneration velocity is 1 ~ 8BV/h, and equilibrium velocity is 1 ~ 8BV/h;
4) the krill polypeptide solution removing fluorochemical and ash content is spray-dried, obtains krill polypeptide powder.
2. the method for claim 1, is characterized in that, described step 1) in lipid content in euphausia superba powder be 1-20%; Proteolytic enzyme is one or more in food grade papoid, bromeline, trypsinase, neutral protease or Sumizyme MP.
3. the method for claim 1, is characterized in that, described step 3) the ion regenerated liquid in Zhongⅲ district is 2% ~ 4%HCl.
4. the method for claim 1, is characterized in that, described step 3) the ion regenerated liquid in Zhongⅳ district is 2% ~ 4%NaOH.
5. the method for claim 1, is characterized in that, described step 4) in spray-dired condition as follows: inlet temperature is 120 ~ 160 DEG C, and air outlet temperature is 80 ~ 120 DEG C.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4091620A1 (en) * | 2018-01-30 | 2022-11-23 | Aker Biomarine Antarctic As | Krill protein hydrolysate with low fluoride and trimethylamin content |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105265998A (en) * | 2015-09-15 | 2016-01-27 | 广东海洋大学 | Preparation method of antarctic krill low-fluoride enzymatic hydrolysate |
| CN109225355B (en) * | 2018-11-13 | 2023-11-14 | 赛普特环保技术(厦门)有限公司 | Continuous ion exchange process for removing inorganic salt and system adopted by same |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1777435A (en) * | 2002-09-13 | 2006-05-24 | 拜奥根Idec公司 | Method of purifying polypeptides by simulated moving bed chromatography |
| CN102485016A (en) * | 2010-12-06 | 2012-06-06 | 中国水产科学研究院东海水产研究所 | Preparation method of euphausia superba protein peptide mixture |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1777435A (en) * | 2002-09-13 | 2006-05-24 | 拜奥根Idec公司 | Method of purifying polypeptides by simulated moving bed chromatography |
| CN102485016A (en) * | 2010-12-06 | 2012-06-06 | 中国水产科学研究院东海水产研究所 | Preparation method of euphausia superba protein peptide mixture |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4091620A1 (en) * | 2018-01-30 | 2022-11-23 | Aker Biomarine Antarctic As | Krill protein hydrolysate with low fluoride and trimethylamin content |
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