CN114214374A - Method for preparing soybean isoflavone aglycone by fermentation method - Google Patents
Method for preparing soybean isoflavone aglycone by fermentation method Download PDFInfo
- Publication number
- CN114214374A CN114214374A CN202111596962.4A CN202111596962A CN114214374A CN 114214374 A CN114214374 A CN 114214374A CN 202111596962 A CN202111596962 A CN 202111596962A CN 114214374 A CN114214374 A CN 114214374A
- Authority
- CN
- China
- Prior art keywords
- extract
- soybean isoflavone
- isoflavone aglycone
- aglycone
- complex enzyme
- 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
- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 106
- 244000068988 Glycine max Species 0.000 title claims abstract description 105
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 title claims abstract description 84
- 235000008696 isoflavones Nutrition 0.000 title claims abstract description 84
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 title claims abstract description 82
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 title claims abstract description 53
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 title claims abstract description 53
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 31
- 238000000855 fermentation Methods 0.000 title claims abstract description 10
- 230000004151 fermentation Effects 0.000 title claims abstract description 10
- 239000000284 extract Substances 0.000 claims abstract description 85
- 102000004190 Enzymes Human genes 0.000 claims abstract description 28
- 108090000790 Enzymes Proteins 0.000 claims abstract description 28
- 238000006243 chemical reaction Methods 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000012065 filter cake Substances 0.000 claims abstract description 14
- 238000001035 drying Methods 0.000 claims abstract description 9
- 239000002253 acid Substances 0.000 claims abstract description 8
- 238000007865 diluting Methods 0.000 claims abstract description 8
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 239000000463 material Substances 0.000 claims abstract description 7
- 239000003085 diluting agent Substances 0.000 claims abstract description 5
- 230000000415 inactivating effect Effects 0.000 claims abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 64
- 238000000605 extraction Methods 0.000 claims description 46
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 21
- 108010056771 Glucosidases Proteins 0.000 claims description 17
- 102000004366 Glucosidases Human genes 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 16
- 108010059892 Cellulase Proteins 0.000 claims description 13
- 101710112457 Exoglucanase Proteins 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000008213 purified water Substances 0.000 claims description 8
- 210000001161 mammalian embryo Anatomy 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims description 2
- 239000000243 solution Substances 0.000 description 16
- 239000000047 product Substances 0.000 description 11
- ZQSIJRDFPHDXIC-UHFFFAOYSA-N daidzein Chemical compound C1=CC(O)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZQSIJRDFPHDXIC-UHFFFAOYSA-N 0.000 description 10
- 210000002257 embryonic structure Anatomy 0.000 description 9
- 230000009261 transgenic effect Effects 0.000 description 9
- ZWSNUPOSLDAWJS-QNDFHXLGSA-N 6,7-dihydroxy-3-[4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]chromen-4-one Chemical compound OC[C@H]1O[C@@H](Oc2ccc(cc2)-c2coc3cc(O)c(O)cc3c2=O)[C@H](O)[C@@H](O)[C@@H]1O ZWSNUPOSLDAWJS-QNDFHXLGSA-N 0.000 description 7
- 238000006460 hydrolysis reaction Methods 0.000 description 7
- 235000003687 soy isoflavones Nutrition 0.000 description 6
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 description 5
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000005070 sampling Methods 0.000 description 5
- 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 5
- 238000005903 acid hydrolysis reaction Methods 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 235000007240 daidzein Nutrition 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- OZBAVEKZGSOMOJ-MIUGBVLSSA-N glycitin Chemical compound COC1=CC(C(C(C=2C=CC(O)=CC=2)=CO2)=O)=C2C=C1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O OZBAVEKZGSOMOJ-MIUGBVLSSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- ZCOLJUOHXJRHDI-FZHKGVQDSA-N Genistein 7-O-glucoside Natural products O([C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O1)c1cc(O)c2C(=O)C(c3ccc(O)cc3)=COc2c1 ZCOLJUOHXJRHDI-FZHKGVQDSA-N 0.000 description 3
- CJPNHKPXZYYCME-UHFFFAOYSA-N Genistin Natural products OCC1OC(Oc2ccc(O)c3OC(=CC(=O)c23)c4ccc(O)cc4)C(O)C(O)C1O CJPNHKPXZYYCME-UHFFFAOYSA-N 0.000 description 3
- YCUNGEJJOMKCGZ-UHFFFAOYSA-N Pallidiflorin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC(O)=C2C1=O YCUNGEJJOMKCGZ-UHFFFAOYSA-N 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 229930182478 glucoside Natural products 0.000 description 3
- 150000008131 glucosides Chemical group 0.000 description 3
- 229930182470 glycoside Natural products 0.000 description 3
- 150000002338 glycosides Chemical class 0.000 description 3
- SHHLMGCHMMCOOS-UHFFFAOYSA-N 4h-chromen-3-one Chemical compound C1=CC=C2CC(=O)COC2=C1 SHHLMGCHMMCOOS-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- GMTUGPYJRUMVTC-UHFFFAOYSA-N Daidzin Natural products OC(COc1ccc2C(=O)C(=COc2c1)c3ccc(O)cc3)C(O)C(O)C(O)C=O GMTUGPYJRUMVTC-UHFFFAOYSA-N 0.000 description 2
- KYQZWONCHDNPDP-UHFFFAOYSA-N Daidzoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-UHFFFAOYSA-N 0.000 description 2
- XJTZHGNBKZYODI-UHFFFAOYSA-N Glycitin Natural products OCC1OC(Oc2ccc3OC=C(C(=O)c3c2CO)c4ccc(O)cc4)C(O)C(O)C1O XJTZHGNBKZYODI-UHFFFAOYSA-N 0.000 description 2
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- RXRFEELZASHOLV-JAJWTYFOSA-N [(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] acetate Chemical group CC(=O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RXRFEELZASHOLV-JAJWTYFOSA-N 0.000 description 2
- 108010047754 beta-Glucosidase Proteins 0.000 description 2
- 102000006995 beta-Glucosidase Human genes 0.000 description 2
- FYGDTMLNYKFZSV-ZWSAEMDYSA-N cellotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](OC(O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-ZWSAEMDYSA-N 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- KYQZWONCHDNPDP-QNDFHXLGSA-N daidzein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-QNDFHXLGSA-N 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000006539 genistein Nutrition 0.000 description 2
- TZBJGXHYKVUXJN-UHFFFAOYSA-N genistein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O TZBJGXHYKVUXJN-UHFFFAOYSA-N 0.000 description 2
- 229940045109 genistein Drugs 0.000 description 2
- 150000002333 glycines Chemical class 0.000 description 2
- 235000008466 glycitein Nutrition 0.000 description 2
- NNUVCMKMNCKPKN-UHFFFAOYSA-N glycitein Natural products COc1c(O)ccc2OC=C(C(=O)c12)c3ccc(O)cc3 NNUVCMKMNCKPKN-UHFFFAOYSA-N 0.000 description 2
- DXYUAIFZCFRPTH-UHFFFAOYSA-N glycitein Chemical compound C1=C(O)C(OC)=CC(C2=O)=C1OC=C2C1=CC=C(O)C=C1 DXYUAIFZCFRPTH-UHFFFAOYSA-N 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- FRAUJUKWSKMNJY-UHFFFAOYSA-N 5-hydroxy-3-(4-hydroxyphenyl)-7-(6-malonyl-beta-D-glucopyranosyloxy)-4H-1-benzopyran-4-one Natural products OC1C(O)C(O)C(COC(=O)CC(O)=O)OC1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 FRAUJUKWSKMNJY-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 229940124443 chemopreventive agent Drugs 0.000 description 1
- 239000012627 chemopreventive agent Substances 0.000 description 1
- 238000003795 desorption Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 230000001076 estrogenic effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- RTRZOHKLISMNRD-UHFFFAOYSA-N isoflavanone Chemical compound C1OC2=CC=CC=C2C(=O)C1C1=CC=CC=C1 RTRZOHKLISMNRD-UHFFFAOYSA-N 0.000 description 1
- 150000002515 isoflavone derivatives Chemical class 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- MTXMHWSVSZKYBT-UHFFFAOYSA-N malonyl daidzin Natural products OC1C(O)C(O)C(COC(=O)CC(O)=O)OC1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 MTXMHWSVSZKYBT-UHFFFAOYSA-N 0.000 description 1
- -1 malonyl glucoside Chemical class 0.000 description 1
- MTXMHWSVSZKYBT-ASDZUOGYSA-N malonyldaidzin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](COC(=O)CC(O)=O)O[C@H]1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 MTXMHWSVSZKYBT-ASDZUOGYSA-N 0.000 description 1
- FRAUJUKWSKMNJY-RSEYPYQYSA-N malonylgenistin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](COC(=O)CC(O)=O)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 FRAUJUKWSKMNJY-RSEYPYQYSA-N 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 235000017807 phytochemicals Nutrition 0.000 description 1
- 239000003075 phytoestrogen Substances 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/34—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 3 only
- C07D311/36—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 3 only not hydrogenated in the hetero ring, e.g. isoflavones
Abstract
The invention discloses a method for preparing soybean isoflavone aglycone by a fermentation method, which comprises the following steps: s21, taking soybean isoflavone, water and conversion complex enzyme; s22, diluting the soybean isoflavone with water, adjusting the pH of the diluent with acid, adding a conversion complex enzyme, and performing enzymolysis to obtain an extract; s23, inactivating the extract; s24, filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake; and S25, drying the filter cake, and crushing the dried material. The soybean isoflavone aglycone prepared by the method has high content and conversion rate.
Description
Technical Field
The invention relates to a preparation method of soybean isoflavone aglycone by a fermentation method and a preparation method of the soybean isoflavone aglycone.
Background
Soy isoflavones are flavonoids, a class of secondary metabolites formed during the growth of soybeans, and are bioactive substances. Since it is extracted from plants and has a structure similar to that of estrogen, soy isoflavones are also called phytoestrogens. The estrogenic action of soy isoflavones affects hormone secretion, metabolic biological activity, protein synthesis, growth factor activity, and is a natural cancer chemopreventive agent.
Soybean Isoflavone (Soy Isoflavanone) is a phytochemical, belongs to plant flavonoid, is mainly derived from legume of leguminous plant, and has high content of soybean of 0.1% -0.5%. Mainly refers to a compound taking 3-benzopyrone as a mother nucleus, and the total of the naturally occurring soybean isoflavones in soybeans is 12, which can be divided into 3 types, namely glycitin (Daidzingroups), genistin (Genistingroups) and glycitin (glycitinroups). Each group exists in 4 forms such as free form, glucoside form, acetyl glucoside form, malonyl glucoside form, etc. The aglycone (Aglycon) in free form accounts for 2-3% of the total amount, and comprises Genistein (Genistein), Daidzein (Daidzein) and glycitein (glycitein). The conjugated Glycosides (Glycosides) account for 97-98% of the total amount, and are present mainly in the form of genistin and Daidzin (Daidzin) and malonyl genistin (6 '-O-ma-1 onylGenistin) and malonyl Daidzin (6' -O-malonyldaid-zin), accounting for about 95% of the total amount. The planting environment, the processing method, the genetic factors and the like have certain influence on the content and the components of the soybean isoflavone, and the influence is shown as the difference of the total amount of the isoflavone and the proportion of each component in different soybean varieties.
The main structural form of soybean isoflavone is a compound group taking 3-benzopyrone as a mother nucleus. The natural soybean isoflavone is mainly divided into free type aglycone and combined type glucoside. Wherein the bound glycoside is deglycosylated by hydrolysis with enzyme or dilute acid to form soy isoflavone aglycone. The free type soybean isoflavone aglycone includes genistin, daidzein and daidzein. The bound soybean isoflavone glycoside is glucoside type, acetyl glucoside type, or malonyl glucoside type of the above 3 aglycones.
At present, the production method of soybean isoflavone is as follows: A. extraction: extracting with ethanol for several times in a multifunctional extraction tank, mixing extractive solutions, and concentrating to obtain crude extract; B. diluting the crude extract with a large amount of water, adsorbing with resin column, desorbing with high concentration ethanol to obtain solution, mixing the concentrated solutions to obtain solution water solution; C. and crystallizing the aqueous solution of the desorption solution, and centrifuging to obtain the product. The process is simple in production, but the content of the obtained finished product is low, and a large amount of sewage is generated in the production process. Adversely affecting environmental protection.
The soybean isoflavone glycoside can be absorbed into human body to exert its function only after being converted into soybean isoflavone aglycone, and although the efficacy of soybean isoflavone on human body health is highly concerned, the soybean isoflavone aglycone has a small proportion in soybean isoflavone, so that the separation and purification difficulty is high, the yield is low and the cost is high.
At present, the method for converting soybean isoflavone glycoside into soybean isoflavone aglycone mainly comprises two methods, namely acid hydrolysis and enzyme hydrolysis. The acid hydrolysis method requires high temperature, has poor specificity in hydrolysis reaction, can destroy other groups on isoflavone besides hydrolysis glycosidic bond, causes unstable structure of isoflavone aglycone to cause activity disappearance, and pollutes the environment by acid hydrolysis, so the research on the acid hydrolysis method is more, but the application effect is not ideal. The enzyme hydrolysis method is to hydrolyze the beta-glycosidic bond of the soybean isoflavone glycoside by using beta-glucosidase to generate the soybean isoflavone aglycone, and the hydrolysis reaction has the advantage of beta-glycosidic bond specificity, but on one hand, the method also stays in the stage of researching the isoflavone glycoside hydrolysis by free enzyme, the enzyme stability is poor, and a microbial strain which needs to produce the beta-glucosidase is needed. On the other hand, the conversion rate and the purity of the soybean isoflavone aglycone product are lower. CN200910116404.6 discloses a method for producing soybean isoflavone aglycone by magnetic nano immobilized enzyme catalysis, which converts soybean isoflavone glycoside into soybean isoflavone aglycone by magnetic nano immobilized enzyme, but the conversion rate is low, and the purity of the soybean isoflavone aglycone product is low.
Disclosure of Invention
Based on the problems, the invention provides a method for preparing soybean isoflavone aglycone by a fermentation method, and the content and the conversion rate of the soybean isoflavone aglycone prepared by the method are high.
A method for preparing soybean isoflavone aglycone by fermentation comprises the following steps:
s21, taking soybean isoflavone, water and conversion complex enzyme;
s22, diluting the soybean isoflavone with water, adjusting the pH of the diluent with acid, adding a conversion complex enzyme, and performing enzymolysis to obtain an extract;
s23, inactivating the extract;
s24, filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake;
and S25, drying the filter cake, and crushing the dried material.
Further, the conversion complex enzyme is a mixture of endoglucanase, exoglucanase and glucosidase, wherein the endoglucanase: exoglucanase: the weight ratio of the glucosidase to the glucosidase is 0.01-99.5: 0.01-99.5: 0.01-99.5.
Further, in the S22, the pH value is 2-6.
Further, the enzymolysis temperature is 20-70 ℃, and the enzymolysis time is 24-72 h.
The invention also provides a preparation method of the soybean isoflavone aglycone.
A method for preparing soybean isoflavone aglycone comprises the following steps:
s1, preparing extract of the extract liquid:
s11, taking defatted embryo bud of semen glycines and ethanol, and feeding the defatted embryo bud of semen glycines and ethanol into continuous countercurrent extraction equipment for continuous countercurrent extraction to obtain extractive solution;
s12, concentrating the extracting solution and recovering ethanol to obtain an extract;
s13, extracting the extract by using ethyl acetate to obtain extract liquid;
s14, drying the extract to obtain soybean isoflavone;
s2, preparation of soybean isoflavone aglycone:
s21, taking the extract liquid extract prepared in the S13, purified water and conversion complex enzyme;
s22, diluting the extract with purified water, adjusting the pH of the diluent with acid, adding a conversion complex enzyme, and obtaining an extract;
s23, inactivating the extract;
s24, filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake;
and S25, drying the filter cake, and crushing the dried material.
Further, the conversion complex enzyme is a mixture of endoglucanase, exoglucanase and glucosidase, wherein the endoglucanase: exoglucanase: the weight ratio of the glucosidase to the glucosidase is 0.01-99.5: 0.01-99.5: 0.01-99.5.
Further, in the S22, the pH value is 2-6, the enzymolysis temperature is 20-70 ℃, and the enzymolysis time is 24-72 h.
Further, in the step S11, the extraction time of the continuous countercurrent extraction is 2-10 h.
Further, the continuous countercurrent extraction temperature is controlled at 40-70 ℃; in the S12, the concentration recovery temperature is 60-80 ℃.
Further, in the above S13, the extraction temperature was 55 to 60 ℃.
The invention principle and the beneficial effects are as follows:
in the extraction stage, the defatted germs of the soybeans are extracted in a continuous countercurrent mode to improve the extraction rate of the soybean isoflavone glycoside; in the conversion stage, the soybean isoflavone is converted into the soybean isoflavone aglycone by converting complex enzyme, wherein cellulose is degraded into amorphous cellulose by endoglucanase, the amorphous cellulose is hydrolyzed into cellooligosaccharide by exoglucanase, the cellooligosaccharide is hydrolyzed into monosaccharide by glucosidase, glycosyl on soybean isoflavone glycosyl is removed, the conversion rate of the soybean isoflavone aglycone reaches more than 99.5 percent, and the purity of the soybean isoflavone aglycone product is more than 90 percent.
Detailed Description
The present invention will be further explained below.
In this embodiment, the ethanol is 95% ethanol that is commercially available.
A method for producing a soy isoflavone aglycone product, comprising the steps of:
s1, preparing extract of the extract, which comprises the following steps:
s11, taking the defatted embryo bud of soybean and ethanol (ethanol is used as extractant), feeding the defatted embryo bud of soybean and ethanol into a continuous countercurrent extraction device for continuous countercurrent extraction for 2-10h, and controlling the temperature of the continuous countercurrent extraction device at 40-70 ℃ to obtain extract.
S12, concentrating the extract at 60-80 deg.C, recovering ethanol to obtain extract, sampling, and detecting to obtain soybean isoflavone.
S13, extracting the extract by using ethyl acetate at the extraction temperature of 55-60 ℃ for 3 times, and combining and concentrating the extract to obtain the extract.
S2, the preparation of soybean isoflavone aglycone comprises the following steps:
and S21, taking the extract liquid extract prepared in the S13, purified water and a conversion complex enzyme (the conversion complex enzyme is a mixture of endoglucanase, exoglucanase and glucosidase, and the weight ratio of the endoglucanase to the exoglucanase to the glucosidase is 0.01-99.5: 0.01-99.5: 0.01-99.5).
S22, diluting the extract with purified water, adjusting pH of the diluted solution to 2-6 with acid, adding converting complex enzyme, and performing enzymolysis at 20-70 deg.C for 24-72 h.
S23, heating to 80-100 ℃ for 1-3h after enzymolysis to deactivate the conversion complex enzyme.
And S24, filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake.
S25, drying the filter cake, and crushing the dried material to obtain the soybean isoflavone aglycone product.
In S12, the content of soybean isoflavone in the extract is not less than 6%, and the extraction rate of soybean isoflavone is not less than 99.2%.
In S13, the extract has soybean isoflavone content of 80-90% and aglycon soybean isoflavone aglycon content of 2-3%.
In S25, the soybean isoflavone aglycone product has a soybean isoflavone aglycone content of 90-98.5% and a conversion rate of 99.5-99.8%.
EXAMPLE 1 preparation of extract
Taking 100g of defatted embryos of northeast non-transgenic soybeans and 600ml of ethanol (ethanol is used as an extracting agent), and feeding the defatted embryos of the northeast non-transgenic soybeans and the ethanol into continuous countercurrent extraction equipment for continuous countercurrent extraction, wherein the temperature of the continuous countercurrent extraction equipment is controlled at 50 ℃ to obtain an extracting solution.
The continuous countercurrent extraction time was 9 h.
② concentrating the extracting solution at 70 ℃ and recovering ethanol to obtain extract, sampling and detecting the extract, the content of the soybean isoflavone is 6.18 percent, and the extraction rate of the soybean isoflavone is 99.5 percent.
③ extracting the extract by 200ml ethyl acetate for 3 times at the extraction temperature of 55-60 ℃, and merging and concentrating the extract to obtain the extract. The extract liquid is sampled and detected, the content of the soybean isoflavone is 88.96 percent, and the content of the aglycone soybean isoflavone aglycone is 2.05 percent.
In this embodiment, ethanol recovered by concentration may be reused as the extract.
Example 2
And (3) centrifugally separating and drying the extract liquid extract prepared in the example 1 to obtain a soybean isoflavone product.
EXAMPLE 3 preparation of Soy isoflavone aglycone
Taking 100g of defatted embryos of northeast non-transgenic soybeans and 600ml of ethanol (ethanol is used as an extracting agent), and feeding the defatted embryos of the northeast non-transgenic soybeans and the ethanol into continuous countercurrent extraction equipment for continuous countercurrent extraction, wherein the temperature of the continuous countercurrent extraction equipment is controlled at 50 ℃ to obtain an extracting solution.
The continuous countercurrent extraction time was 10 h.
② concentrating the extracting solution at 70 ℃ and recovering ethanol to obtain extract, sampling and detecting the extract, the content of soybean isoflavone is 6.23 percent, and the extraction rate of soybean isoflavone is 99.6 percent.
③ extracting the extract by 200ml ethyl acetate for 3 times at the extraction temperature of 55-60 ℃, and merging and concentrating the extract to obtain the extract. The extract liquid is sampled and detected, the content of the soybean isoflavone is 88.86 percent, and the content of the aglycone soybean isoflavone aglycone is 2.01 percent.
And fourthly, taking the extract liquid extract prepared by the third step, purified water which is 5 times the weight of the extract liquid extract and conversion complex enzyme which is 8 per mill of the weight of the extract liquid extract (the conversion complex enzyme is a mixture of endoglucanase, exoglucanase and glucosidase, and the weight ratio of the endoglucanase to the exoglucanase to the glucosidase is 0.01-99.5: 0.01-99.5: 0.01-99.5).
Diluting the extract with purified water, adjusting pH of the diluted solution to 2-6 with acid, adding converting complex enzyme, and performing enzymolysis at 50 deg.C for 24 times.
Sixthly, after enzymolysis, the temperature is raised to 80-100 ℃ for 1-3h to ensure that the converted complex enzyme loses activity.
And filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake.
The filter cake is dried, the dried material is crushed to obtain the soybean isoflavone aglycone product, and the sampling detection of the soybean isoflavone aglycone product shows that the content of the soybean isoflavone aglycone is 96.48 percent and the conversion rate is 9.6 percent.
Comparative example 1
Taking 100g of defatted embryos of northeast non-transgenic soybeans and 600ml of ethanol (ethanol is used as an extracting agent), and feeding the defatted embryos of the northeast non-transgenic soybeans and the ethanol into an extraction tank, wherein the temperature of the extraction tank is controlled at 50 ℃, and the extraction time is 10 hours, so as to obtain an extracting solution.
② concentrating the extracting solution at 70 ℃ and recovering ethanol to obtain extract, sampling and detecting the extract, the content of the soybean isoflavone is 4.86 percent, and the extraction rate of the soybean isoflavone is 91.4 percent.
③ extracting the extract by 200ml ethyl acetate for 3 times at the extraction temperature of 55-60 ℃, and merging and concentrating the extract to obtain the extract. The extract liquid is sampled and detected, the content of the soybean isoflavone is 48.7 percent, and the content of the aglycon soybean isoflavone aglycone is 2.04 percent.
Comparative example 2
The method comprises the following steps of taking 100g of defatted embryos of northeast non-transgenic soybeans and 600ml of ethanol (ethanol is used as an extracting agent), fully wetting the defatted embryos of the northeast non-transgenic soybeans and the ethanol, and then putting the fully wetted defatted embryos of the northeast non-transgenic soybeans and the ethanol into an extraction kettle of a supercritical fluid extraction device for extraction, wherein the extraction pressure is 30MPa, the extraction temperature is 50 ℃, the carbon dioxide flow is 6L/h, and the extraction time is 10 hours, so that primary extraction liquid is obtained.
② concentrating the primary extract at 70 ℃ to recycle ethanol to obtain primary extract, detecting the primary extract sample, the content of soybean isoflavone is 4.78%, and the extraction rate of soybean isoflavone is 90.4%.
③ extracting the primary extract by 200g of ethyl acetate for 3 times at the extraction temperature of 55-60 ℃, and merging and concentrating the extract to obtain the extract. The extract liquid extract is sampled and detected, the content of soybean isoflavone is 47.5 percent, and the content of aglycon soybean isoflavone aglycone is 2.12 percent.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for preparing soybean isoflavone aglycone by fermentation comprises the following steps:
s21, taking soybean isoflavone, water and conversion complex enzyme;
s22, diluting the soybean isoflavone with water, adjusting the pH of the diluent with acid, adding a conversion complex enzyme, and performing enzymolysis to obtain an extract;
s23, inactivating the extract;
s24, filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake;
and S25, drying the filter cake, and crushing the dried material.
2. The method for preparing soybean isoflavone aglycone by fermentation according to claim 1, wherein the conversion complex enzyme is a mixture of endoglucanase, exoglucanase and glucosidase, and the endoglucanase: exoglucanase: the weight ratio of the glucosidase to the glucosidase is 0.01-99.5: 0.01-99.5: 0.01-99.5.
3. The method for preparing soybean isoflavone aglycone by fermentation according to claim 1, wherein pH is 2-6 in S22.
4. The method for preparing soybean isoflavone aglycone by fermentation according to claim 1, wherein the enzymolysis temperature is 20-70 ℃ and the enzymolysis time is 24-72 h.
5. A method for preparing soybean isoflavone aglycone comprises the following steps:
s1, preparing extract of the extract liquid:
s11, taking the defatted embryo bud of the soybean and ethanol, and feeding the defatted embryo bud of the soybean and the ethanol into continuous countercurrent extraction equipment for continuous countercurrent extraction to obtain an extracting solution;
s12, concentrating the extracting solution and recovering ethanol to obtain an extract;
s13, extracting the extract by using ethyl acetate to obtain extract liquid;
s14, drying the extract to obtain soybean isoflavone;
s2, preparation of soybean isoflavone aglycone:
s21, taking the extract liquid extract prepared in the S13, purified water and conversion complex enzyme;
s22, diluting the extract with purified water, adjusting the pH of the diluent with acid, adding a conversion complex enzyme, and obtaining an extract;
s23, inactivating the extract;
s24, filtering the inactivated extract by using a plate-and-frame filter to obtain a filter cake;
and S25, drying the filter cake, and crushing the dried material.
6. The process of claim 5, wherein the conversion complex enzyme is a mixture of endoglucanase, exoglucanase and glucosidase, and the ratio of endoglucanase: exoglucanase: the weight ratio of the glucosidase to the glucosidase is 0.01-99.5: 0.01-99.5: 0.01-99.5.
7. The method for preparing soybean isoflavone aglycone according to claim 5, wherein the pH of S22 is 2-6, the enzymolysis temperature is 20-70 deg.C, and the enzymolysis time is 24-72 h.
8. The method for preparing soybean isoflavone aglycone according to claim 5, wherein the extraction time of the continuous countercurrent extraction in S11 is 2-10 h.
9. The method for preparing soybean isoflavone aglycone according to claim 5, wherein the temperature of the continuous countercurrent extraction is controlled to 40-70 ℃; in the S12, the concentration recovery temperature is 60-80 ℃.
10. The method for preparing soybean isoflavone aglycone according to claim 5, wherein the extraction temperature of S13 is 55-60 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111596962.4A CN114214374A (en) | 2021-12-24 | 2021-12-24 | Method for preparing soybean isoflavone aglycone by fermentation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111596962.4A CN114214374A (en) | 2021-12-24 | 2021-12-24 | Method for preparing soybean isoflavone aglycone by fermentation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114214374A true CN114214374A (en) | 2022-03-22 |
Family
ID=80705537
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111596962.4A Pending CN114214374A (en) | 2021-12-24 | 2021-12-24 | Method for preparing soybean isoflavone aglycone by fermentation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114214374A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1566352A (en) * | 2003-06-12 | 2005-01-19 | 济南三株药业有限公司 | Process for preparing isoflavone aglycon |
| CN101532039A (en) * | 2009-03-25 | 2009-09-16 | 合肥工业大学 | Magnetic nano immobilized enzyme catalysis production method of soya isoflavone |
| CN101974577A (en) * | 2010-09-27 | 2011-02-16 | 徐州技源天然保健品有限公司 | Novel production process for extracting soy isoflavones aglycones |
| CN103536645A (en) * | 2013-10-17 | 2014-01-29 | 广东省天宝生物制药有限公司 | Method for preparing soy isoflavone aglycones and application of soy isoflavone aglycones to livestock breeding |
-
2021
- 2021-12-24 CN CN202111596962.4A patent/CN114214374A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1566352A (en) * | 2003-06-12 | 2005-01-19 | 济南三株药业有限公司 | Process for preparing isoflavone aglycon |
| CN101532039A (en) * | 2009-03-25 | 2009-09-16 | 合肥工业大学 | Magnetic nano immobilized enzyme catalysis production method of soya isoflavone |
| CN101974577A (en) * | 2010-09-27 | 2011-02-16 | 徐州技源天然保健品有限公司 | Novel production process for extracting soy isoflavones aglycones |
| CN103536645A (en) * | 2013-10-17 | 2014-01-29 | 广东省天宝生物制药有限公司 | Method for preparing soy isoflavone aglycones and application of soy isoflavone aglycones to livestock breeding |
Non-Patent Citations (4)
| Title |
|---|
| 李丹等: "纤维素酶水解大豆异黄酮糖苷混合物的抗氧化活性", 《食品研究与开发》 * |
| 王松等: "连续逆流法提取大豆异黄酮的研究", 《食品科技》 * |
| 钱丽丽等: "纤维素酶法水解大豆异黄酮糖苷研究", 《中国粮油学报》 * |
| 陈庆庆等: "纤维素酶对豆粕异黄酮提取的影响", 《化工学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1254131B8 (en) | Extraction of flavonoids | |
| CN109646992B (en) | Method for extracting cannabidiol concentrate from industrial cannabis sativa | |
| US7507423B2 (en) | Extraction of flavonoids | |
| EP1838862B1 (en) | Manufacturing method of kaempferol | |
| CN101967137A (en) | Method for extracting plant flavone compounds by enzymatic process | |
| CN101928273A (en) | Method for extracting and separating isoflavone from soybeans | |
| CN101974577A (en) | Novel production process for extracting soy isoflavones aglycones | |
| CN102002083A (en) | Method for extracting high-purity rutin with flash-extraction technology | |
| CN111187328B (en) | Method for preparing mogrol | |
| CN114214374A (en) | Method for preparing soybean isoflavone aglycone by fermentation method | |
| CN114315782A (en) | Preparation method of soybean isoflavone | |
| CN103356740A (en) | Preparation method of baicalein and scutellaria baicalensis flavone total-aglycone extractives | |
| CN100439364C (en) | Process for producing high purity matrine oxide | |
| CN107326059A (en) | A kind of preparation method and applications of the high activity aging ingredient of natural origin | |
| CN112300234A (en) | Method for extracting isoflavone, isoflavone extracted by method and application of isoflavone | |
| KR100379642B1 (en) | A process of manufacturing highly purified isoflavone aglycones by fermentation | |
| CN115043889B (en) | Method for extracting synephrine, hesperidin and naringin from seville orange flower | |
| CN101250177B (en) | Method for preparing quercetin from cotton petal | |
| CN113754626B (en) | Method for preparing fisetin by enzyme method | |
| CN107173803A (en) | The preparation method and application of bamboo-leaves flavones | |
| CN111575330B (en) | Method for hydrolyzing epimedium extract by plant-derived enzyme | |
| CN110903168B (en) | Method for subcritical extraction of solanesol in waste tobacco leaves | |
| CN113201034A (en) | Obtaining high-purity stevioside from primary crystallization mother liquor of stevioside through secondary crystallization and enriching rebaudioside C | |
| KR100346818B1 (en) | Process for producing aglycone isoflavones | |
| KR20150075647A (en) | Method for producing isoflavones aglycone from soybean embryo |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220322 |