CN101760497A - Microorganisms or/and biological enzyme extracting method for plant anthocyanin - Google Patents
Microorganisms or/and biological enzyme extracting method for plant anthocyanin Download PDFInfo
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- CN101760497A CN101760497A CN200910185829A CN200910185829A CN101760497A CN 101760497 A CN101760497 A CN 101760497A CN 200910185829 A CN200910185829 A CN 200910185829A CN 200910185829 A CN200910185829 A CN 200910185829A CN 101760497 A CN101760497 A CN 101760497A
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- anthocyanidin
- anthocyanin
- positively charged
- charged ion
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- 244000005700 microbiome Species 0.000 title claims abstract description 30
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 23
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 16
- 229930002877 anthocyanin Natural products 0.000 title abstract 10
- 235000010208 anthocyanin Nutrition 0.000 title abstract 10
- 239000004410 anthocyanin Substances 0.000 title abstract 10
- 150000004636 anthocyanins Chemical class 0.000 title abstract 8
- -1 Cation anthocyanin Chemical class 0.000 claims abstract description 37
- 238000001556 precipitation Methods 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000006243 chemical reaction Methods 0.000 claims abstract description 14
- 238000005336 cracking Methods 0.000 claims abstract description 10
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 9
- 238000001704 evaporation Methods 0.000 claims abstract description 5
- 238000001914 filtration Methods 0.000 claims abstract description 5
- 230000008014 freezing Effects 0.000 claims abstract description 5
- 238000007710 freezing Methods 0.000 claims abstract description 5
- 238000010438 heat treatment Methods 0.000 claims abstract description 5
- 229930014669 anthocyanidin Natural products 0.000 claims description 62
- 235000008758 anthocyanidins Nutrition 0.000 claims description 62
- 150000001452 anthocyanidin derivatives Chemical class 0.000 claims description 27
- 241000196324 Embryophyta Species 0.000 claims description 20
- 238000003756 stirring Methods 0.000 claims description 20
- 238000006731 degradation reaction Methods 0.000 claims description 18
- 230000015556 catabolic process Effects 0.000 claims description 16
- 238000001035 drying Methods 0.000 claims description 12
- 241000894006 Bacteria Species 0.000 claims description 10
- 239000003945 anionic surfactant Substances 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 102000006995 beta-Glucosidase Human genes 0.000 claims description 8
- 108010047754 beta-Glucosidase Proteins 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 8
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 8
- 102000004169 proteins and genes Human genes 0.000 claims description 8
- 108090000623 proteins and genes Proteins 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 6
- 229920002472 Starch Polymers 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims description 5
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 4
- 101000874334 Dalbergia nigrescens Isoflavonoid 7-O-beta-apiosyl-glucoside beta-glycosidase Proteins 0.000 claims description 4
- 101000757733 Enterococcus faecalis (strain ATCC 700802 / V583) Autolysin Proteins 0.000 claims description 4
- 235000000177 Indigofera tinctoria Nutrition 0.000 claims description 4
- 101000757734 Mycolicibacterium phlei 38 kDa autolysin Proteins 0.000 claims description 4
- 108010029182 Pectin lyase Proteins 0.000 claims description 4
- 235000009392 Vitis Nutrition 0.000 claims description 4
- 241000219095 Vitis Species 0.000 claims description 4
- BTBJBAZGXNKLQC-UHFFFAOYSA-N ammonium lauryl sulfate Chemical compound [NH4+].CCCCCCCCCCCCOS([O-])(=O)=O BTBJBAZGXNKLQC-UHFFFAOYSA-N 0.000 claims description 4
- 229940063953 ammonium lauryl sulfate Drugs 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 108010089934 carbohydrase Proteins 0.000 claims description 4
- 150000001768 cations Chemical class 0.000 claims description 4
- 210000002421 cell wall Anatomy 0.000 claims description 4
- 230000006378 damage Effects 0.000 claims description 4
- 230000006866 deterioration Effects 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 235000013399 edible fruits Nutrition 0.000 claims description 4
- 230000000415 inactivating effect Effects 0.000 claims description 4
- 229940097275 indigo Drugs 0.000 claims description 4
- COHYTHOBJLSHDF-UHFFFAOYSA-N indigo powder Natural products N1C2=CC=CC=C2C(=O)C1=C1C(=O)C2=CC=CC=C2N1 COHYTHOBJLSHDF-UHFFFAOYSA-N 0.000 claims description 4
- 235000014655 lactic acid Nutrition 0.000 claims description 4
- 230000002906 microbiologic effect Effects 0.000 claims description 4
- 238000002203 pretreatment Methods 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 239000000376 reactant Substances 0.000 claims description 4
- 238000011084 recovery Methods 0.000 claims description 4
- 238000004064 recycling Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 239000008234 soft water Substances 0.000 claims description 4
- 238000001179 sorption measurement Methods 0.000 claims description 4
- 239000007858 starting material Substances 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 239000008399 tap water Substances 0.000 claims description 4
- 235000020679 tap water Nutrition 0.000 claims description 4
- 241000589291 Acinetobacter Species 0.000 claims description 3
- 239000002131 composite material Substances 0.000 claims description 3
- 241000590020 Achromobacter Species 0.000 claims description 2
- 241000186063 Arthrobacter Species 0.000 claims description 2
- 241000186146 Brevibacterium Species 0.000 claims description 2
- 241000186216 Corynebacterium Species 0.000 claims description 2
- 241000589565 Flavobacterium Species 0.000 claims description 2
- 241000192041 Micrococcus Species 0.000 claims description 2
- 241000187654 Nocardia Species 0.000 claims description 2
- 241000589516 Pseudomonas Species 0.000 claims description 2
- FRYDSOYOHWGSMD-UHFFFAOYSA-N [C].O Chemical compound [C].O FRYDSOYOHWGSMD-UHFFFAOYSA-N 0.000 claims description 2
- 238000000926 separation method Methods 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract description 4
- 230000000968 intestinal effect Effects 0.000 abstract description 2
- 230000000593 degrading effect Effects 0.000 abstract 3
- 239000013543 active substance Substances 0.000 abstract 1
- 150000001450 anions Chemical class 0.000 abstract 1
- 239000008280 blood Substances 0.000 abstract 1
- 210000004369 blood Anatomy 0.000 abstract 1
- 230000008020 evaporation Effects 0.000 abstract 1
- 239000002244 precipitate Substances 0.000 abstract 1
- 238000010257 thawing Methods 0.000 abstract 1
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 229910052717 sulfur Inorganic materials 0.000 description 5
- 230000000813 microbial effect Effects 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 230000036541 health Effects 0.000 description 3
- 231100000252 nontoxic Toxicity 0.000 description 3
- 230000003000 nontoxic effect Effects 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- VEVZSMAEJFVWIL-UHFFFAOYSA-O cyanidin cation Chemical compound [O+]=1C2=CC(O)=CC(O)=C2C=C(O)C=1C1=CC=C(O)C(O)=C1 VEVZSMAEJFVWIL-UHFFFAOYSA-O 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000002101 lytic effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 241000588624 Acinetobacter calcoaceticus Species 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
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000035 biogenic effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000036983 biotransformation Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 235000007336 cyanidin Nutrition 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000037323 metabolic rate Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/582—Recycling of unreacted starting or intermediate materials
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a Microorganisms or/and biological enzyme extracting method for plant anthocyanin, belonging to the technical field of biological engineering. The method comprises the following steps of: classifying plants containing anthocyanin for smashing, adsorbing water, freezing the plants, heating the plants for thawing, pressing the plants, filtering for making juice, cracking and degrading the anthocyanin by the microorganisms or/and the biological enzyme cracking anthocyanin glycosidic bond, utilizing an anion active agent for precipitation reaction, degrading the precipitate by the microorganisms, and carrying out precipitation separation and evaporation concentration. Cation anthocyanin with the real content of more than 95%, namely color primitive but not the anthocyanin can be obtained by the technology of the invention. The cation anthocyanin made by the invention does not need human intestinal microorganisms for degrading, and is directly adsorbed and utilized by a human body or directly goes into human blood. The medical care value of the plant anthocyanin is much higher than that of the anthocyanin.
Description
Affiliated technical field:
The present invention relates to the extracting method of a kind of plant positively charged ion anthocyanidin of technical field of bioengineering, especially the microorganism of plant anthocyanidin is or/and biological enzyme extracting method.
Background technology:
Protect along with the raising of people's living standard with to nontoxic the helping in natural plant source that people are healthy, the pursuit of the natural organic substance of preventing and curing diseases; simultaneously also because synthetic anthocyanidin has bigger toxic side effect to human body; banned use of by the World Health Organization, so the natural phant positively charged ion anthocyanidin of high effect nontoxic and extracting method thereof have become 21 century emerging forward position research topic.
But existing separation and extraction technology just obtains containing the anthocyanogen of anthocyanidin, and content is too low, and impurity is too many, so it is used in the painted of food dyeing and makeup mostly, and value medical health care is low.Existing ultrasonic wave, microbiological deterioration are destroyed plant tissue cell's wall, extraction the separations purification technique of etc.ing extract the anthocyanogen that obtains and real to human body health care is arranged, the positively charged ion anthocyanidin that acts on of preventing and curing diseases is essentially different.Anthocyanogen is that positively charged ion anthocyanidin is that the pattern primitive combines the glucosides that forms with various monose by glycosidic link in the plant materials, this is a kind of neutral substance, biological activity is low, have only in human body that to form positively charged ion anthocyanidin by human enteric bacteria degraded desugar be the utilization that just can be absorbed by the body of pattern primitive, eliminate harmful free radicals in the human body, prevent and cure diseases, but this neutral anthocyanogen is degraded in human body, metabolic rate is very low, bioavailability is low, thereby human body is also very low to its specific absorption.
The individual bio-transformation function difference of adding the people is very big, and particularly the elderly and patient's enteron aisle is lower to the degraded and absorbed degree of anthocyanogen.Thereby the separation and Extraction of degraded of the outer industrialization plant anthocyanogen of human body and the positively charged ion anthocyanidin particularly important that just seems.This for fundamentally protect people healthy, preventing and curing diseases is of great immediate significance.
Summary of the invention:
For overcoming the defective of at present existing anthocyanogen separation and extraction technology effectively; really strong protection people healthy, prevent and cure diseases, the invention provides a kind of simple and reliable, technology is workable, production cost is low, the microorganism of effective, plant anthocyanidin that positively charged ion cyanidin extract purity is high is or/and biological enzyme extracting method.
The technical solution adopted for the present invention to solve the technical problems is:
1, extracting method of the present invention may further comprise the steps:
One, pre-treatment of raw material: the fresh starting material that contain plant anthocyanidin, as Pericarpium Vitis viniferae, Radix Dauci Sativae, indigo fruit, the soft water of classifying → pulverizes → eat soaks suction → freezing destruction plant tissue cell wall → heating and thaws → squeeze → filter and make standby (the 1) → residue of juice and suitably add water → soaking and stirring → squeezing → filtration and make juice standby (2).
Two, the above-mentioned squeeze and filter aqueous solution (1) and (2) are lumped together, and its water temperature is adjusted to following degraded microorganism or/and the suitableeest scope of biological enzyme.
Three, with above-mentioned product with the microorganism of cleavable anthocyanogen glycosidic link, degradable carbon hydrate or/and the biological enzyme biodegrade, first stirring at low speed, 100-600 rev/min, after leave standstill, make the anthocyanogen that contains anthocyanidin be degraded to positively charged ion anthocyanidin.The milk-acid bacteria of concrete available human enteric bacteria and bifidus bacillus etc. or black-koji mould are added one or more the composite synergys in beta-glucosidase, amygdalase, conversion carbohydrase, maltoside enzyme, beta-glycosidase, the pectin lyase.
Cracking and degradation time 10-24 hour, 25 ℃-40 ℃ of temperature, PH2.5-6.9.
More than various enzyme available starches substrate adsorption precipitation recyclings.
Need to use microorganism earlier before this step degraded or/and bio-enzyme degradation protein.
Four, above-mentioned product is added again the anion surfactant excessive a little, as ammonium lauryl sulfate (C with respect to positively charged ion anthocyanidin
12H
25OSO
3NH
4), stir with middling speed, 600-1400 rev/min, make it fully to leave standstill after the reaction, make till the most combined precipitation of positively charged ion anthocyanidin.This step reaction preferably is aided with the lower temperature more than 0 ℃, and this helps the quick and complete precipitation of reactant.
Five, take out above-mentioned aquo compound and carry out microbiological deterioration, can add a small amount of aseptic tap water, carry out in formerly leaving standstill after the stirring at low speed, about 100 rev/mins of stirring velocitys, the anion surfactant that is added is degraded by microorganisms, the cation activity of recovery anthocyanidin and water-soluble.
The combination biodegrade of one or more in concrete available Pseudomonas, achromobacter, genus arthrobacter, micrococcus sp, Nocardia, acinetobacter, brevibacterium sp, Corynebacterium, these microorganisms of Flavobacterium.
Degradation time 10-24 hour, 20 ℃-40 ℃ of degradation temperatures, PH3-8.
Six, with the sterilization of above-mentioned degraded product heat drying, 70 ℃-100 ℃ of drying temperatures get positively charged ion anthocyanidin product.
When seven, producing the pharmaceutical positively charged ion anthocyanidin of high purity product, can the residual body precipitation of inactivating proteins be separated out with aforesaid method gained anthocyanidin product with distilled water or edible ethanol dissolving.
Eight, to make highly purified positively charged ion anthocyanidin dry product, also need the further evaporating, concentrating and drying of above-mentioned anthocyanidin solution.
2, described extracting method is preferably:
It is made up of the following step:
One, pre-treatment of raw material: the fresh starting material that contain plant anthocyanidin, as Pericarpium Vitis viniferae, Radix Dauci Sativae, indigo fruit, the soft water of classifying → pulverizes → eat soak absorb water → freezing destruction plant tissue cell wall → heating thaws → squeezes → filter that to make juice standby
(1) → residue suitably adds water → soaking and stirring → squeezing → filtration system juice standby (2).
Two, the above-mentioned squeeze and filter aqueous solution (1) and (2) are lumped together, and its water temperature is adjusted to following degraded microorganism or/and the suitableeest scope of biological enzyme.
Three, add beta-glucosidase, amygdalase, conversion carbohydrase, maltoside enzyme, beta-glycosidase, the composite synergy of pectin lyase, biodegrade with the milk-acid bacteria of human enteric bacteria and bifidus bacillus etc.The elder generation stirring at low speed, 100-600 rev/min, after leave standstill, make the anthocyanogen that contains anthocyanidin be degraded to positively charged ion anthocyanidin.
Cracking and degradation time 10-24 hour, 25 ℃-40 ℃ of temperature, PH2.5-6.9.
More than various enzyme available starches substrate adsorption precipitation recyclings.
Need to use microorganism earlier before this step degraded or/and bio-enzyme degradation protein.
Four, above-mentioned product is added again the ammonium lauryl sulfate (C excessive a little with respect to positively charged ion anthocyanidin
12H
25OSO
3NH
4), stir with middling speed, 600-1400 rev/min, make it fully to leave standstill after the reaction, make till the most combined precipitation of positively charged ion anthocyanidin.This step reaction preferably is aided with the lower temperature more than 0 ℃, and this helps the quick and complete precipitation of reactant.
Five, take out above-mentioned aquo compound acinetobacter calcoaceticus microorganism belonging to genus biodegrade.
Can add a small amount of aseptic tap water, carry out in formerly leaving standstill after the stirring at low speed, about 100 rev/mins of stirring velocitys are degraded by microorganisms the anion surfactant that is added, the cation activity of recovery anthocyanidin and water-soluble.
Degradation time 10-24 hour, 20 ℃-40 ℃ of degradation temperatures, PH3-8.
Six, with the sterilization of above-mentioned degraded product heat drying, 70 ℃-100 ℃ of drying temperatures get positively charged ion anthocyanidin product.
When seven, producing the pharmaceutical positively charged ion anthocyanidin of high purity product, can the residual body precipitation of inactivating proteins be separated out with aforesaid method gained anthocyanidin product with distilled water or edible ethanol dissolving.
Eight, to make highly purified positively charged ion anthocyanidin dry product, also need the further evaporating, concentrating and drying of above-mentioned anthocyanidin solution.
The invention has the beneficial effects as follows:
1, the present invention help cation anthocyanidin plant resources effectively and reasonably and make full use of, reduce the waste and the waste discharge of plant resources, avoid environmental pollution.
2, the present invention helps extracting the anthocyanogen that obtains highly purified positively charged ion anthocyanidin product but not contain the low-purity of anthocyanidin.Realize the medical value of plant positively charged ion anthocyanidin, prevent and cure diseases, the medical first aid effect is remarkable.
3, technical solution of the present invention is simple, easy handling, and production cost is low, is easy to industrialization.
4, degraded product positively charged ion anthocyanidin of the present invention is promptly innovated essential features with anion surfactant combination reaction settling step and with the microorganism of anthocyanogen or/and the orderly combination of the microbiological deterioration step of biological enzyme cracking and degraded, positively charged ion anthocyanidin and anionic surfactant compound is a core technology of the present invention.
5, the present invention needs to carry out in no harmful microbe environment.
6, the selection of anion surfactant of the present invention requires:
(1) the nontoxic or low toxicity to microorganism; (2) molecular weight is less, and carbochain is lacked (C
10-C
22) contain the straight chain oleophilic group; (3) biological degradability is good, and degraded relatively rapidly, thoroughly; (4) these can be converted into the element of gaseous substance and water or keep the necessary element of microbial life activity only to contain C, H, O, S, N, P.
7, the temperature and the PH of the substrate of waiting to degrade to be regulated and control to the suitableeest scope of various microbial life active in microbial lytic of the present invention and the degradation process, and keep microorganism before cracking and degraded the growth and breeding stage and cracking and degradation process in have comprehensively, abundance and eurythmy keep its biogenic C, H, O, S, N, P etc. reach other each micro-element nutrition sources.
As with additional C sources such as glucose and starch that boils or pectin, also guide the glycosidic link of microbial lytic anthocyanogen with this.Replenish the N source with urea.With (NH
4)
3PO
4Replenish N and P.With (NH
4)
2SO
4Replenish S and N.With K
2SO
4Replenish S and K, with Ca (HCO
3)
2, Mg (HCO
3)
2Replenish Ca and Mg.With FeSO
4, Fe
2(SO
4)
3Replenish S and Fe.
8, extracting method of the present invention also is that other need the vegetalitas glycoside through human intestinal degraded, metabolism and absorption, carries out outer cracking of human body and degraded, the basic skills of its active aglycon of separation and Extraction or important references.
Embodiment:
Below just embodiment the invention will be further described:
Embodiment:
Carry out as follows during extraction:
One, pre-treatment of raw material: the fresh starting material that contain plant anthocyanidin, as Pericarpium Vitis viniferae, Radix Dauci Sativae, indigo fruit, the soft water of classifying → pulverizes → eat soaks suction → freezing destruction plant tissue cell wall → heating and thaws → squeeze → filter and make standby (the 1) → residue of juice and suitably add water → soaking and stirring → squeezing → filtration and make juice standby (2).
Two, the above-mentioned squeeze and filter aqueous solution (1) and (2) are lumped together, and its water temperature is adjusted to following degraded microorganism or/and the suitableeest scope of biological enzyme.
Three, above-mentioned product being added beta-glucosidase, amygdalase, conversion carbohydrase, maltoside enzyme, beta-glycosidase, pectin lyase with the milk-acid bacteria of human enteric bacteria and bifidus bacillus etc. is used, also above-mentioned human enteric bacteria and above-mentioned various enzymes can be separated use, and the adjusting pH value, biodegrade.The elder generation stirring at low speed, 100-600 rev/min, after leave standstill, make the anthocyanogen that contains anthocyanidin be degraded to positively charged ion anthocyanidin.
Cracking and degradation time 10-24 hour, 25 ℃-40 ℃ of temperature, PH2.5-6.9.
More than various enzyme available starches substrate adsorption precipitation recyclings.
Need to use microorganism earlier before this step degraded or/and bio-enzyme degradation protein.
Four, above-mentioned product is added again the ammonium lauryl sulfate (C excessive a little with respect to positively charged ion anthocyanidin
12H
25OSO
3NH
4), stir with middling speed, 600-1400 rev/min, make it fully to leave standstill after the reaction, make till the most combined precipitation of positively charged ion anthocyanidin.This step reaction preferably is aided with the lower temperature more than 0 ℃, and this helps the quick and complete precipitation of reactant.
Five, take out above-mentioned aquo compound these microorganism biodegrades of acinetobacter.
Can add a small amount of aseptic tap water, carry out in formerly leaving standstill after the stirring at low speed, about 100 rev/mins of stirring velocitys are degraded by microorganisms the anion surfactant that is added, the cation activity of recovery anthocyanidin and water-soluble.
Degradation time 10-24 hour, 20 ℃-40 ℃ of degradation temperatures, PH3-8.
Six, with the sterilization of above-mentioned degraded product heat drying, 70 ℃-100 ℃ of drying temperatures get positively charged ion anthocyanidin product.
When seven, producing the pharmaceutical positively charged ion anthocyanidin of high purity product, can the residual body precipitation of inactivating proteins be separated out with aforesaid method gained anthocyanidin product with distilled water or edible ethanol dissolving.
Eight, to make highly purified positively charged ion anthocyanidin dry product, also need the further evaporating, concentrating and drying of above-mentioned anthocyanidin solution.
Claims (1)
1. the microorganism of plant anthocyanidin is or/and biological enzyme extracting method, it is characterized in that it is made up of the following step:
One, pre-treatment of raw material: the fresh starting material that contain plant anthocyanidin, as Pericarpium Vitis viniferae, Radix Dauci Sativae, indigo fruit, the soft water of classifying → pulverizes → eat soaks suction → freezing destruction plant tissue cell wall → heating and thaws → squeeze → filter and make standby (the 1) → residue of juice and suitably add water → soaking and stirring → squeezing → filtration and make juice standby (2).
Two, the above-mentioned squeeze and filter aqueous solution (1) and (2) are lumped together, and its water temperature is adjusted to following degraded microorganism or/and the suitableeest scope of biological enzyme.
Three, with above-mentioned product with the microorganism of cleavable anthocyanogen glycosidic link, degradable carbon hydrate or/and the biological enzyme biodegrade, first stirring at low speed, 100-600 rev/min, after leave standstill, make the anthocyanogen that contains anthocyanidin be degraded to positively charged ion anthocyanidin.
The milk-acid bacteria of concrete available human enteric bacteria and bifidus bacillus etc. or black-koji mould are added one or more the composite synergys in beta-glucosidase, amygdalase, conversion carbohydrase, maltoside enzyme, beta-glycosidase, the pectin lyase.
Cracking and degradation time 10-24 hour, 25 ℃-40 ℃ of temperature, PH2.5-6.9.
More than various enzyme available starches substrate adsorption precipitation recyclings.
Need to use microorganism earlier before this step degraded or/and bio-enzyme degradation protein.
Four, above-mentioned product is added again the anion surfactant excessive a little, as ammonium lauryl sulfate (C with respect to positively charged ion anthocyanidin
12H
25OSO
3NH
4), stir with middling speed, 600-1400 rev/min, make it fully to leave standstill after the reaction, make till the most combined precipitation of positively charged ion anthocyanidin.This step reaction preferably is aided with the lower temperature more than 0 ℃, and this helps the quick and complete precipitation of reactant.
Five, take out above-mentioned aquo compound and carry out microbiological deterioration, can add a small amount of aseptic tap water, carry out in formerly leaving standstill after the stirring at low speed, about 100 rev/mins of stirring velocitys, the anion surfactant that is added is degraded by microorganisms, the cation activity of recovery anthocyanidin and water-soluble.
The combination biodegrade of one or more in concrete available Pseudomonas, achromobacter, genus arthrobacter, micrococcus sp, Nocardia, acinetobacter, brevibacterium sp, Corynebacterium, these microorganisms of Flavobacterium.
Degradation time 10-24 hour, 20 ℃-40 ℃ of degradation temperatures, PH3-8.
Six, with the sterilization of above-mentioned degraded product heat drying, 70 ℃-100 ℃ of drying temperatures get positively charged ion anthocyanidin product.
When seven, producing the pharmaceutical positively charged ion anthocyanidin of high purity product, can the residual body precipitation of inactivating proteins be separated out with aforesaid method gained anthocyanidin product with distilled water or edible ethanol dissolving.
Eight, to make highly purified positively charged ion anthocyanidin dry product, also need the further evaporating, concentrating and drying of above-mentioned anthocyanidin solution.
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