CN113564134A - Process for extracting alliinase effective component from garlic - Google Patents
Process for extracting alliinase effective component from garlic Download PDFInfo
- Publication number
- CN113564134A CN113564134A CN202110640266.2A CN202110640266A CN113564134A CN 113564134 A CN113564134 A CN 113564134A CN 202110640266 A CN202110640266 A CN 202110640266A CN 113564134 A CN113564134 A CN 113564134A
- Authority
- CN
- China
- Prior art keywords
- alliinase
- garlic
- extracting
- active ingredients
- meshes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010092760 Alliin lyase Proteins 0.000 title claims abstract description 138
- 235000004611 garlic Nutrition 0.000 title claims abstract description 103
- 238000000034 method Methods 0.000 title claims abstract description 48
- 244000245420 ail Species 0.000 title 1
- 240000002234 Allium sativum Species 0.000 claims abstract description 102
- 238000000605 extraction Methods 0.000 claims abstract description 31
- 238000001914 filtration Methods 0.000 claims abstract description 19
- 210000004911 serous fluid Anatomy 0.000 claims abstract description 19
- 238000004140 cleaning Methods 0.000 claims abstract description 16
- 238000001035 drying Methods 0.000 claims abstract description 10
- 239000011268 mixed slurry Substances 0.000 claims abstract description 10
- 238000004506 ultrasonic cleaning Methods 0.000 claims abstract description 9
- 239000002351 wastewater Substances 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims description 33
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 22
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 22
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 22
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 18
- 239000004480 active ingredient Substances 0.000 claims description 17
- 239000002244 precipitate Substances 0.000 claims description 16
- 108090000790 Enzymes Proteins 0.000 claims description 15
- 102000004190 Enzymes Human genes 0.000 claims description 15
- 238000000502 dialysis Methods 0.000 claims description 14
- 235000011837 pasties Nutrition 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 239000002002 slurry Substances 0.000 claims description 12
- 239000003963 antioxidant agent Substances 0.000 claims description 9
- 230000003078 antioxidant effect Effects 0.000 claims description 9
- 239000006172 buffering agent Substances 0.000 claims description 9
- 239000008139 complexing agent Substances 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 8
- 230000001276 controlling effect Effects 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000011261 inert gas Substances 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 238000007873 sieving Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 4
- 238000011049 filling Methods 0.000 claims description 4
- 239000005457 ice water Substances 0.000 claims description 4
- 229920002635 polyurethane Polymers 0.000 claims description 4
- 239000004814 polyurethane Substances 0.000 claims description 4
- AEUVIXACNOXTBX-UHFFFAOYSA-N 1-sulfanylpropan-1-ol Chemical group CCC(O)S AEUVIXACNOXTBX-UHFFFAOYSA-N 0.000 claims description 3
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 3
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical group [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 3
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 238000000354 decomposition reaction Methods 0.000 abstract description 10
- 238000012216 screening Methods 0.000 abstract description 10
- 239000012535 impurity Substances 0.000 abstract description 7
- 230000036632 reaction speed Effects 0.000 abstract description 6
- 238000000227 grinding Methods 0.000 abstract description 4
- 239000000243 solution Substances 0.000 description 11
- XUHLIQGRKRUKPH-GCXOYZPQSA-N Alliin Natural products N[C@H](C[S@@](=O)CC=C)C(O)=O XUHLIQGRKRUKPH-GCXOYZPQSA-N 0.000 description 8
- XUHLIQGRKRUKPH-UHFFFAOYSA-N S-allyl-L-cysteine sulfoxide Natural products OC(=O)C(N)CS(=O)CC=C XUHLIQGRKRUKPH-UHFFFAOYSA-N 0.000 description 8
- XUHLIQGRKRUKPH-DYEAUMGKSA-N alliin Chemical compound OC(=O)[C@@H](N)C[S@@](=O)CC=C XUHLIQGRKRUKPH-DYEAUMGKSA-N 0.000 description 8
- 235000015295 alliin Nutrition 0.000 description 8
- 239000012530 fluid Substances 0.000 description 7
- 238000000926 separation method Methods 0.000 description 6
- 238000003860 storage Methods 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010612 desalination reaction Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 description 2
- IXELFRRANAOWSF-FNORWQNLSA-N (E)-Ajoene Chemical compound C=CCSS\C=C\CS(=O)CC=C IXELFRRANAOWSF-FNORWQNLSA-N 0.000 description 1
- JDLKFOPOAOFWQN-VIFPVBQESA-N Allicin Natural products C=CCS[S@](=O)CC=C JDLKFOPOAOFWQN-VIFPVBQESA-N 0.000 description 1
- 241000234282 Allium Species 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000234280 Liliaceae Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- IXELFRRANAOWSF-CYBMUJFWSA-N ajoene Natural products C=CCSSC=CC[S@](=O)CC=C IXELFRRANAOWSF-CYBMUJFWSA-N 0.000 description 1
- -1 alkyl thiocysteine Chemical compound 0.000 description 1
- JDLKFOPOAOFWQN-UHFFFAOYSA-N allicin Chemical compound C=CCSS(=O)CC=C JDLKFOPOAOFWQN-UHFFFAOYSA-N 0.000 description 1
- 235000010081 allicin Nutrition 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- IXELFRRANAOWSF-UHFFFAOYSA-N cis-ajoene Natural products C=CCSSC=CCS(=O)CC=C IXELFRRANAOWSF-UHFFFAOYSA-N 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000004537 pulping Methods 0.000 description 1
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 description 1
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 description 1
- 229960001327 pyridoxal phosphate Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a process method for extracting allinase effective components from garlic, which relates to the technical field of allinase extraction processes and comprises the following steps: cleaning garlic, preparing crude alliinase serous fluid, dialyzing the alliinase serous fluid, drying to prepare alliinase extract, screening the garlic before cleaning, removing the garlic which is damaged by worms, rotten or mildewed, cleaning the garlic by adopting ultrasonic cleaning equipment, and purifying the cleaning wastewater. The invention adopts the matching of garlic screening, cleaning and grinding and filtering, removes the impurity influence in the garlic by screening and cleaning the garlic, ensures the purity of the garlic during extraction, then slices and grinds the garlic to form the filtration of the garlic mixed slurry, and slows down the decomposition reaction speed of the alliinase by matching with the filtration within the temperature range of-5 ℃ to 5 ℃, thereby avoiding the inconvenience of extracting the alliinase due to the higher decomposition reaction speed of the alliinase under the conventional environment, improving the extraction efficiency of the alliinase and reducing the extraction cost.
Description
Technical Field
The invention relates to the technical field of allinase extraction processes, in particular to a process method for extracting allinase effective components from garlic.
Background
The garlic, also called the garlic, is the underground bulb of the garlic of the genus allium of the family liliaceae, recent research finds that the garlic has the effects of resisting cancer, reducing blood fat, killing fungi and the like, the alliinase, also called alliin lyase, alkyl thiocysteine enzyme, exists in the vacuole of the cell, consists of six pyridoxal phosphate and two subunits, has the maximum absorption at the 280 nm and 430nm, and at present, the alliinase is generally accepted to catalyze alliin to generate a series of volatile sulfur-containing organic compounds such as allicin, ajoene and the like to play a plurality of drug effects, so the use of the alliinase plays an important role in the aspect of the prescription, and the process for extracting the active ingredients of the alliinase from the garlic is particularly important.
The following problems exist in the prior art:
1. the existing extraction of effective components of allinase in garlic has the problems of low extraction efficiency and high extraction cost of allinase in garlic due to the inconvenience of extraction caused by the high decomposition reaction speed of allinase under the conventional environment;
2. when the alliinase in the garlic is extracted, the adhesion degree of the alliinase and alliin is higher, so that the extraction difficulty of the alliinase is further increased, the extraction yield of the alliinase is reduced, and the extraction workload is increased.
Disclosure of Invention
The invention provides a process method for extracting alliinase active ingredients from garlic, which aims to have the capability of slowing down alliinase decomposition reaction and solve the problem that the alliinase is inconvenient to extract because the alliinase decomposition reaction is faster under the conventional environment; the other purpose is to solve the problem that the extraction difficulty of the alliinase is further increased due to higher adhesion degree of the alliinase and alliin when the alliinase in the garlic is extracted, so that the effects of improving the extraction yield of the alliinase and reducing the extraction workload are achieved.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a process method for extracting alliinase active ingredients from garlic comprises the following steps:
step one, cleaning garlic;
step two, preparing coarse alliinase serous fluid;
step three, performing alliinase serous fluid dialysis;
and step four, drying to prepare the allinase extract.
The technical scheme of the invention is further improved as follows: the first step further comprises: the garlic is screened before being cleaned, the garlic which is damaged by worms, rotten or mildewed is removed, the cleaned garlic can be cleaned by adopting ultrasonic cleaning equipment, and cleaning wastewater needs to be purified, wherein the model of the ultrasonic cleaning equipment is JP-100 PLUS.
By adopting the technical scheme, the garlic in the scheme needs to be screened and cleaned by adopting ultrasonic cleaning equipment before being cleaned, so that the purity of the garlic allinase is greatly improved during extraction.
The technical scheme of the invention is further improved as follows: the second step further comprises: before the coarse alliinase serous fluid is prepared, clean garlic needs to be sliced and ground to form garlic mixed serous fluid, and the garlic mixed serous fluid needs to be filtered for multiple times by adopting filter screens with different meshes.
The technical scheme of the invention is further improved as follows: the filter screens are respectively 500 meshes, 400 meshes and 200 meshes of 450 meshes, 300 meshes and 400 meshes and 100 meshes, the filtering temperature of the garlic mixed slurry is controlled within the range of-5 ℃ to 5 ℃, and the crude garlic enzyme slurry can be obtained after the garlic mixed slurry is separated.
By adopting the technical scheme, the temperature of-5 ℃ to 5 ℃ and the filtering of the filter screens with different meshes are adopted in the scheme, so that the decomposition reaction speed of the alliinase is effectively slowed down, and the separation rate of the alliinase is improved.
The technical scheme of the invention is further improved as follows: the third step further comprises the following steps:
A. filtering the alliinase slurry;
B. sieving the clear and turbid liquid for multiple times;
C. adding medium to concentrate the turbid solution.
The technical scheme of the invention is further improved as follows: the B also comprises: keeping the temperature range of-5 ℃ to 5 ℃, sieving the clear solution generated by filtering the alliinase slurry for 3-4 times, fully separating the alliinase in the clear solution to obtain a first clear solution of the alliinase, wherein the step C further comprises the following steps: stirring the first turbid liquid of the alliinase, adding a medium into the first turbid liquid of the alliinase, adding ammonium sulfate into the first turbid liquid of the alliinase to completely dissolve the first turbid liquid of the alliinase, controlling the concentration of the ammonium sulfate to be within the range of 20 to 30 percent by weight and volume percent by a hydrometer, separating to obtain a second turbid liquid of the alliinase, and separating by adopting a freezing closed type pulp residue self-separation pulp grinder or a centrifugal drying machine or a low-temperature tubular centrifugal separator.
By adopting the technical scheme, the first turbid liquid of the alliinase and the second turbid liquid of the alliinase are separated and prepared, so that the mutual influence between the alliinase and alliin is reduced, and the extraction purity of the alliinase is guaranteed.
The technical scheme of the invention is further improved as follows: and stirring the second clear liquid of the alliinase at the temperature of between 5 ℃ below zero and 5 ℃ and adding ammonium sulfate into the second clear liquid of the alliinase to completely dissolve the second clear liquid of the alliinase, controlling the concentration of the ammonium sulfate to be between 40 and 50 percent by weight volume percent by using a hydrometer, and separating out crude alliinase of a first pasty precipitate.
The technical scheme of the invention is further improved as follows: the medium comprises ammonium sulfate, a complexing agent, an antioxidant, a buffering agent and a water solution of glycerol, wherein the complexing agent is disodium ethylene diamine tetraacetate, the antioxidant is mercaptopropanol, and the buffering agent is phosphate.
By adopting the technical scheme, the ammonium, the complexing agent, the antioxidant, the buffering agent and the aqueous solution of the glycerol are matched together, so that the separation efficiency and the separation purity of the allinase are improved.
The technical scheme of the invention is further improved as follows: the fourth step further comprises: preparing alliinase extract by adopting the processes of dialysis desalination-isoelectric point regulation-freeze drying, filling crude alliinase of the first pasty precipitate into a polyurethane bag, dialyzing for 40 to 50 hours in a flowing ice water dialysis tank at the temperature of between 5 ℃ below zero and 5 ℃, and separating out second pasty precipitate after the pH value of the isoelectric point is regulated to 4.9 by using phosphoric acid.
By adopting the technical scheme, the process of dialysis desalination, isoelectric point adjustment and freeze drying in the scheme ensures the extraction yield of alliinase extraction and reduces the workload of extraction.
The technical scheme of the invention is further improved as follows: and (3) putting the second pasty precipitate into a freeze dryer for drying until the water content is lower than 2.0% to obtain freeze-dried alliinase, and hermetically storing with nitrogen or inert gas, wherein the model of the freeze dryer is FD-1B-50.
By adopting the technical scheme, the nitrogen or inert gas in the scheme is used for hermetically storing the freeze-dried alliinase, so that the stability of the storage environment of the alliinase extract is ensured.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the technical progress that:
1. the invention provides a process for extracting active ingredients of alliinase from garlic, which adopts the matching of garlic screening, cleaning and grinding and filtering, removes the influence of impurities in the garlic by screening and cleaning the garlic, ensures the purity of the garlic during extraction, then forms filtration after garlic mixed slurry by slicing and grinding, slows down the decomposition reaction speed of the alliinase by matching with the filtration within the temperature range of-5 ℃ to 5 ℃, avoids the inconvenience of extracting the alliinase due to higher decomposition reaction speed of the alliinase under the conventional environment, improves the extraction efficiency of the alliinase and reduces the extraction cost.
2. The invention provides a process method for extracting alliinase active ingredients from garlic, which adopts the cooperation of screening garlic enzyme clear liquid for multiple times and adding a medium to concentrate the clear liquid, fully separates impurities in the clear liquid by filtering the garlic enzyme serous fluid for multiple times, reduces the mutual influence between alliinase and alliin by utilizing the addition of the medium such as ammonium sulfate, a complexing agent, an antioxidant, a buffering agent, glycerol aqueous solution and the like, improves the purity of the alliinase, avoids the problem that the extraction difficulty of the alliinase is further increased due to higher adhesion of the alliinase and the alliin when the alliinase in the garlic is extracted, improves the extraction yield of the alliinase, and reduces the workload when the alliinase is extracted.
3. The invention provides a process method for extracting alliinase active ingredients from garlic, which adopts the procedures of dialysis desalination, isoelectric point adjustment and freeze drying to prepare an alliinase extract, further improves the extraction and preparation efficiency of alliinase, and ensures the stability of the environment during the storage of the alliinase by hermetically storing the freeze-dried alliinase by nitrogen or inert gas, thereby furthest ensuring the storage and utilization efficiency of the alliinase.
Drawings
FIG. 1 is a flow chart of the present invention;
FIG. 2 is a schematic diagram of the garlic enzyme serous fluid dialysis process of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples:
example 1
As shown in FIGS. 1-2, the present invention provides a process for extracting alliinase active ingredients from garlic, which comprises the following steps:
step one, garlic cleaning, which specifically comprises the following steps: the garlic is screened before being cleaned, the garlic which is damaged by worms, rotten or mildewed is removed, the influence of impurities in the garlic on the subsequent process is removed by screening the garlic, the garlic can be cleaned by adopting ultrasonic cleaning equipment, the garlic is sufficiently cleaned by matching with the cleaning of the ultrasonic cleaning equipment, the impurities stained on the garlic are removed, the cleaning wastewater needs to be purified, and the model of the ultrasonic cleaning equipment is JP-100 PLUS;
step two, preparing coarse alliinase slurry, which specifically comprises the following steps: before the preparation of the crude garlic enzyme slurry, clean garlic needs to be sliced and ground to form garlic mixed slurry, the garlic mixed slurry needs to be filtered for many times by adopting filter screens with different meshes, the filter screens are respectively 500 meshes, 300 meshes and 400 meshes and 100 meshes and 200 meshes, the filtering temperature of the garlic mixed slurry is controlled within the range of-5 ℃ to 5 ℃, the garlic slurry is filtered by utilizing the temperature range of-5 ℃ to 5 ℃, the cell wall damage after the garlic is crushed is relieved, the decomposition efficiency of the garlic enzyme and alliin is relieved, the extraction efficiency of the garlic enzyme is further improved, and the crude garlic enzyme slurry can be obtained after the separation of the garlic mixed slurry;
step three, performing alliinase serous fluid dialysis;
and step four, drying to prepare the allinase extract.
Example 2
As shown in fig. 1-2, on the basis of embodiment 1, the present invention provides a technical solution: preferably, step three further comprises the steps of:
A. filtering the alliinase slurry;
B. screening the clear and turbid liquid for multiple times, which specifically comprises the following steps: keeping the temperature range of-5 ℃ to 5 ℃, sieving the clear solution generated by filtering the alliinase slurry for 3-4 times, and fully separating the alliinase in the clear solution from impurities to obtain a first clear solution of the alliinase;
C. adding a medium to concentrate the turbid solution, which specifically comprises the following steps: stirring the first turbid liquid of the alliinase, adding a medium into the first turbid liquid of the alliinase, adding ammonium sulfate into the first turbid liquid of the alliinase to completely dissolve the ammonium sulfate, controlling the concentration of the ammonium sulfate to be within the range of 20 to 30 weight percent by volume by a gravimeter, separating to obtain a second turbid liquid of the alliinase, separating by adopting a freezing closed type pulp residue self-separation pulping machine or a centrifugal drying machine or a low-temperature tubular centrifugal separator, stirring and adding the ammonium sulfate into the second turbid liquid of the alliinase to completely dissolve the second turbid liquid of the alliinase at the temperature of between 5 ℃ below zero and 5 ℃, controlling the concentration of the ammonium sulfate to be within the range of 40 to 50 weight percent by volume by the gravimeter, separating out crude alliinase of a first pasty precipitate, wherein the medium comprises an aqueous solution of ammonium sulfate, a complexing agent, an antioxidant, a buffering agent and glycerol, the complexing agent is disodium ethylene diamine tetraacetate, the antioxidant is mercaptopropanol, the buffering agent is phosphate, by adding the ammonium sulfate, the complexing agent, the antioxidant, the buffering agent, the glycerol aqueous solution and other media, the decomposition reaction of the alliinase and the alliin is further slowed down, the purity of the alliinase is improved, and the yield of the alliinase in preparation is guaranteed.
Example 3
As shown in fig. 1-2, on the basis of embodiment 1 and embodiment 2, the present invention provides a technical solution: preferably, the fourth step further comprises: preparing alliinase extract by adopting a dialysis desalting-isoelectric point adjusting-freeze drying process, reducing the preparation difficulty of alliinase, filling crude alliinase of a first paste precipitate into a polyurethane bag, dialyzing in a flowing ice water dialysis tank at the temperature of between 5 ℃ below zero and 5 ℃ for 40 to 50 hours, adjusting the pH of the isoelectric point to 4.9 by using phosphoric acid, separating a second paste precipitate, drying the second paste precipitate in a freeze dryer to ensure that the water content is lower than 2.0 percent to obtain the alliinase, performing closed storage by using nitrogen or inert gas to ensure the storage efficiency of the alliinase to the maximum extent, wherein the model of the freeze dryer is FD-1B-50, and meanwhile, the alliinase product is proved to have higher purity by SDS-gel electrophoresis, and the molecular weight of a homopolymer of the alliinase product is 67000 daltons according to the specific transfer value of standard protein (C molecular weight 14400 to 97400), the Km of the Michaelis constant is 3.6mmol/L, and the average activity of the product is 765U/g.
As shown in fig. 1-2, when extracting the active components of alliinase in garlic, firstly screening garlic to remove garlic damaged by worms, rotten or mildewed garlic, then cleaning the garlic by using ultrasonic cleaning equipment, slicing and grinding clean garlic to prepare garlic mixed serous fluid, then filtering the garlic mixed serous fluid for a plurality of times by using filter screens with different meshes within the temperature range of-5 ℃ to 5 ℃, dialyzing crude alliinase serous fluid, screening the clear fluid generated by filtering the alliinase serous fluid for 3-4 times within the temperature range of-5 ℃ to 5 ℃, fully separating alliinase in the clear fluid from impurities to obtain first garlic enzyme clear fluid, stirring the first garlic enzyme clear fluid, adding a medium into the first garlic enzyme clear fluid, adding ammonium sulfate into the first garlic enzyme clear fluid to completely dissolve the garlic enzyme clear fluid, and controlling the concentration of the ammonium sulfate within the range of 20% to 30% by weight volume percentage by using a gravimeter, separating to obtain second turbid liquid of alliinase, stirring the second turbid liquid of alliinase, adding ammonium sulfate to completely dissolve the second turbid liquid of alliinase, controlling the concentration of the ammonium sulfate to be in a range of 40-50% by weight volume percent by using a gravimeter, separating out coarse alliinase of a first pasty precipitate, preparing an alliinase extract by adopting a process of dialysis desalting-isoelectric point adjusting-freeze drying, filling the coarse alliinase of the first pasty precipitate into a polyurethane bag, dialyzing in a flowing ice water dialysis tank at a temperature of-5 ℃ for 40-50 h, adjusting the pH of the isoelectric point to 4.9 by using phosphoric acid, separating out a second pasty precipitate, putting the second pasty precipitate into a freeze dryer for drying to ensure that the moisture content is lower than 2.0 percent to obtain the alliinase, and hermetically storing by using nitrogen or inert gas.
The present invention has been described in general terms in the foregoing, but it will be apparent to those skilled in the art that modifications and improvements can be made thereto based on the present invention. Therefore, modifications or improvements are within the scope of the invention without departing from the spirit of the inventive concept.
Claims (10)
1. A process method for extracting alliinase active ingredients from garlic is characterized by comprising the following steps: the process method for extracting the allinase effective component from the garlic comprises the following steps:
step one, cleaning garlic;
step two, preparing coarse alliinase serous fluid;
step three, performing alliinase serous fluid dialysis;
and step four, drying to prepare the allinase extract.
2. The process method for extracting alliinase active ingredients from garlic as claimed in claim 1, wherein the process comprises the steps of: the first step further comprises: the garlic is screened before being cleaned, the garlic which is damaged by worms, rotten or mildewed is removed, the cleaned garlic can be cleaned by ultrasonic cleaning equipment, and cleaning wastewater needs to be purified.
3. The process method for extracting alliinase active ingredients from garlic as claimed in claim 1, wherein the process comprises the steps of: the second step further comprises: before the coarse alliinase serous fluid is prepared, clean garlic needs to be sliced and ground to form garlic mixed serous fluid, and the garlic mixed serous fluid needs to be filtered for multiple times by adopting filter screens with different meshes.
4. The process method for extracting alliinase active ingredients from garlic as claimed in claim 3, wherein the process comprises the steps of: the filter screens are respectively 500 meshes, 400 meshes and 200 meshes of 450 meshes, 300 meshes and 400 meshes and 100 meshes, the filtering temperature of the garlic mixed slurry is controlled within the range of-5 ℃ to 5 ℃, and the crude garlic enzyme slurry can be obtained after the garlic mixed slurry is separated.
5. The process method for extracting alliinase active ingredients from garlic as claimed in claim 1, wherein the process comprises the steps of: the third step further comprises the following steps:
A. filtering the alliinase slurry;
B. sieving the clear and turbid liquid for multiple times;
C. adding medium to concentrate the turbid solution.
6. The process of claim 5, wherein the extraction of alliinase-derived active ingredients from garlic comprises: the B also comprises: keeping the temperature range of-5 ℃ to 5 ℃, sieving the clear solution generated by filtering the alliinase slurry for 3-4 times, fully separating the alliinase in the clear solution to obtain a first clear solution of the alliinase, wherein the step C further comprises the following steps: stirring the first turbid liquid of the alliinase, adding a medium into the first turbid liquid of the alliinase, adding ammonium sulfate into the first turbid liquid of the alliinase to completely dissolve the first turbid liquid of the alliinase, controlling the concentration of the ammonium sulfate to be within the range of 20 to 30 weight percent by volume by using a hydrometer, and separating to obtain a second turbid liquid of the alliinase.
7. The process of claim 6, wherein the extraction of alliinase-derived active ingredients from garlic comprises: and stirring the second clear liquid of the alliinase at the temperature of between 5 ℃ below zero and 5 ℃ and adding ammonium sulfate into the second clear liquid of the alliinase to completely dissolve the second clear liquid of the alliinase, controlling the concentration of the ammonium sulfate to be between 40 and 50 percent by weight volume percent by using a hydrometer, and separating out crude alliinase of a first pasty precipitate.
8. The process of claim 5, wherein the extraction of alliinase-derived active ingredients from garlic comprises: the medium comprises ammonium sulfate, a complexing agent, an antioxidant, a buffering agent and a water solution of glycerol, wherein the complexing agent is disodium ethylene diamine tetraacetate, the antioxidant is mercaptopropanol, and the buffering agent is phosphate.
9. The process method for extracting alliinase active ingredients from garlic as claimed in claim 1, wherein the process comprises the steps of: the fourth step further comprises: preparing alliinase extract by adopting the processes of dialysis desalination-isoelectric point regulation-freeze drying, filling crude alliinase of the first pasty precipitate into a polyurethane bag, dialyzing for 40 to 50 hours in a flowing ice water dialysis tank at the temperature of between 5 ℃ below zero and 5 ℃, and separating out second pasty precipitate after the pH value of the isoelectric point is regulated to 4.9 by using phosphoric acid.
10. The process of claim 9, wherein the extraction of alliinase-derived active ingredients from garlic comprises: and (3) putting the second pasty precipitate into a freeze dryer for drying until the water content is lower than 2.0 percent to obtain freeze-dried alliinase, and hermetically storing with nitrogen or inert gas.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110640266.2A CN113564134A (en) | 2021-06-08 | 2021-06-08 | Process for extracting alliinase effective component from garlic |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110640266.2A CN113564134A (en) | 2021-06-08 | 2021-06-08 | Process for extracting alliinase effective component from garlic |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113564134A true CN113564134A (en) | 2021-10-29 |
Family
ID=78161840
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110640266.2A Pending CN113564134A (en) | 2021-06-08 | 2021-06-08 | Process for extracting alliinase effective component from garlic |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113564134A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1424397A (en) * | 2003-01-13 | 2003-06-18 | 陈坚 | Producing process for extracting allinase from garlic |
-
2021
- 2021-06-08 CN CN202110640266.2A patent/CN113564134A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1424397A (en) * | 2003-01-13 | 2003-06-18 | 陈坚 | Producing process for extracting allinase from garlic |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101319208B (en) | Method for preparation of bromelin | |
| CN108299520A (en) | Promote the method and aurantiamarin of extraction aurantiamarin purity | |
| CN104277915A (en) | Method for extracting peony essential oil from peony | |
| CN107011992A (en) | A kind of method of molecular clock Fructus Fici extract | |
| CN104744611A (en) | Collaborative cleaning extraction method for active ingredients of Taraxacum kok-saghyz Rodin | |
| CN106008211A (en) | Method for extraction of cynarin from cynara scolymus by membrane integration technology | |
| CN102660137A (en) | Method for high-efficiency preparation of natural pigment of Ziziphus jujuba Mill. | |
| CN104231097A (en) | Preparation method of lyceum barbarum polysaccharide | |
| CN1091152C (en) | Physical extraction of bromelain | |
| CN111471732A (en) | Novel selenium-rich tea source ACE inhibitory peptide and preparation method thereof | |
| CN104844721B (en) | Extraction and separation method of Agrocybe aegirit polysaccharides | |
| CN113564134A (en) | Process for extracting alliinase effective component from garlic | |
| CN110563856A (en) | extraction method of flammulina velutipes polysaccharide | |
| CN102382181B (en) | Method for extracting active proteins with antioxidant and hypoglycemic functions from mushrooms | |
| CN107286268A (en) | A kind of peach gum polysaccharide extracting method | |
| EP3668885B1 (en) | Method of purifying phycocyanin | |
| JP4500078B2 (en) | Extract method of persimmon tannin and persimmon tannin extracted by this method | |
| CN105820237A (en) | Method for two aqueous phase extraction separation of phycocyanin through dextran and sodium potassium tartrate | |
| CN102168120A (en) | Method for improving water solubility of grape seed proanthocyanidin | |
| CN102652556B (en) | Preparation method of instant kudzuvine root particles | |
| CN101805269A (en) | Method for separating and extracting natural theanine | |
| CN109879943B (en) | Extraction method of phycoerythrin | |
| CN114939084A (en) | Russule extract and preparation method and application thereof | |
| EP0218770A1 (en) | Use of marine algae derivatives as culturing agents, as growth factors and as anti-stress agents for microorganisms and fungi, and uses in the coating of seeds and fertilizers | |
| CN109443872A (en) | A kind of raw long-life noodles brown stain product purification methods |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |