CN111892498A - Method for extracting L-malic acid - Google Patents
Method for extracting L-malic acid Download PDFInfo
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- CN111892498A CN111892498A CN202010859506.3A CN202010859506A CN111892498A CN 111892498 A CN111892498 A CN 111892498A CN 202010859506 A CN202010859506 A CN 202010859506A CN 111892498 A CN111892498 A CN 111892498A
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- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 title claims abstract description 306
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 title claims abstract description 157
- 235000011090 malic acid Nutrition 0.000 title claims abstract description 156
- 229940116298 l- malic acid Drugs 0.000 title claims abstract description 150
- 238000000034 method Methods 0.000 title claims abstract description 35
- 239000011347 resin Substances 0.000 claims abstract description 42
- 229920005989 resin Polymers 0.000 claims abstract description 42
- 238000000926 separation method Methods 0.000 claims abstract description 42
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims abstract description 33
- 238000000855 fermentation Methods 0.000 claims abstract description 26
- 230000004151 fermentation Effects 0.000 claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 26
- 239000000047 product Substances 0.000 claims abstract description 23
- 230000020477 pH reduction Effects 0.000 claims abstract description 22
- 239000012535 impurity Substances 0.000 claims abstract description 21
- 238000001035 drying Methods 0.000 claims abstract description 20
- 238000001914 filtration Methods 0.000 claims abstract description 14
- 235000006408 oxalic acid Nutrition 0.000 claims abstract description 11
- 239000000706 filtrate Substances 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 238000001816 cooling Methods 0.000 claims description 15
- 239000013078 crystal Substances 0.000 claims description 15
- 238000002425 crystallisation Methods 0.000 claims description 15
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- 229920001897 terpolymer Polymers 0.000 claims description 8
- 230000005484 gravity Effects 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
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- 239000012141 concentrate Substances 0.000 claims description 3
- 239000012467 final product Substances 0.000 claims 1
- 238000007873 sieving Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 14
- 230000008569 process Effects 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 6
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- 238000004519 manufacturing process Methods 0.000 abstract description 4
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- 239000002994 raw material Substances 0.000 description 8
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- 239000012065 filter cake Substances 0.000 description 6
- 229940099690 malic acid Drugs 0.000 description 6
- 239000001630 malic acid Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 159000000007 calcium salts Chemical class 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 4
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- 230000008020 evaporation Effects 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 150000001450 anions Chemical class 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 239000001362 calcium malate Substances 0.000 description 2
- 229940016114 calcium malate Drugs 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- 238000005342 ion exchange Methods 0.000 description 2
- 239000003456 ion exchange resin Substances 0.000 description 2
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- 239000012528 membrane Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
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- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- 208000037157 Azotemia Diseases 0.000 description 1
- 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 description 1
- 206010019663 Hepatic failure Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 108010093096 Immobilized Enzymes Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 239000013543 active substance Substances 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
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- 235000008452 baby food Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- OLOZVPHKXALCRI-UHFFFAOYSA-L calcium malate Chemical compound [Ca+2].[O-]C(=O)C(O)CC([O-])=O OLOZVPHKXALCRI-UHFFFAOYSA-L 0.000 description 1
- 235000011038 calcium malates Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000003763 carbonization Methods 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 230000009246 food effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- HHLFWLYXYJOTON-UHFFFAOYSA-N glyoxylic acid Chemical compound OC(=O)C=O HHLFWLYXYJOTON-UHFFFAOYSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
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- 208000033065 inborn errors of immunity Diseases 0.000 description 1
- 239000000077 insect repellent Substances 0.000 description 1
- 125000003010 ionic group Chemical group 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 208000007903 liver failure Diseases 0.000 description 1
- 231100000835 liver failure Toxicity 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000003264 margarine Substances 0.000 description 1
- 235000013310 margarine Nutrition 0.000 description 1
- -1 medical care Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000008558 metabolic pathway by substance Effects 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
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- 231100000331 toxic Toxicity 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/48—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/43—Separation; Purification; Stabilisation; Use of additives by change of the physical state, e.g. crystallisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/47—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Crystallography & Structural Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention relates to an extraction method of L-malic acid, belonging to the technical field of biological fermentation downstream extraction of organic acid. The method for extracting the L-malic acid comprises the following steps: 1) adding oxalic acid into the L-malic acid fermentation liquor for acidification, and filtering to obtain a filtrate, namely an L-malic acid acidification liquor; 2) separating and removing impurities from the L-malic acid acidized solution through TY019 resin to obtain an L-malic acid separated solution; 3) concentrating and crystallizing the L-malic acid separation liquid obtained in the step 2) to obtain an L-malic acid wet product; 4) and (3) placing the wet L-malic acid product in a vacuum drying oven, and drying in a vacuum gradient manner to obtain a finished L-malic acid product. The TY019 resin is used for separating the L-malic acid acidized solution, the impurity removal effect is good, and the purity of the separated solution is high. Compared with the conventional L-malic acid extraction method, the method has the advantages of high yield and purity of the obtained L-malic acid product, greatly simplified operation process and greatly reduced production cost.
Description
Technical Field
The invention belongs to the technical field of organic acid biological fermentation downstream extraction, and particularly relates to an extraction method of L-malic acid.
Background
L-malic acid is widely used as a food additive and a functional food having excellent properties in the fields of foods, health products, medical care, cosmetics, and the like. L-malic acid is generally used as a sour agent, a pH adjuster, and an antioxidant synergist in various foods such as alcoholic beverages, candies, jams and jellies, margarine, soy sauce, vinegar, pickles, and the like. The L-malic acid has the functions of resisting fatigue and protecting liver, kidney and heart, can also be used in health products and pharmaceutical industry, such as medicinal preparation, tablet, syrup, amino acid solution and insect repellent, can be used for treating various diseases such as liver disease, anemia, hypoimmunity, uremia, hypertension, liver failure, etc., and can alleviate the toxic effect of anticancer drugs on normal cells. In the daily chemical industry and the cosmetics industry, the L-malic acid can be used for toothpaste, shampoo, high-grade special detergent, skin care products and the like. In industrial industry, L-malic acid can be used as resin curing agent, industrial cleaning agent and synthetic material plasticizer.
L-malic acid is an important organic acid generated in the metabolic process of organisms, is an important intermediate product in the metabolic process of tricarboxylic acid cycle and glyoxylate cycle branches of the tricarboxylic acid cycle, is an intermediate product of CO2 fixed reaction, is an active substance existing in organism cells, is easy to absorb, can rapidly pass through cell membranes, enters mitochondria to directly participate in energy metabolism, and can be directly used as an energy substance to be utilized by organisms. Research shows that ATP generation can be obviously improved by supplementing L-malic acid. This is also the reason why L-malic acid is preferred in developed western countries, for example, it is well established in the United states that DL-malic acid cannot be used in infant food, beverages and medicines, and L-malic acid must be used. In view of the effect of food on life maintenance, it is essentially involved in a metabolic and energy conversion process, and proteins, lipids, carbohydrates, etc. are finally passed through the tricarboxylic acid cycle, which is the last step of the conversion process and is the most important step, and it is related to whether the physiological functions of human body are normal. The health care effect of the L-malic acid is to prevent abnormal tricarboxylic acid cycle caused by the deficiency of the L-malic acid in human body, which leads to metabolic disorder. The L-malic acid is in a special position in a substance metabolism path, can directly participate in human metabolism, is directly absorbed by a human body, can provide energy to the human body in a short time and eliminate fatigue, has the functions of resisting fatigue and quickly restoring physical strength, and can be applied to functional foods.
The production method of L-malic acid includes direct extraction method, chemical synthesis method, immobilized enzyme or cell conversion method, one-step fermentation method and two-step fermentation method. In recent years, with the gradual importance of environmental protection, greenness and pure nature, the fermentation method is increasingly popular. In the prior art, an invention patent with application publication number CN103642853A discloses a new process for extracting L-malic acid, which comprises the following steps: (1) acidifying malic acid fermentation liquor with phosphoric acid, adjusting pH to 1.5, standing for 6h, and filtering with microfiltration membrane and ultrafiltration membrane to remove impurities to obtain L-malic acid mother liquor; neutralizing the L-malic acid mother liquor obtained in the step (1) with calcium carbonate until the pH value is 4.1-4.4, and filtering to obtain calcium malate; adding 1-3 times of water to adjust L-calcium malate into slurry, adding phosphoric acid, adjusting pH to about 1.5, and performing acidolysis to obtain acidolysis solution; filtering, and decolorizing the filtrate to obtain L-malic acid eluate; (3) after the L-malic acid eluent obtained in the step (2) passes through ion exchange resin, concentrating at low temperature by a cooling evaporator until white crystals appear, quickly transferring the feed liquid into a crystallizer, and slowly cooling for crystallization; the ion exchange resin is strong acid type cation exchange resin or strong base anion exchange resin, and the particle size is 0.3-1.2 mm; the evaporation temperature of the evaporator is controlled at 50-60 ℃, and the evaporation capacity is 0.5-1.5 m/h; the temperature reduction speed is 0.2-1 ℃/min in the cooling crystallization process, the final crystallization temperature is 0-20 ℃, and the constant temperature crystallization is carried out for 0.5-1 h; (4) separating, washing and drying the product. The process can be abbreviated as follows: the method comprises the steps of L-malic acid fermentation liquid → primary acidification → neutralization → secondary acidification → ion exchange → evaporation concentration → crystallization → wet product separation → drying, and has the disadvantages of complex extraction method, low yield and purity of the obtained L-malic acid, high cost, large water consumption in the steps of secondary acidification and ion exchange, serious waste of water resources, and environmental pollution caused by the steps of neutralization and secondary acidification.
Disclosure of Invention
The invention aims to provide an extraction method of L-malic acid, which solves the problems of complex process, low extraction purity and low yield of the traditional extraction method. The extraction method has the advantages of good separation and purification effects, simple process flow, convenient operation and high yield and purity of the obtained L-malic acid.
In order to achieve the above purpose, the invention adopts the technical scheme that:
the method for extracting the L-malic acid comprises the following steps:
1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification, and filtering to obtain a filtrate, namely an L-malic acid acidification liquor;
2) removing impurities: passing the L-malic acid acidified liquid obtained in the step 1) through a separation resin column to obtain an L-malic acid separation liquid;
3) concentration and crystallization: concentrating and crystallizing the L-malic acid separation liquid obtained in the step 2) to obtain an L-malic acid wet product;
4) gradient drying: and (3) drying the wet L-malic acid product in a vacuum gradient manner to obtain a finished L-malic acid product.
Further, adding oxalic acid into the L-malic acid fermentation liquor in the step 1) to acidify until the pH value is 1.5-3.0.
Further, the filter cake obtained from the filtration was washed with water to pH 5 in step 1) to ensure no loss of L-malic acid.
Further, the separation resin column in the step 2) is a TY019 separation resin column, and the TY019 resin is a terpolymer synthesized by taking EA, BA and TYH-SFLP-Buty1 as raw materials.
Further, when the resin column is separated in the step 2), the separated liquid is collected in sections, and the water in the gaps of the resin column is collected and separated in the first section and recycled; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and collecting the high-purity L-malic acid solution in the fourth stage, namely the L-malic acid separation solution.
Further, the concentration in the step 2) is to concentrate the L-malic acid separation liquid to the specific gravity of 1.2-1.5.
Further, the concentration in the step 2) is to concentrate the L-malic acid separation solution at the temperature of 40-80 ℃ under reduced pressure.
Further, the crystallization in the step 3) is to stir at the temperature of 30-60 ℃ for crystallization, add seed crystals, grow the crystals for 1h, cool to 20 ℃ at the speed of 3 ℃/h, then cool to 0-10 ℃ at the speed of 5 ℃/h in a gradient manner, and perform vacuum filtration or centrifugal separation on solid and liquid.
Further, the gradient drying in the step 4) is to cool and pump the wet L-malic acid product at normal temperature until the water content is below 2%, then heat up at a speed of 8 ℃/h in a gradient manner until the temperature is 55 ℃, dry for 3-5h, and screen to obtain the finished product.
The invention has the beneficial effects that:
the extraction method of L-malic acid of the invention, separate and remove the impurity with TY019 resin specially developed for L-malic acid acidizing fluid, this resin is a terpolymer synthesized by EA, BA and TYH-SFLP-Buty1 as raw materials, and introduce anion, cation monomer to carry on graft copolymerization while resin is synthesized, form the three-dimensional network structure, can combine with water molecule through solvation of ionic group in copolymer, form the dense hydrated layer on the surface, have very good characteristic of adsorbing protein, the non-ionic monomer in the resin has very good salt absorption effect at the same time, so can remove protein and salt impurity in L-malic acid acidizing fluid very well, so TY019 resin has high impurity removal rate to fermented L-malic acid acidizing fluid, the separation effect is good, the L-malic acid seperated fluid obtained is high in purity, The yield is high.
Compared with the common sulfuric acid or phosphoric acid, the method for extracting the L-malic acid has better acidification effect by using the oxalic acid to acidify the fermentation liquor, has less impurities in the acidified liquor after acidification and filtration, and can avoid the carbonization phenomenon possibly caused by using the sulfuric acid.
Compared with the conventional L-malic acid extraction method, the L-malic acid extraction method has the advantages that the yield and the purity of the L-malic acid product obtained by the method are high, the operation process is greatly simplified, and the production cost is greatly reduced.
Detailed Description
The following will further describe the present invention in conjunction with examples.
The L-malic acid fermentation liquor is prepared by fermenting Aspergillus oryzae as a production strain and glucose and calcium carbonate as raw materials in a culture medium at 20-50 ℃ for 40-160h to obtain 20L of L-malic acid fermentation liquor.
Example 1
The method for extracting the L-malic acid comprises the following steps:
1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification until the pH value is 2.0, carrying out vacuum filtration to remove calcium salt, mycelia and fermentation residues, adding deionized water into a filter cake obtained by filtration, and washing until the pH value is 5.0 so as to ensure that no malic acid is lost, thereby obtaining L-malic acid acidification liquor.
2) Removing impurities: feeding the L-malic acid acidized solution into a TY019 separation resin column at the temperature of 35 ℃, wherein the flow rate of the column is 50mL/min, the feeding amount is 150g, collecting the separation solution in sections, collecting water in gaps of chromatographic resin in the first section, and recycling the water; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and the fourth stage collects high-purity L-malic acid solution, namely L-malic acid separating solution. The TY019 resin is a terpolymer synthesized by taking EA, BA and TYH-SFLP-Buty1 as raw materials.
3) Concentration and crystallization: concentrating the L-malic acid separation liquid at 60 deg.C under reduced pressure until the specific gravity is 1.35, stirring at 50 deg.C for 30min, adding seed crystal, growing crystal for 1h, cooling to 20 deg.C at 3 deg.C/h, then cooling to 10 deg.C at 5 deg.C/h, and centrifuging to separate solid and liquid to obtain wet L-malic acid.
4) Drying: placing the wet L-malic acid in a vacuum drying oven, firstly pumping at normal temperature for 1h by a pump until the water content is below 2%, then heating at a speed of 8 ℃/h in a gradient manner until the temperature is up to 55 ℃, and drying in vacuum for 3h to obtain the finished L-malic acid product with the purity of 99.8%.
Example 2
The method for extracting the L-malic acid comprises the following steps:
1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification until the pH value is 1.8, carrying out vacuum filtration to remove calcium salt, mycelia and fermentation residues, adding deionized water into a filter cake obtained by filtration, washing until the pH value is 5.0, so as to ensure that no malic acid is lost, and obtaining L-malic acid acidification liquor.
2) Removing impurities: at the temperature of 35 ℃, enabling the L-malic acid acidized solution to enter a TY019 separation resin column, enabling the flow rate of the column to be 80mL/min, enabling the feeding amount to be 100g, collecting the separation solution in a segmented mode, collecting water in gaps of chromatographic resin in the first section, and recycling the water after collection; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and the fourth stage collects high-purity L-malic acid solution, namely L-malic acid separating solution. The TY019 resin is a terpolymer synthesized by taking EA, BA and TYH-SFLP-Buty1 as raw materials.
3) Concentration and crystallization: concentrating the L-malic acid separation liquid at 50 deg.C under reduced pressure until the specific gravity is 1.2, stirring at 40 deg.C for 30min, adding seed crystal, growing crystal for 1h, cooling to 20 deg.C at 3 deg.C/h, then cooling to 5 deg.C at 5 deg.C/h, and centrifuging to separate solid and liquid to obtain wet L-malic acid.
4) Drying: placing the wet L-malic acid in a vacuum drying oven, firstly pumping at normal temperature for 1h by a pump until the water content is below 2%, then heating at a speed of 8 ℃/h in a gradient manner until the temperature is up to 55 ℃, and drying in vacuum for 5h to obtain the finished L-malic acid product with the purity of 99.8%.
Example 3
The method for extracting the L-malic acid comprises the following steps:
1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification until the pH value is 2.5, carrying out vacuum filtration to remove calcium salt, mycelia and fermentation residues, adding deionized water into a filter cake obtained by filtration, washing until the pH value is 5.0, so as to ensure that no malic acid is lost, and obtaining L-malic acid acidification liquor.
2) Removing impurities: feeding the L-malic acid acidized solution into a TY019 separation resin column at the temperature of 45 ℃, wherein the flow rate of the column is 25mL/min, the feeding amount is 200g, collecting the separation solution in sections, collecting water in gaps of chromatographic resin in the first section, and recycling the water; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and the fourth stage collects high-purity L-malic acid solution, namely L-malic acid separating solution. The TY019 resin is a terpolymer synthesized by taking EA, BA and TYH-SFLP-Buty1 as raw materials.
3) Concentration and crystallization: concentrating the L-malic acid separation liquid at 70 ℃ under reduced pressure until the specific gravity is 1.4, stirring at 30 ℃ for 30min, adding seed crystals, growing crystals for 1h, cooling to 20 ℃ at the speed of 3 ℃/h, then cooling to 8 ℃ at the speed of 5 ℃/h in a gradient manner, and carrying out vacuum filtration to separate solid and liquid so as to obtain the wet L-malic acid.
4) Drying: placing the wet L-malic acid in a vacuum drying oven, firstly pumping at normal temperature for 1h by a pump until the water content is below 2%, then heating at a speed of 8 ℃/h in a gradient manner until the temperature is up to 55 ℃, and drying in vacuum for 3h to obtain the finished L-malic acid product with the purity of 99.9%.
Example 4
The method for extracting the L-malic acid comprises the following steps:
1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification until the pH value is 3.0, carrying out vacuum filtration to remove calcium salt, mycelia and fermentation residues, adding deionized water into a filter cake obtained by filtration, and washing until the pH value is 5.0 so as to ensure that no malic acid is lost, thereby obtaining L-malic acid acidification liquor.
2) Removing impurities: feeding the L-malic acid acidized solution into a TY019 separation resin column at the temperature of 25 ℃, wherein the flow rate of the column is 60mL/min, the feeding amount is 200g, collecting the separation solution in sections, collecting water in gaps of chromatographic resin in the first section, and recycling the water; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and the fourth stage collects high-purity L-malic acid solution, namely L-malic acid separating solution. The TY019 resin is a terpolymer synthesized by taking EA, BA and TYH-SFLP-Buty1 as raw materials.
3) Concentration and crystallization: concentrating the L-malic acid separation liquid at 80 deg.C under reduced pressure until the specific gravity is 1.5, stirring at 60 deg.C for 30min, adding seed crystal, growing crystal for 1h, cooling to 20 deg.C at 3 deg.C/h, then cooling to 6 deg.C at 5 deg.C/h, and centrifuging to separate solid and liquid to obtain wet L-malic acid.
4) Drying: placing the wet L-malic acid in a vacuum drying oven, firstly pumping at normal temperature for 1h by a pump until the water content is below 2%, then heating at a speed of 8 ℃/h in a gradient manner until the temperature is up to 55 ℃, and drying in vacuum for 4h to obtain the finished L-malic acid product with the purity of 99.8%.
Example 5
The method for extracting the L-malic acid comprises the following steps:
1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification until the pH value is 1.5, carrying out vacuum filtration to remove calcium salt, mycelia and fermentation residues, adding deionized water into a filter cake obtained by filtration, washing until the pH value is 5.0, so as to ensure that no malic acid is lost, and obtaining L-malic acid acidification liquor.
2) Removing impurities: feeding the L-malic acid acidized solution into a TY019 separation resin column at the temperature of 35 ℃, wherein the flow rate of the column is 50mL/min, the feeding amount is 100g, collecting the separated solution in sections, collecting water in gaps of chromatographic resin in the first section, and recycling the water after collection; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and the fourth stage collects high-purity L-malic acid solution, namely L-malic acid separating solution. The TY019 resin is a terpolymer synthesized by taking EA, BA and TYH-SFLP-Buty1 as raw materials.
3) Concentration and crystallization: concentrating the L-malic acid separation liquid at 40 deg.C under reduced pressure until the specific gravity is 1.3, stirring at 50 deg.C for 30min, adding seed crystal, growing crystal for 1h, cooling to 20 deg.C at 3 deg.C/h, then cooling to 2 deg.C at 5 deg.C/h, and centrifuging to separate solid and liquid to obtain wet L-malic acid.
4) Drying: placing the wet L-malic acid in a vacuum drying oven, firstly pumping at normal temperature for 1h by a pump until the water content is below 2%, then heating at a speed of 8 ℃/h in a gradient manner until the temperature is up to 55 ℃, and drying in vacuum for 3h to obtain the finished L-malic acid product with the purity of 99.7%.
Comparative example 1
The separation resin column was replaced with a DTF-02 resin for chromatography, and the other conditions were the same as in example 1.
Comparative example 2
The separation resin column was replaced with LX-T83SS hydrogen type chromatography resin, and the other conditions were kept the same as in example 1.
Comparative example 3
The separation resin column was replaced with 312 weakly basic anion resin and the other conditions were kept the same as in example 1.
The content of the collected L-malic acid in example 1 and comparative examples 1-3 was measured by high performance liquid chromatography, and the purity of the L-malic acid was calculated from the content of the L-malic acid at the acidity, and the yield of the L-malic acid was further calculated. The results are shown in Table 1.
TABLE 1 purity and yield of L-malic acid obtained in example 1 and comparative examples 1-3
As can be seen from Table 1, the purity of the L-malic acid obtained from the TY-019 type resin can reach more than 98%, and the yield is higher than that of other resins.
Claims (8)
- The method for extracting the L-malic acid is characterized by comprising the following steps of:1) acidifying: adding oxalic acid into the L-malic acid fermentation liquor for acidification, and filtering to obtain a filtrate, namely an L-malic acid acidification liquor;2) separating and removing impurities: passing the L-malic acid acidified liquid obtained in the step 1) through a separation resin column to obtain an L-malic acid separation liquid;3) concentration and crystallization: concentrating and crystallizing the L-malic acid separation liquid obtained in the step 2) to obtain an L-malic acid wet product;4) gradient drying: and (3) drying the wet L-malic acid product in a vacuum gradient manner to obtain a finished L-malic acid product.
- 2. The method for extracting L-malic acid according to claim 1, wherein oxalic acid is added to the fermentation broth of L-malic acid in step 1) to acidify the fermentation broth to pH 1.5-3.0.
- 3. The method for extracting L-malic acid as claimed in claim 1, wherein the separating resin column in step 2) is TY019 separating resin column, and TY019 resin is terpolymer synthesized from EA, BA and TYH-SFLP-Buty 1.
- 4. The method for extracting L-malic acid according to claim 1 or 3, wherein, when the resin column is separated in step 2), the separated liquid is collected in stages, and the water in the gaps of the resin column is collected and separated in the first stage for recycling; collecting impurity solution in the second section, and discarding; collecting low-purity L-malic acid solution in the third stage, and storing for next separation; and collecting the high-purity L-malic acid solution in the fourth stage, namely the L-malic acid separation solution.
- 5. The method for extracting L-malic acid according to claim 1 or 4, wherein the concentration in step 2) is carried out by concentrating the L-malic acid separated liquid at 40-80 ℃ under reduced pressure.
- 6. The method for extracting L-malic acid according to claim 1, wherein the concentration in step 2) is to concentrate the L-malic acid separated liquid to a specific gravity of 1.2 to 1.5.
- 7. The method for extracting L-malic acid according to claim 1, wherein the crystallization in step 3) is performed by stirring at 30-60 ℃, adding seed crystals, growing crystals for 1h, cooling to 20 ℃ at a rate of 3 ℃/h, then cooling to 0-10 ℃ at a rate of 5 ℃/h in a gradient manner, and vacuum filtering or centrifugal separation of solid and liquid.
- 8. The method for extracting L-malic acid according to claim 1, wherein the gradient drying in step 4) is performed by cold-pumping the wet L-malic acid at room temperature until the water content is below 2%, then heating at a rate of 8 ℃/h with a gradient of up to 55 ℃, drying for 3-5h, and sieving to obtain the final product.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114717009A (en) * | 2022-04-29 | 2022-07-08 | 湖州中恒园林建设有限公司 | Organic acid sustained-release preparation for improving saline-alkali soil and preparation method thereof |
| CN114989009A (en) * | 2022-06-29 | 2022-09-02 | 安徽雪郎生物科技股份有限公司 | Preparation method of anti-caking DL-malic acid |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102746348A (en) * | 2011-04-19 | 2012-10-24 | 上海医药工业研究院 | Method for separation of lincomycin |
| CN103642853A (en) * | 2013-12-02 | 2014-03-19 | 山东阜丰发酵有限公司 | Novel process for extracting L-malic acid |
| WO2016124170A1 (en) * | 2015-02-06 | 2016-08-11 | Rwth Aachen | Method for isolating organic dicarboxlic acids by adsorption on hydrophobic porous materials |
| CN110256227A (en) * | 2019-07-02 | 2019-09-20 | 安徽丰原发酵技术工程研究有限公司 | A method of extracting production malic acid from calcium malate solution |
-
2020
- 2020-08-24 CN CN202010859506.3A patent/CN111892498A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102746348A (en) * | 2011-04-19 | 2012-10-24 | 上海医药工业研究院 | Method for separation of lincomycin |
| CN103642853A (en) * | 2013-12-02 | 2014-03-19 | 山东阜丰发酵有限公司 | Novel process for extracting L-malic acid |
| WO2016124170A1 (en) * | 2015-02-06 | 2016-08-11 | Rwth Aachen | Method for isolating organic dicarboxlic acids by adsorption on hydrophobic porous materials |
| CN110256227A (en) * | 2019-07-02 | 2019-09-20 | 安徽丰原发酵技术工程研究有限公司 | A method of extracting production malic acid from calcium malate solution |
Non-Patent Citations (2)
| Title |
|---|
| 何领好主编: "《功能高分子材料》", 31 August 2016, 华中科技大学出版社 * |
| 陈淑伟主编, 哈尔滨工程大学出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114717009A (en) * | 2022-04-29 | 2022-07-08 | 湖州中恒园林建设有限公司 | Organic acid sustained-release preparation for improving saline-alkali soil and preparation method thereof |
| CN114989009A (en) * | 2022-06-29 | 2022-09-02 | 安徽雪郎生物科技股份有限公司 | Preparation method of anti-caking DL-malic acid |
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