CN111317694A - Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics - Google Patents
Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics Download PDFInfo
- Publication number
- CN111317694A CN111317694A CN202010143510.XA CN202010143510A CN111317694A CN 111317694 A CN111317694 A CN 111317694A CN 202010143510 A CN202010143510 A CN 202010143510A CN 111317694 A CN111317694 A CN 111317694A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- eucommia ulmoides
- eucommia
- powder
- enzymolysis
- 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.)
- Granted
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 199
- 230000004151 fermentation Effects 0.000 title claims abstract description 197
- 238000002360 preparation method Methods 0.000 title claims abstract description 35
- 239000002537 cosmetic Substances 0.000 title claims abstract description 17
- 241000208689 Eucommia ulmoides Species 0.000 title claims abstract 24
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 71
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 71
- 238000000034 method Methods 0.000 claims abstract description 62
- 239000000284 extract Substances 0.000 claims abstract description 54
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 53
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 53
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 53
- 241000589212 Acetobacter pasteurianus Species 0.000 claims abstract description 51
- 239000007788 liquid Substances 0.000 claims abstract description 48
- 239000000843 powder Substances 0.000 claims abstract description 48
- 239000000758 substrate Substances 0.000 claims abstract description 42
- 239000002131 composite material Substances 0.000 claims abstract description 36
- 108010059892 Cellulase Proteins 0.000 claims abstract description 27
- 108010059820 Polygalacturonase Proteins 0.000 claims abstract description 27
- 239000004365 Protease Substances 0.000 claims abstract description 27
- 229940106157 cellulase Drugs 0.000 claims abstract description 27
- 108010093305 exopolygalacturonase Proteins 0.000 claims abstract description 27
- 108091005804 Peptidases Proteins 0.000 claims abstract description 26
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000001301 oxygen Substances 0.000 claims abstract description 26
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 26
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 25
- 238000000746 purification Methods 0.000 claims abstract description 18
- 238000001914 filtration Methods 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 241000894006 Bacteria Species 0.000 claims description 39
- 102000004190 Enzymes Human genes 0.000 claims description 37
- 108090000790 Enzymes Proteins 0.000 claims description 37
- 229940088598 enzyme Drugs 0.000 claims description 35
- 238000012258 culturing Methods 0.000 claims description 30
- 230000008569 process Effects 0.000 claims description 28
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 26
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 22
- 230000000694 effects Effects 0.000 claims description 22
- 229910052799 carbon Inorganic materials 0.000 claims description 20
- 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 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 239000011780 sodium chloride Substances 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 239000003651 drinking water Substances 0.000 claims description 9
- 235000020188 drinking water Nutrition 0.000 claims description 9
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 238000011081 inoculation Methods 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 5
- 230000003301 hydrolyzing effect Effects 0.000 claims description 5
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 4
- 241000589220 Acetobacter Species 0.000 claims description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 239000012137 tryptone Substances 0.000 claims description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose 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](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 claims description 2
- 235000015278 beef Nutrition 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 2
- 229930182830 galactose Natural products 0.000 claims description 2
- 244000237330 gutta percha tree Species 0.000 description 141
- 239000000243 solution Substances 0.000 description 51
- 241000208688 Eucommia Species 0.000 description 50
- 230000000052 comparative effect Effects 0.000 description 26
- 235000019419 proteases Nutrition 0.000 description 22
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 18
- 238000002156 mixing Methods 0.000 description 18
- 230000001954 sterilising effect Effects 0.000 description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 14
- 239000002994 raw material Substances 0.000 description 14
- 230000002195 synergetic effect Effects 0.000 description 14
- 241000196324 Embryophyta Species 0.000 description 13
- 244000269722 Thea sinensis Species 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 13
- 235000013616 tea Nutrition 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 241001052560 Thallis Species 0.000 description 10
- 238000000605 extraction Methods 0.000 description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 239000000126 substance Substances 0.000 description 10
- 235000000346 sugar Nutrition 0.000 description 10
- 239000004480 active ingredient Substances 0.000 description 9
- 230000003321 amplification Effects 0.000 description 9
- 238000001816 cooling Methods 0.000 description 9
- 230000036541 health Effects 0.000 description 9
- 238000003199 nucleic acid amplification method Methods 0.000 description 9
- 238000009423 ventilation Methods 0.000 description 9
- 241000282414 Homo sapiens Species 0.000 description 8
- 230000036983 biotransformation Effects 0.000 description 8
- 238000009835 boiling Methods 0.000 description 8
- 238000004140 cleaning Methods 0.000 description 8
- 238000001035 drying Methods 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 210000003491 skin Anatomy 0.000 description 7
- 241000894007 species Species 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 239000006041 probiotic Substances 0.000 description 6
- 235000018291 probiotics Nutrition 0.000 description 6
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 5
- 241000186660 Lactobacillus Species 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 230000002255 enzymatic effect Effects 0.000 description 5
- 229930003935 flavonoid Natural products 0.000 description 5
- 235000017173 flavonoids Nutrition 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 229940039696 lactobacillus Drugs 0.000 description 5
- 229920001277 pectin Polymers 0.000 description 5
- 244000063299 Bacillus subtilis Species 0.000 description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 description 4
- 102000003777 Interleukin-1 beta Human genes 0.000 description 4
- 108090000193 Interleukin-1 beta Proteins 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000007547 defect Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 235000019985 fermented beverage Nutrition 0.000 description 4
- -1 flavonoid compounds Chemical class 0.000 description 4
- 150000002215 flavonoids Chemical class 0.000 description 4
- 150000004676 glycans Chemical class 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000036039 immunity Effects 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 239000001814 pectin Substances 0.000 description 4
- 235000010987 pectin Nutrition 0.000 description 4
- 229920001282 polysaccharide Polymers 0.000 description 4
- 239000005017 polysaccharide Substances 0.000 description 4
- 230000028327 secretion Effects 0.000 description 4
- ZJSJQWDXAYNLNS-UHFFFAOYSA-N (+) pinoresinol-4,4'-di-O-beta-D-glucopyranoside Natural products COC1=CC(C2C3C(C(OC3)C=3C=C(OC)C(OC4C(C(O)C(O)C(CO)O4)O)=CC=3)CO2)=CC=C1OC1OC(CO)C(O)C(O)C1O ZJSJQWDXAYNLNS-UHFFFAOYSA-N 0.000 description 3
- ZJSJQWDXAYNLNS-FUPWJLLWSA-N (2s,3r,4s,5s,6r)-2-[4-[(3s,3ar,6s,6ar)-6-[3-methoxy-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]-1,3,3a,4,6,6a-hexahydrofuro[3,4-c]furan-3-yl]-2-methoxyphenoxy]-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound COC1=CC([C@@H]2[C@@H]3[C@@H]([C@H](OC3)C=3C=C(OC)C(O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)=CC=3)CO2)=CC=C1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O ZJSJQWDXAYNLNS-FUPWJLLWSA-N 0.000 description 3
- CWVRJTMFETXNAD-FWCWNIRPSA-N 3-O-Caffeoylquinic acid Natural products O[C@H]1[C@@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-FWCWNIRPSA-N 0.000 description 3
- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 description 3
- 241000235646 Cyberlindnera jadinii Species 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 230000003110 anti-inflammatory effect Effects 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 208000006673 asthma Diseases 0.000 description 3
- 230000036772 blood pressure Effects 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 description 3
- 235000001368 chlorogenic acid Nutrition 0.000 description 3
- 229940074393 chlorogenic acid Drugs 0.000 description 3
- FFQSDFBBSXGVKF-KHSQJDLVSA-N chlorogenic acid Natural products O[C@@H]1C[C@](O)(C[C@@H](CC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O FFQSDFBBSXGVKF-KHSQJDLVSA-N 0.000 description 3
- BMRSEYFENKXDIS-KLZCAUPSSA-N cis-3-O-p-coumaroylquinic acid Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)cc2)[C@@H]1O)C(=O)O BMRSEYFENKXDIS-KLZCAUPSSA-N 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 238000013329 compounding Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 description 3
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 230000000529 probiotic effect Effects 0.000 description 3
- 240000006439 Aspergillus oryzae Species 0.000 description 2
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 2
- 101710130006 Beta-glucanase Proteins 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 2
- 235000019750 Crude protein Nutrition 0.000 description 2
- 108010059378 Endopeptidases Proteins 0.000 description 2
- 102000005593 Endopeptidases Human genes 0.000 description 2
- IBFYXTRXDNAPMM-BVTMAQQCSA-N Geniposide Chemical compound O([C@@H]1OC=C([C@@H]2[C@H]1C(=CC2)CO)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O IBFYXTRXDNAPMM-BVTMAQQCSA-N 0.000 description 2
- IBFYXTRXDNAPMM-FZEIBHLUSA-N Geniposide Natural products COC(=O)C1=CO[C@@H](O[C@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2O)[C@H]2[C@@H]1CC=C2CO IBFYXTRXDNAPMM-FZEIBHLUSA-N 0.000 description 2
- 206010062717 Increased upper airway secretion Diseases 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 241000235342 Saccharomycetes Species 0.000 description 2
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 2
- 241001148470 aerobic bacillus Species 0.000 description 2
- 230000003712 anti-aging effect Effects 0.000 description 2
- 230000001088 anti-asthma Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- VGLLGNISLBPZNL-RBUKDIBWSA-N arborescoside Natural products O=C(OC)C=1[C@@H]2C([C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)OC=1)=C(CO)CC2 VGLLGNISLBPZNL-RBUKDIBWSA-N 0.000 description 2
- 229940107666 astragalus root Drugs 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229940126678 chinese medicines Drugs 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 2
- 229940066758 endopeptidases Drugs 0.000 description 2
- 238000002481 ethanol extraction Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229920005610 lignin Polymers 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 208000026435 phlegm Diseases 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- 210000004927 skin cell Anatomy 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000004575 stone Substances 0.000 description 2
- 239000000052 vinegar Substances 0.000 description 2
- 235000021419 vinegar Nutrition 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- OWVNHBRJANEEKY-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid;hydrochloride Chemical compound Cl.OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O OWVNHBRJANEEKY-UHFFFAOYSA-N 0.000 description 1
- 108091005508 Acid proteases Proteins 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000205585 Aquilegia canadensis Species 0.000 description 1
- 108091005658 Basic proteases Proteins 0.000 description 1
- 235000019224 Camellia sinensis var Qingmao Nutrition 0.000 description 1
- 241000007126 Codonopsis pilosula Species 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 102000018389 Exopeptidases Human genes 0.000 description 1
- 108010091443 Exopeptidases Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000589236 Gluconobacter Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102000035092 Neutral proteases Human genes 0.000 description 1
- 108091005507 Neutral proteases Proteins 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 241000606860 Pasteurella Species 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 206010051246 Photodermatosis Diseases 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 206010040880 Skin irritation Diseases 0.000 description 1
- 206010040954 Skin wrinkling Diseases 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 235000002597 Solanum melongena Nutrition 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000000954 anitussive effect Effects 0.000 description 1
- 230000002391 anti-complement effect Effects 0.000 description 1
- 239000000924 antiasthmatic agent Substances 0.000 description 1
- 239000002220 antihypertensive agent Substances 0.000 description 1
- 229940127088 antihypertensive drug Drugs 0.000 description 1
- 229940124584 antitussives Drugs 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 235000013405 beer Nutrition 0.000 description 1
- 108010047754 beta-Glucosidase Proteins 0.000 description 1
- 102000006995 beta-Glucosidase Human genes 0.000 description 1
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 1
- 235000013734 beta-carotene Nutrition 0.000 description 1
- 239000011648 beta-carotene Substances 0.000 description 1
- 229960002747 betacarotene Drugs 0.000 description 1
- 239000011942 biocatalyst Substances 0.000 description 1
- 238000010364 biochemical engineering Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 230000004154 complement system Effects 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- PCHPORCSPXIHLZ-UHFFFAOYSA-N diphenhydramine hydrochloride Chemical compound [Cl-].C=1C=CC=CC=1C(OCC[NH+](C)C)C1=CC=CC=C1 PCHPORCSPXIHLZ-UHFFFAOYSA-N 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 210000003746 feather Anatomy 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 235000019990 fruit wine Nutrition 0.000 description 1
- 235000009569 green tea Nutrition 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 125000000468 ketone group Chemical group 0.000 description 1
- 150000002576 ketones Chemical group 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 229930013686 lignan Natural products 0.000 description 1
- 235000009408 lignans Nutrition 0.000 description 1
- 150000005692 lignans Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 108020004410 pectinesterase Proteins 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229930015704 phenylpropanoid Natural products 0.000 description 1
- 125000001474 phenylpropanoid group Chemical group 0.000 description 1
- 230000008845 photoaging Effects 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000011533 pre-incubation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 235000020339 pu-erh tea Nutrition 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 229940079877 pyrogallol Drugs 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 230000009759 skin aging Effects 0.000 description 1
- 230000036556 skin irritation Effects 0.000 description 1
- 231100000475 skin irritation Toxicity 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000009044 synergistic interaction Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Birds (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a preparation method of eucommia ulmoides fermentation extracting solution, the eucommia ulmoides fermentation extracting solution prepared by the method and application of the eucommia ulmoides fermentation extracting solution to preparation of cosmetics. The preparation method comprises the following steps: a step of preparing a composite strain, in which acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae are pre-cultured respectively to obtain the composite strain; the method comprises the steps of performing enzymolysis on eucommia ulmoides powder to prepare a fermentation substrate, wherein in the step, eucommia ulmoides leaves are cleaned, dried and crushed into powder, water is added to the powder to be uniformly mixed, then protease, cellulase and pectinase are added to the powder for enzymolysis, so that an enzymolysis liquid of the eucommia ulmoides is obtained, and the fermentation substrate is prepared on the basis of the enzymolysis liquid; a step of fermentation, in which the composite strain is inoculated into a fermentation substrate containing the enzymolyzed eucommia ulmoides enzymolysis liquid, and fermentation is carried out under the condition of low dissolved oxygen to obtain fermentation liquid; and a purification step of centrifuging and filtering the fermentation liquid to obtain eucommia ulmoides fermentation extract.
Description
Technical Field
The invention belongs to the field of biochemical engineering, and particularly relates to preparation of an eucommia ulmoides fermentation extracting solution and application of the eucommia ulmoides fermentation extracting solution in cosmetics.
Background
In China, the application of plant Chinese medicines to natural skin care products has been in history for thousands of years, and compared with chemical composites, the plant Chinese medicines have the characteristics of good efficacy, small side effect and higher safety, so in recent years, skin care products mainly comprising pure natural plant components are increasingly popular with consumers. Eucommia ulmoides (Eucomiaauloides Oliver) is a unique and precious Chinese medicinal material in China, has a medicinal history of more than 2000 years, is called as 'activated stone plants', is distributed in more than 20 provinces in China, and currently, the planting area of the eucommia ulmoides in China has been developed to 35 ten thousand hectares and accounts for more than 99 percent of the total amount of the eucommia ulmoides in the world. Eucommia bark is rich in various active ingredients such as chlorogenic acid, polysaccharide, flavone and the like, and modern pharmacological studies prove that: the folium Eucommiae and cortex Eucommiae have similar pharmacological effects, and have effects of strengthening tendons and bones, lowering blood pressure and blood lipid, inhibiting bacteria, relieving inflammation, resisting oxidation, resisting aging, enhancing immunity, inducing diuresis, and clearing heat.
Eucommia fermentation often relates to the field of food health products, and the field of cosmetics relates to a small amount. In the field of cosmetics, eucommia ulmoides has been studied mainly from eucommia ulmoides extracts obtained by physical or chemical methods. Chinese patent CN102885726A discloses an eucommia ulmoides extract, a preparation method and application thereof, wherein the eucommia ulmoides extract is obtained by an extraction and recovery method of organic solvents such as isopropanol, methanol, acetone, ethyl acetate and the like, and has the effects of oxidation resistance and photoaging resistance. Chinese patent CN105616273A discloses the application of red eucommia ulmoides and its extract in preparing anti-aging skin care products, wherein the eucommia ulmoides extract is obtained by an ethanol extraction method, and has anti-aging effect. The disadvantages of the above method mainly lie in: (1) physical or chemical extraction method, the extraction rate of active ingredients is not high, and the extraction process has residues of organic reagents such as acetone, ethyl acetate and the like, which may not be beneficial to skin and human body and may cause pollution to the environment; (2) more than 80 active ingredients in eucommia bark are mainly flavonoid, polysaccharide, lignin, phenylpropanoids and the like, and the active ingredients cannot be effectively exerted by only once development and utilization of eucommia bark raw materials by a physical or chemical extraction method, so that the waste of traditional Chinese medicine resources is caused.
Chinese patent CN107927497A discloses a "method for producing a fermented beverage of eucommia bark", which is characterized in that the fermented beverage of eucommia bark is obtained by leaching eucommia leaves, enzymolysis, primary fermentation of lactic acid bacteria and secondary fermentation of yeast at 4-6 ℃, and the defects are that: the optimal growth temperature of the yeast is 25-30 ℃, and the fermentation at 4-6 ℃ is not beneficial to the growth metabolic activity of the yeast and the catalytic action of enzyme, thereby being not beneficial to the biological transformation of effective components in the eucommia leaves. The sugar degree of the fermented beverage is 7% -20%, the fermented beverage contains a large amount of live lactic acid bacteria and saccharomycetes, the obtained fermentation liquor is not subjected to subsequent treatment, the product is mainly applied to the field of food and health care products, the product is difficult to be applied to a formula as a cosmetic raw material, and if the product is directly used, a microecological balance system on the surface of skin can be damaged.
Chinese patent CN106010926A discloses a eucommia ulmoides fermentation health-preserving vinegar and a preparation method thereof, which is characterized in that firstly, an eucommia ulmoides extracting solution is obtained through enzymolysis and ethanol extraction methods, secondly, a fruit wine is prepared through primary fermentation of saccharomycetes, and a eucommia ulmoides fermentation health-preserving vinegar is obtained through secondary fermentation of acetic acid bacteria, wherein the main defects are as follows: the fermentation period is long, the production efficiency is low, the product contains a large amount of viable bacteria, acetic acid and alcohol substances, the product is mainly applied to the field of food health products, the product cannot be directly applied as a cosmetic raw material, and if the product is directly used, skin irritation is easily caused.
Chinese patent CN109498744A discloses a method for preparing compound eucommia leaf powder by compound strain fermentation, which is characterized in that a traditional Chinese medicine microecological preparation is obtained by compound fermentation of bacillus subtilis, saccharomyces cerevisiae and astragalus root nodule bacteria and a traditional Chinese medicine culture medium containing astragalus root, codonopsis pilosula, sealwort, honeysuckle, eucommia leaves and the like: the compound eucommia bark leaf powder has the main disadvantages that: the long period of solid fermentation leads to low production efficiency, the culture process is easy to be infected with mixed bacteria, the fermentation strain is not probiotic bacteria, the components of the culture medium are expensive, and the solid fermentation culture medium is mainly applied to the field of feeds and can not be directly used as cosmetic raw materials.
Chinese patent CN108721367A discloses the application of a compound zymophyte in preparation of eucommia ulmoides decoction pieces, which is characterized in that the eucommia ulmoides decoction pieces are obtained by slicing compound zymophyte eucommia ulmoides bark of aspergillus oryzae, bacillus natto and candida utilis and drying at low temperature, and the defects are that: the fermentation strain is not a probiotic, and although the contents of chlorogenic acid and pinoresinol diglucoside in the eucommia ulmoides slices are improved through fermentation, the active ingredients are difficult to release into the fermentation liquor due to the fermentation of the eucommia ulmoides slices, so that the fermentation strain is mainly applied to the field of food health products and cannot be directly applied as cosmetic raw materials.
In summary, the application of the extract developed by taking eucommia ulmoides as a raw material in the field of cosmetics mainly has the following defects:
1. eucommia fermentation is mostly applied to the field of food health products, and the fermentation extract of the eucommia fermentation is not reported in application patents in the field of cosmetics.
2. The application of eucommia ulmoides as a raw material in the field of cosmetics mainly obtains an eucommia ulmoides extract by a common extraction method, and has the problems of low extraction rate of active ingredients, organic reagent residue and simplicity in development and utilization of eucommia ulmoides.
3. The patent technology taking eucommia fermentation liquor as a product mainly comprises the step of sequentially fermenting single or multiple probiotics, and does not fully utilize the complementary synergistic advantages of different bacterial strains, thereby causing waste of eucommia resources.
Disclosure of Invention
In order to solve the above problems, the present application provides:
1. a preparation method of eucommia ulmoides fermentation extracting solution comprises the following steps:
a step of preparing a composite strain, in which acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae are pre-cultured respectively to obtain the composite strain;
the method comprises the steps of performing enzymolysis on eucommia ulmoides powder to prepare a fermentation substrate, wherein in the step, eucommia ulmoides leaves are cleaned, dried and crushed into powder, water is added to the powder to be uniformly mixed, then protease, cellulase and pectinase are added to the powder for enzymolysis, so that an enzymolysis liquid of the eucommia ulmoides is obtained, and the fermentation substrate is prepared on the basis of the enzymolysis liquid;
a step of fermentation, in which the composite strain is inoculated into a fermentation substrate containing the enzymolyzed eucommia ulmoides enzymolysis liquid in a total inoculation amount of 5-10% of the volume of the fermentation substrate, and fermentation is carried out under a low dissolved oxygen condition to obtain fermentation liquid;
and a purification step of centrifuging and filtering the fermentation liquid to obtain eucommia ulmoides fermentation extract.
2. The method according to item 1, wherein, in the step of preparing the composite bacterial species, the pre-culture solution of the three bacterial species is obtained by pre-culturing the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae respectively, and in order to ferment by using the composite bacterial species, the volume ratio of the pre-culture solution of the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae used in the subsequent fermentation process is as follows: 1: 1-6: 0.6 to 4;
wherein, in the pre-culture solution, the bacteria concentration of the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae meets the following requirements:
the bacteria concentration in the pre-culture solution of Acetobacter pasteurianus is 1 × 107More than cfu/mL, and the like,
the concentration of the Lactobacillus plantarum preculture solution was 8 × 108More than cfu/mL, and the like,
the concentration of the bacteria in the pre-culture solution of the saccharomyces cerevisiae is 3 × 108cfu/mL or more.
3. The method of item 1, wherein, in the step of enzymatically hydrolyzing the eucommia ulmoides oliv powder to prepare the fermentation substrate, the mass fraction of the eucommia ulmoides oliv leaves is 5% to 15%, the addition amount of the protease is 0.1% to 0.5% of the weight of the selected eucommia ulmoides oliv leaves, the addition amount of the cellulase is 0.2% to 1.0% of the weight of the selected eucommia ulmoides oliv leaves, the addition amount of the pectinase is 0.1% to 1.0% of the weight of the selected eucommia ulmoides oliv leaves, and the specific activities of the enzymes are not less than 3000U/g.
4. The method of item 1, wherein, in the step of enzymatically hydrolyzing the eucommia ulmoides powder to prepare the fermentation substrate, the enzymatic hydrolysis reaction conditions are: the pH value is 3.5-6.5, the temperature is 45-60 ℃, the enzymolysis time is 1-3.5 h, and when the enzymolysis is finished, the mixture is boiled for 3-5 min to inactivate the enzyme.
5. The method according to item 1, wherein the fermentation substrate comprises, in addition to the eucommia ulmoides enzymolysis solution: a nitrogen source, a carbon source, inorganic salt and the balance of drinking water, wherein the nitrogen source is selected from any one of yeast powder, tryptone, soy peptone or beef extract; the carbon source is selected from any one of sucrose, glucose, lactose or galactose; the inorganic salt is selected from NaCl and K2HPO4。
6. The method according to item 5, wherein the nitrogen source is 1 to 5 parts, the carbon source is 1 to 5 parts, NaCl is 0.1 to 0.5 part, and K is added to 100 parts by weight of the fermentation substrate2HPO40.1 to 0.5 portion.
7. The method according to item 1, wherein, in the step of performing fermentation, the fermentation temperature is 30 ℃ to 35 ℃, acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae are inoculated either sequentially or simultaneously, preferably simultaneously, the pH of the fermentation substrate is adjusted to 6.0 to 7.0, the fermentation time is 36h to 48h, the dissolved oxygen during the culture process is controlled to 10% to 40%, and the end point of fermentation is judged as the end point if no residual carbon source exists.
8. The method of item 1, wherein, in the purification, the centrifugation conditions are set as: cells were removed by centrifugation at 9000rpm for at least 20 minutes.
9. The method of item 1, wherein, in the purification, the filtration conditions are set as: adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core.
10. A fermented extract of eucommia ulmoides prepared by the method according to any one of items 1 to 9.
11. Application of fermented extract of Eucommiae cortex in preparing cosmetic is provided.
The technical scheme of the application obtains the following beneficial effects:
1. the invention adopts a preparation method of multi-enzymolysis and probiotic composite fermentation, so that the obtained eucommia fermentation extract has better effects, such as good oxidation resistance and anti-inflammatory performance. Therefore, the eucommia ulmoides oliv extract has wide application potential in the field of cosmetics, and the application field of the eucommia ulmoides oliv is fully developed.
2. The eucommia ulmoides sources in China are wide and easy to obtain, the eucommia ulmoides sources are low in price, and a microbial fermentation method is green and has little pollution; compared with the eucommia bark extract obtained by a physical and chemical method, the eucommia bark fermented extract obtained by the invention retains the active ingredients to a greater extent, and has good efficacy, small side effect and higher safety.
3. According to the invention, three probiotics are selected to carry out synergistic fermentation on the folium cortex eucommiae enzymolysis substrate, so that the operation procedures are reduced, the production cost is reduced, different growth characteristics and synergistic interaction of the three probiotics are fully utilized, and the folium cortex eucommiae is fully developed and utilized.
Detailed Description
The technical solution of the present application is explained in detail below.
< eucommia ulmoides >
Eucommia ulmoides (eucommia ulmoides Oliver) is a unique plant 'activated stone' in China, only belongs to one family, is a perennial deciduous tree plant of eucommia of the family eucommia, and has been living on the earth for nearly 5000 ten thousand years. The eucommia ulmoides tree species has high economic value and rare resources, belongs to national-grade rare or endangered plants, and is particularly classified as national-grade second-grade precious protection tree species by relevant departments of the government of China in order to protect eucommia ulmoides resources. At least two thousand years ago, the eucommia ulmoides is tried by the people to make the body strong and prolong the life. Eucommia bark is a treasure on the whole body, and eucommia seed, bark, leaf, flower and branch are developed and utilized in many aspects. Because eucommia bark is a medicinal plant, the secondary metabolite of the eucommia bark has great utilization value for human beings, and the secondary metabolite of the eucommia bark mainly comprises lignin, iridoid, flavonoid, phenol, triterpenes, steroids, polysaccharide, organic acid, amino acid, trace elements and the like.
Application of eucommia ulmoides in medical treatment
1.1 the eucommia bark has the function of reducing blood pressure
Eucommia ulmoides is regarded as the highest quality natural antihypertensive drug without side effects in the world at present, and the effective component for lowering blood pressure is pinoresinol diglucoside.
1.2 the effect of eucommia bark on improving immunity
Through literature data analysis, the eucommia ulmoides leaf extract preparation is found to have better promotion effect on the immunity function of mice than cordyceps sinensis compared with the acknowledged cordyceps sinensis extract preparation for improving the immunity of human bodies and animals.
1.3 the effects of relieving cough and asthma and eliminating phlegm of eucommia bark
Eucommia bark contains a plurality of flavonoid compounds, and clinical experiments in recent decades show that the eucommia bark has the effects of relieving cough and asthma and eliminating phlegm. It is reported that the antiasthmatic action of flavonoids is related to the unsaturated ketone structure in the molecular structure, and the stronger the oxygen nucleophilic ability of ketone group, the stronger the antitussive, antiasthmatic and expectorant actions.
1.4 the anti-complement effect of eucommia ulmoides
The complement system is one of important immune components, plays an important role in the regulation of defense function and immune function of the body, but abnormal activation can cause serious pathological damage to human body self tissues, such as rheumatoid arthritis, asthma and other diseases related to autoimmune runaway, and the change of the complement content is often used as an important detection index for the diseases. Therefore, the related diseases can be treated by screening out the drugs for inhibiting the excessive activation of the complement.
1.5 anti-tumor effect of eucommia ulmoides
The effective components of eucommia bark with anti-tumor effect are various lignans, geniposide and geniposide, and the anti-tumor effect of eucommia bark is possibly related to β -carotene.
Application of eucommia ulmoides in health care
2.1 the eucommia bark has the function of losing weight
Human body tests prove that all people taking the eucommia tea continuously for more than one month can obviously reduce the content of the neutral fat under the skin and around the internal organs of the human body, play the roles of no exercise, no change of diet life, obesity prevention and weight loss. The eucommia tea can promote the synthesis of protein in a human body, promote metabolism, consume energy in the body and naturally reduce neutral fat accumulated in the body, so the eucommia tea can have the weight-losing effect after being drunk for a long time.
2.2 the cortex Eucommiae has skin caring effect
Human skin aging is mainly caused by the loss of elasticity of dermal cells, the collagen in the dermal cells accounts for 70%, and the eucommia ulmoides can accelerate the metabolism of the collagen and restore the elasticity, so that the beauty effect of preventing skin wrinkling is achieved.
2.3 health promotion effect of eucommia ulmoides tea
Eucommia ulmoides tea is developed by using eucommia ulmoides leaves, and health care tea is developed by using the eucommia ulmoides leaves in Guizhou, Sichuan and the like in the early stage of the 80 s. The fan, the English longevity and the like invent the folium cortex eucommiae tea with the Pu' er tea style; the feather xi extracts a plurality of traditional Chinese medicine juices such as eucommia bark and the like, and the juice is added into green tea to develop the sanjia shirt eucommia bark tea. A great deal of research is carried out on the health care efficacy of eucommia ulmoides tea in Japan, and animal experiments show that: can reduce cholesterol and neutral lipid; the artificial eel is fed with 2.5% eucommia tea soup by Gaoqiao to reduce the sexual fat by 20%, and the muscle tissue is compact and the elasticity is increased; the eucommia ulmoides tea can improve the laying rate of the laying hens by 20 percent in field tests. Therefore, the development of the health-care tea by utilizing the eucommia ulmoides leaf resources has great advantages in aspects such as the utilization value of the leaves, market development and the like.
The technical scheme of the application utilizes the beautifying function of eucommia ulmoides and related metabolites thereof.
< enzymatic hydrolysis Process >
The enzyme is a biocatalyst. In the actual production process of extracting natural components, enzymolysis reaction is often used, because some enzymes can decompose plant cell walls under normal temperature, normal pressure and mild acid-base conditions, the extraction rate of effective components in natural plants can be greatly improved, and the product purity and the preparation quality are improved.
Commonly used enzymes include enzymes for breaking cell walls of plant cells and enzymes for separating, refining and improving the clarity of extraction; the former is cellulase and pectinase, and the latter is papain.
These two classes of enzymes are used in combination in the protocol of the present application.
"composite use" herein encompasses both sequential and simultaneous use. The compound use mode comprises the following steps:
firstly, protease, cellulase and pectinase are used;
firstly, protease, pectinase and cellulase are used;
firstly, pectinase, protease and cellulase are used;
firstly, pectinase, cellulase and protease are used;
firstly, cellulase, pectinase and protease are used;
firstly, cellulase, then protease and finally pectinase are used;
firstly, protease and cellulase are used in a compounding way, and then pectinase is used;
firstly, protease and pectinase are used in a compounding manner, and then cellulase is used;
firstly, pectinase and cellulase are used in a compounding manner, and then protease is used;
firstly, cellulase is used, and then protease and pectinase are simultaneously used;
firstly, protease is used, and then cellulase and pectinase are used simultaneously;
firstly, pectinase is used, and then protease and cellulase are used simultaneously;
protease, pectinase and cellulase are used simultaneously.
In the context of the present application, proteases are a general term for enzymes that hydrolyze peptide chains of proteins. By their way of hydrolyzing polypeptides, they can be divided into two classes, endopeptidases and exopeptidases, in which, for example, the most frequently used industrially endopeptidases are used; depending on the pH optimum, acid proteases, neutral proteases and alkaline proteases can be used, for example.
In the context of the present application, pectinase is a generic term for enzymes that break down pectic substances. The reaction mechanism can be divided into two types, one can catalyze depolymerization of pectin, and the other can catalyze hydrolysis of ester in pectin molecule. Wherein, the former can be divided into enzymes acting on pectin and enzymes acting on pectic acid; the latter may be classified into pectinesterase and pectoyl hydrolase. For example, enzymes acting on pectin are used here.
In the context of the present application, cellulase is a generic term for enzymes that degrade cellulose to form glucose, is not a monomeric enzyme, but a synergistic multi-component enzyme system, is a complex enzyme, mainly consisting of exo- β -glucanase, endo- β -glucanase, β -glucosidase, etc., which acts on cellulose and products derived from cellulose.
In the context of the present application, "enzymatic activity" complies with the conventional definition in enzymology, i.e. is an indication of the ability of an enzyme to catalyze a certain chemical reaction, in particular the conversion rate of the certain chemical reaction it catalyzes, i.e. the faster the conversion rate catalyzed by the enzyme, the higher the enzymatic activity; conversely, the slower the rate, the less active the enzyme. Therefore, the determination of the enzyme activity is the determination of the enzymatic conversion rate. The enzymatic conversion rate can be expressed in terms of the decrease in substrate per unit volume or the increase in product per unit time. "specific activity" means the enzymatic activity of an enzyme protein per unit mass, and usually, the unit is U/mg, and may be U/g. The higher the specific activity, the purer the enzyme. In the technical scheme of the application, the specific activity of each enzyme is not lower than 3000U/g.
The common influencing factors of the enzymolysis reaction are temperature, pH value, coenzyme supply and the like. Enzymolysis reaction phaseA particular advantage of the general chemical reactions is that the catalytic conditions of the enzymatic reactions are generally mild and can be carried out at normal temperature and pressure 10 times faster than the corresponding non-catalytic reactions3-107And (4) doubling. In the technical scheme of the application, the enzymolysis reaction conditions are as follows: the pH value is 3.5-6.5, and the temperature is 45-60 ℃.
At the end of the enzymatic hydrolysis process, the enzymatic reaction may be terminated by deactivating the enzyme by boiling. The end point of the enzymatic reaction can be determined by measuring the glucose and crude protein content, which in the context of this application is defined as the time interval between 0.5h when the glucose and crude protein content does not change.
< fermentation Process >
According to the requirement of microorganism on oxygen in the fermentation process, the fermentation types are mainly as follows: nutrient fermentation, anaerobic fermentation and facultative anaerobic fermentation are required. According to the technical scheme, the method comprises acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae. In the culture process, the fermentation is carried out by adopting a low dissolved oxygen culture mode mainly according to the growth characteristics of the three bacteria, and under a proper combination, the different bacteria can play a synergistic effect in the co-culture, so that the culture process is simplified, the operation procedures are reduced, and the synergistic effect is fully utilized. The application utilizes the synergistic fermentation of three bacteria: acetobacter pasteurianus, Lactobacillus plantarum and Saccharomyces cerevisiae.
In the application, the acetobacter pasteurianus is aerobic bacteria, and the lactobacillus plantarum and the saccharomyces cerevisiae are facultative anaerobic bacteria, so that a low dissolved oxygen fermentation mode is used for the composite fermentation of the three bacteria.
The term "synergistic fermentation" herein covers fermentation modes of sequential inoculation and simultaneous inoculation, including:
inoculating acetobacter pasteurianus, then inoculating lactobacillus plantarum, and finally inoculating saccharomyces cerevisiae;
inoculating acetobacter pasteurianus, then inoculating saccharomyces cerevisiae, and finally inoculating lactobacillus plantarum;
inoculating saccharomyces cerevisiae, then inoculating acetobacter pasteurianus and finally inoculating lactobacillus plantarum;
inoculating saccharomyces cerevisiae, then inoculating lactobacillus plantarum, and finally inoculating acetobacter pasteurianus;
inoculating lactobacillus plantarum, then inoculating saccharomyces cerevisiae, and finally inoculating acetobacter pasteurianus;
inoculating lactobacillus plantarum, then inoculating acetobacter pasteurianus and finally inoculating saccharomyces cerevisiae;
firstly, simultaneously inoculating acetobacter pasteurianus and lactobacillus plantarum, and then inoculating saccharomyces cerevisiae;
firstly, simultaneously inoculating acetobacter pasteurianus and saccharomyces cerevisiae, and then inoculating lactobacillus plantarum;
simultaneously inoculating saccharomyces cerevisiae and lactobacillus plantarum, and then inoculating acetobacter pasteurianus;
inoculating saccharomyces cerevisiae, and then inoculating lactobacillus plantarum and acetobacter pasteurianus at the same time;
inoculating lactobacillus plantarum, and then inoculating saccharomyces cerevisiae and acetobacter pasteurianus at the same time;
inoculating acetobacter pasteurianus, and then inoculating lactobacillus plantarum and saccharomyces cerevisiae simultaneously;
simultaneously inoculating acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae.
When the fermentation substrate is sequentially inoculated according to the above steps, the total inoculation amount of the strains is 5-10% of the volume of the fermentation substrate.
For the pre-culture solution, the bacteria concentration of the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae meets the following conditions:
the bacteria concentration in the pre-culture solution of Acetobacter pasteurianus is 1 × 107More than cfu/mL, and the like,
the concentration of the Lactobacillus plantarum preculture solution was 8 × 108More than cfu/mL, and the like,
the concentration of the bacteria in the pre-culture solution of the saccharomyces cerevisiae is 3 × 108cfu/mL or more.
In the context of this application, saccharomyces is a unicellular fungus that ferments sugars to ethanol and carbon dioxide, distributed throughout nature, a typical heterotrophic facultative anaerobic microorganism that is viable under both aerobic and anaerobic conditions, saccharomyces cerevisiae being an important class of yeast.
In the context of the present application, acetic acid bacteria are one of the important industrial bacteria, gram-negative, aerobic bacteria, capable of oxidizing organic matter to organic acids. The important characteristic of acetic acid bacteria is that they can oxidize ethanol into acetic acid, and they are classified into two major categories according to their optimum development temperature and characteristics, one is called acetobacter and the other is called gluconobacter, the pasteurella in this application is acetobacter. The metabolic type of acetic acid bacteria belongs to heterotrophic aerobic type, so oxygen is always required to participate in fermentation.
In the context of the present application, lactic acid bacteria are a general term for a group of bacteria that are capable of producing large amounts of lactic acid using fermentable carbohydrates. Lactic acid bacteria, gram-positive, frequently arranged in chains; according to different sources, the lactobacillus can be divided into animal source lactobacillus and plant source lactobacillus, and the lactobacillus plantarum in the application belongs to the plant source lactobacillus; metabolically, the genus lactobacillus is a heterotrophic facultative anaerobic microorganism.
Therefore, in the technical scheme of the application, in order to realize the synergistic fermentation of the three bacteria, the dissolved oxygen is controlled to be 10-40% (low dissolved oxygen).
The individual species need to be pre-cultured before the composite species are inoculated simultaneously or sequentially into the fermentation substrate.
In the context of the present application, "pre-culture" is in accordance with the general definition in the art, and specifically refers to a process of obtaining a pure culture of a certain quantity and quality by inoculating a production strain in a dormant state in a sand tube or a freeze-drying tube into a test tube slant for activation, and then performing stepwise scale-up culture in an eggplant bottle or a shake flask and a seed tank. These true cultures are also referred to as seed solutions or precultures.
The seeds in the fermentation industrial production process must meet the following conditions:
(1) the growth activity of the strain cells is strong, and the strain cells can grow rapidly after being transplanted to a fermentation tank;
(2) the physiological shape is stable;
(3) the total amount and concentration of the thalli can meet the requirements of a large-capacity fermentation tank;
(4) 2, no mixed bacteria pollution is caused;
(5) maintaining a stable production capacity.
Here, the seed preparation process of Saccharomyces cerevisiae is only taken as an example:
the process is briefly described as follows: test tube → triangular flask → seeding tank
Taking the general beer-producing Saccharomyces cerevisiae as an example: it is generally stored on a wort agar slant and stored in a refrigerator at 4 ℃. Inoculating the preserved Saccharomyces cerevisiae strain into 500-1000 mL triangular flask containing 100mL wort, culturing at 25 deg.C for 2-3 days, expanding to 500-1000 mL triangular flask containing 250-500 mL wort, culturing at 25 deg.C for 2 days, transferring to seed tank containing 5-10L wort, and culturing at 15-20 deg.C for 3-5 days to obtain 100L wort seed solution.
Here, for example, for 100L of fermentation substrate, 5L (5%) of yeast preculture are required for the inoculation. Of course, the pre-incubation shown herein is for illustration purposes only and is not limiting.
In the context of the present application, "low dissolved oxygen" complies with the general definition in the field of fermentation engineering. "dissolved oxygen" (DO) refers to oxygen in the molecular state dissolved in water. The principle of controlling dissolved oxygen consists in increasing the oxygen transfer rate, the usual operating variables being temperature, pressure, air volume and the rotational speed of the stirring paddles (stirring power). In the technical scheme of the application, the factors are controlled to enable the DO value to be 10% -40% so as to meet the requirement of fermentation under the condition of low dissolved oxygen.
< purification Process >
Purification refers to the process by which aggregates of multiple substances are changed into one class or substance by physical, chemical or biological means. Purification can be a general concept, including processes that can separate and purify all mixtures, such as distillation, extraction, crystallization, and the like. In the context of the present application, "purification" refers in particular to the process of separating a fermentation extract from a fermentation broth by various technical means.
< examples section >
The present invention is further described below with reference to specific examples so that those skilled in the art can more easily understand the present invention, but the present invention is not limited thereto.
Example 1 preparation of fermented extract of eucommia ulmoides
(1) The preparation method of the composite strain comprises the steps of respectively inoculating acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae into different triangular flasks from inclined planes, standing and culturing for 48 hours for expansion culture, wherein the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae are composed according to the ratio of 1:2:0.6 to obtain the composite strain, and the viable bacteria number of each strain in the pre-culture solution is respectively that the acetobacter pasteurianus is not less than 1 × 107cfu/mL, Lactobacillus plantarum no less than 8 × 108cfu/mL, Saccharomyces cerevisiae not less than 3 × 108cfu/mL。
(2) Preparation of eucommia fermentation substrate: taking 10kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 9.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder in 100L of drinking water, adjusting the pH to 5.0 by using hydrochloric acid, wherein the addition amount of protease is 0.3 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 0.6 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 0.5 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 50 ℃ for 2.5 hours, and then boiling for 3-5 min to inactivate the enzymes.
(3) Fermentation: adding 2.5kg glucose, 3.0kg yeast powder, 0.3kg sodium chloride, and 0.25kg K into above Eucommiae cortex zymolysis substrate2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 32 ℃, inoculating 7.5% of composite strain seed liquid according to the volume ratio, culturing for 42h at 32 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquid to be 6.0-7.0, and ending the fermentation after no residual sugar exists.
(4) And (3) purification: centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A1.
Example 2 preparation of fermented extract of eucommia ulmoides
(1) Preparing a composite strain: inoculating Acetobacter pasteurianus, Lactobacillus plantarum and Saccharomyces cerevisiae to different triangular flasks from inclined planes respectively, standing and culturing for 48h for amplification cultureThe composite strain is prepared from acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae according to the proportion of 1:3:4, and the number of live bacteria of each strain in the pre-culture solution is respectively that the acetobacter pasteurianus is not less than 1 × 107cfu/mL, Lactobacillus plantarum no less than 8 × 108cfu/mL, Saccharomyces cerevisiae not less than 3 × 108cfu/mL。
(2) Preparation of eucommia fermentation substrate: taking 5kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 4.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder with 100L of drinking water, adjusting the pH to 3.5 by hydrochloric acid, wherein the addition amount of protease is 0.1 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 0.2 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 0.1 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 60 ℃ for 1.0h, and then boiling for 3-5 min to inactivate the enzyme.
(3) Fermentation: to the above-mentioned substrate for the enzymatic hydrolysis of eucommia ulmoides was added 5kg of glucose, 5kg of tryptone, 0.5kg of sodium chloride and 0.5kg of K2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 30 ℃, inoculating 10% of composite strain seed liquid according to the volume ratio, culturing for 36h at 30 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquid to be 6.0-7.0, and ending the fermentation after no residual sugar exists.
(4) And (3) purification: centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A2.
Example 3 preparation of fermented extract of eucommia ulmoides
(1) The preparation method of the composite strain comprises the steps of respectively inoculating acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae into different triangular flasks from inclined planes, standing and culturing for 48 hours for expansion culture, wherein the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae are composed according to the ratio of 1:2:1.5 to obtain the composite strain, and the viable bacteria number of each strain in the pre-culture solution is respectively that the acetobacter pasteurianus is not less than 1 × 107cfu/mL, Lactobacillus plantarum no less than 8 × 108cfu/mL, Saccharomyces cerevisiae not less than 3 × 108cfu/mL。
(2) Preparation of eucommia fermentation substrate: taking 15kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 14.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder in 100L of drinking water, adjusting the pH to 6.5 by using hydrochloric acid, wherein the addition amount of protease is 0.1 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 1.0 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 1.0 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 45 ℃ for 3.5 hours, and then boiling for 3-5 minutes to inactivate the enzymes.
(3) Fermentation: adding lactose 1kg, yeast powder 1kg, sodium chloride 0.1kg, and K0.1 kg into the above enzymolysis substrate of Eucommiae cortex2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 35 ℃, inoculating 5% of composite strain seed liquid according to the volume ratio, culturing for 48h at 35 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquid to be 6.0-7.0, and ending the fermentation after no residual sugar exists.
(4) And (3) purification: centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A3.
Example 4 preparation of fermented extract of eucommia ulmoides
(1) The preparation method of the composite strain comprises the steps of respectively inoculating acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae into different triangular flasks from inclined planes, standing and culturing for 48 hours for expansion culture, wherein the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae are composed according to the ratio of 1:1:1.5 to obtain the composite strain, and the viable bacteria number of each strain in the pre-culture solution is respectively that the acetobacter pasteurianus is not less than 1 × 107cfu/mL, Lactobacillus plantarum no less than 8 × 108cfu/mL, Saccharomyces cerevisiae not less than 3 × 108cfu/mL。
(2) Preparation of eucommia fermentation substrate: taking 8kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 7.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder in 100L of drinking water, adjusting the pH to 3.5 by using hydrochloric acid, wherein the addition amount of protease is 0.5 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 0.2 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 0.1 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 45 ℃ for 1.0h, and then boiling for 3-5 min to inactivate the enzymes.
(3) Fermentation: adding 5kg sucrose, 1kg soybean peptone, 0.35kg sodium chloride, and 0.3kg K into above Eucommiae cortex enzymolysis substrate2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 30 ℃, inoculating 9% of composite strain seed liquid according to the volume ratio, culturing for 40h at 35 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquid to be 6.0-7.0, and ending the fermentation after no residual sugar exists.
(4) And (3) purification: centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A4.
Example 5 preparation of fermented extract of eucommia ulmoides
(1) The preparation method of the composite strain comprises the steps of respectively inoculating acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae into different triangular flasks from inclined planes, standing and culturing for 48 hours for expansion culture, wherein the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae are composed according to the ratio of 1:3:2 to obtain the composite strain, and the viable bacteria number of each strain in the pre-culture solution is respectively that the acetobacter pasteurianus is not less than 1 × 107cfu/mL, Lactobacillus plantarum no less than 8 × 108cfu/mL, Saccharomyces cerevisiae not less than 3 × 108cfu/mL。
(2) Preparation of eucommia fermentation substrate: taking 12kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 11.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder with 100L of drinking water, adjusting the pH to 6.5 by using hydrochloric acid, wherein the addition amount of protease is 0.5 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 0.2 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 1.0 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 60 ℃ for 3.5 hours, and then boiling for 3-5 minutes to inactivate the enzymes.
(3) Fermentation: adding 1kg glucose, 5kg yeast powder, 0.35kg sodium chloride, 0.3kg K into above Eucommiae cortex zymolysis substrate2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 30 ℃, inoculating 5% of composite strain seed liquid according to the volume ratio, culturing for 48h at 32 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquid to be 6.0-7.0, and ending the fermentation after no residual sugar exists.
(4) And (3) purification: centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A5.
Example 6 preparation of fermented extract of eucommia ulmoides
(1) The preparation of the strain comprises inoculating Acetobacter pasteurianus, Lactobacillus plantarum and Saccharomyces cerevisiae from inclined plane into different triangular flasks, standing and culturing for 48h for enlarged culture, wherein the viable count of each strain in the pre-culture solution is respectively that the amount of Acetobacter pasteurianus is not less than 1 × 107cfu/mL, Lactobacillus plantarum no less than 8 × 108cfu/mL, Saccharomyces cerevisiae not less than 3 × 108cfu/mL。
(2) Preparation of eucommia fermentation substrate: taking 10kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 9.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder in 100L of drinking water, adjusting the pH to 5.0 by using hydrochloric acid, wherein the addition amount of protease is 0.3 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 0.6 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 0.5 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 50 ℃ for 2.5 hours, and then boiling for 3-5 min to inactivate the enzymes.
(3) Fermentation: adding 2.5kg glucose, 3.0kg yeast powder, 0.3kg sodium chloride, and 0.25kg K into above Eucommiae cortex zymolysis substrate2HPO4Mixing, sterilizing at 116 deg.C for 20min, and cooling to 32 deg.C. Inoculating 2.5% Lactobacillus plantarum into the fermentation mediumCulturing the liquid, and fermenting for 24h to obtain primary fermentation liquid; inoculating 2.5% of culture solution of saccharomyces cerevisiae into the primary fermentation liquid at 32 ℃, and fermenting for 12h to obtain secondary fermentation liquid; inoculating 2.5% Acetobacter pasteurianus culture solution into the secondary fermentation liquid at 37 ℃, and finishing fermentation for 12 h. The pH is controlled to be 6.0-7.0 in the whole fermentation process, and the DO is controlled to be 10% -40% by controlling the rotating speed and the ventilation quantity.
(4) And (3) purification: centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A6.
Comparative example 1
Taking 10kg of eucommia ulmoides leaves as a raw material, cleaning and drying the eucommia ulmoides leaves, fully crushing the eucommia ulmoides leaves into powder by a crusher to obtain 9.5kg of eucommia ulmoides powder, uniformly mixing the eucommia ulmoides powder in 100L of drinking water, adjusting the pH to 5.0 by using hydrochloric acid, wherein the addition amount of protease is 0.3 percent of the weight of the eucommia ulmoides leaves, the addition amount of cellulase is 0.6 percent of the weight of the eucommia ulmoides leaves, the addition amount of pectinase is 0.5 percent of the weight of the eucommia ulmoides leaves, carrying out enzymolysis at 50 ℃ for 2.5 hours, and then boiling for 3-5 min to inactivate the enzymes. Centrifuging the enzymolysis solution at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex extract A7.
Comparative example 2
A fermentation extract of Eucommiae cortex is prepared by performing enzymolysis and fermentation as in comparative example 1, wherein the fermentation strain comprises only Acetobacter pasteurianus, and the number of viable bacteria is not less than 1 × 107cfu/mL, fermentation conditions: adding 1kg glucose, 5kg yeast powder, 0.35kg sodium chloride, 0.3kg K into above Eucommiae cortex zymolysis substrate2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 30 ℃, inoculating 7.5% of seed liquid according to the volume ratio, culturing for 48h at 32 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquor to be 6.0-7.0, and ending the fermentation after no residual sugar exists. Centrifuging the fermentation liquor, sterilizing to obtain cortex Eucommiae extractAnd (6) taking liquid. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A8.
Comparative example 3
A fermentation extract of Eucommiae cortex is prepared by performing enzymolysis and fermentation as in comparative example 1, wherein the fermentation strain only comprises Lactobacillus plantarum, and the number of viable bacteria is not less than 8 × 108cfu/mL, fermentation conditions: adding 1kg glucose, 5kg yeast powder, 0.35kg sodium chloride, 0.3kg K into above Eucommiae cortex zymolysis substrate2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 30 ℃, inoculating 7.5% of seed liquid according to the volume ratio, culturing for 48h at 32 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquor to be 6.0-7.0, and ending the fermentation after no residual sugar exists. Centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core to obtain Eucommiae cortex fermentation extractive solution A9.
Comparative example 4
A fermentation extract of Eucommiae cortex is prepared by performing enzymolysis and fermentation as in comparative example 1, wherein the fermentation strain comprises Saccharomyces cerevisiae, and viable bacteria number is not less than 3 × 108cfu/mL, fermentation conditions: adding 1kg glucose, 5kg yeast powder, 0.35kg sodium chloride, 0.3kg K into above Eucommiae cortex zymolysis substrate2HPO4After fully and uniformly mixing, sterilizing at 116 ℃ for 20min, cooling to 30 ℃, inoculating 7.5% of seed liquid according to the volume ratio, culturing for 48h at 32 ℃, reducing the dissolved oxygen along with the amplification of thalli and the biotransformation process, controlling the rotation speed and the ventilation quantity in the culturing process to ensure that DO is 10% -40%, controlling the pH of the fermentation liquor to be 6.0-7.0, and ending the fermentation after no residual sugar exists. Centrifuging the fermentation liquor, and sterilizing to obtain cortex Eucommiae extractive solution. Centrifuging the fermentation liquid at 9000rpm for 20min to remove thallus, adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, filtering with 1.2 μm fine filter plate and 0.22 μm fine filter plateAnd (4) carrying out core treatment to obtain eucommia ulmoides fermentation extract A10.
Comparative example 5
A preparation method of Eucommiae cortex fermented extractive solution and its application in cosmetics are provided, wherein the composite fermentation strain is prepared from Aspergillus oryzae, Bacillus natto and Candida utilis at a ratio of 4:3:2, and the number of viable bacteria of the composite strain is respectively not less than 1 × 107cfu/mL, Bacillus natto not less than 5 × 107cfu/mL, Candida utilis of not less than 1 × 108cfu/mL. Inoculating the composite strain into a fermentation medium according to 1.0% of the weight of the fermentation medium, and culturing at 32 ℃ for 13.5h to obtain a fermentation liquid, wherein the culture medium comprises: 10% of glucose, 4% of ammonium sulfate, 3% of dipotassium hydrogen phosphate and the balance of water. Selecting and cleaning high-quality eucommia bark, placing in the fermentation liquor, sealing and fermenting for 15h at 33 ℃, centrifuging the fermentation liquor for 20min at 9000rpm to remove thalli, adding 1% of activated carbon, adsorbing for 1h at 55 ℃, and then passing through a 1.2-micron fine filter plate and a 0.22-micron fine filter element to obtain eucommia bark fermentation extract A11.
Antioxidant activity of eucommia ulmoides extract
The experimental method comprises the following steps: the antioxidant activity of the eucommia ulmoides fermentation extract liquid of each example and the eucommia ulmoides extract liquid of the comparative example is respectively measured, and the specific method comprises the following steps:
DPPH radical clearance (%) determination: adding 3mL of phosphate buffer solution (pH6.8), 4mL of DPPH anhydrous ethanol solution (0.4mmol/L) and 0.2mL of extract to be detected into a test tube with a plug in sequence, mixing uniformly, standing at room temperature for 30min, measuring absorbance at 517nm, and taking anhydrous ethanol as blank control.
In the formula: a1, adding the extract to be tested, and measuring the absorbance after reaction;
a2, adding absolute ethyl alcohol into the extract to be detected, and then measuring the absorbance;
a3: adding DPPH and phosphate buffer solution without the solution to be measured to measure the absorbance.
2. Hydroxyl radical clearance (%) measurement: 1mL of salicylic acid solution (9mmol/L), 1mL of the extract to be tested, 1mL of ferrous sulfate solution (0.9mmol/L) and 1mL of hydrogen peroxide solution (8.8mmol/L) are sequentially added into a test tube with a plug, mixed uniformly, subjected to water bath at 37 ℃ for 60min, and the absorbance at 510nm is measured and is recorded as A1, and the absorbance is measured and recorded as A2 by taking purified water as a control.
3. Superoxide anion radical clearance (%) determination: preparing Tris-HCl-EDTA buffer solution (pH8.2), sequentially adding 0.6mL of extract to be detected, 5.2mL of buffer solution and 0.2mL of pyrogallol solution (0.045mol/L) into a test tube with a plug, uniformly mixing, reacting for 5min, measuring the absorbance value at 325nm, marking as A1, and measuring after reacting for 10min, marking as A2. The procedure was performed as described above with reference to purification, and the results were designated as W1 and W2.
And (4) experimental conclusion: the results are shown in table 1, compared with the comparative example, the eucommia ulmoides fermentation extract liquid of each example has improved DPPH free radical clearance rate, hydroxyl free radical clearance rate and superoxide anion free radical clearance rate, and the eucommia ulmoides fermentation extract liquid has good oxidation resistance. By comparing the data of the example with the data of the comparative example 1, the comparative example 2, the comparative example 3 and the comparative example 4, the synergistic fermentation has more prominent antioxidant capacity than that of a single strain and that of the eucommia ulmoides only by enzymolysis, and shows the synergistic effect of the synergistic fermentation of the three strains. By comparing the data of each example with that of comparative example 5, the DPPH radical clearance is improved by about 1.90 to 3.75 times, the hydroxyl radical clearance is improved by about 1.80 to 2.97 times, and the superoxide anion radical clearance is improved by about 2.35 to 3.21 times, which fully shows the advantages of co-fermenting eucommia leaves with acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae presumably because: by adopting the fermentation mode in the comparative example 5, although the contents of chlorogenic acid and pinoresinol diglucoside in the eucommia ulmoides decoction pieces are improved, the active ingredients are not released into the fermentation liquor much or the eucommia ulmoides slices are directly fermented without enzyme treatment before fermentation, and the release of active ingredients such as flavonoids, polysaccharides and the like is less.
TABLE 1 antioxidant results of various eucommia ulmoides extracts
Anti-inflammatory activity of eucommia ulmoides extract
Experimental samples: the fermented extract of eucommia ulmoides of each example and the extract of eucommia ulmoides of comparative example were prepared.
Experimental method, Raw264.7 cells are treated with 5 × 104one/mL of the cells were inoculated in a 24-well plate at 37 ℃ with 5% CO2Culturing for 24h under the condition, adding the extract to be detected, simultaneously taking LPS (sigma, 025M4040V, 1 ten thousand units/mL) as a model control, continuously culturing for 24h, and detecting inflammatory factors by using an ELISA kit.
The experimental conclusion is that the secretion amounts of TNF- α and IL-1 β of mouse macrophage are increased under the stimulation of LPS, and after the mouse macrophage is treated by eucommia bark extracting solution, compared with a model group, the secretion amounts of TNF- α and IL-1 β are reduced, but the reduction degrees are different, compared with comparative example 1, comparative example 2, comparative example 3 and comparative example 4, the TNF- α inhibition rate and the IL-1 β inhibition rate are obviously improved, which shows that the anti-inflammatory ability of synergistic fermentation is more prominent than that of a single strain and only enzymolysis eucommia bark, and the synergistic effect of synergistic fermentation of three strains is shown.
TABLE 2 influence of eucommia ulmoides Oliver extract on TNF- α secretion
| Name (R) | Concentration (pg/mL) | TNF- α inhibition ratio (%) |
| Normal group | 80.42 | - |
| Model set | 310.15 | - |
| Example 1 | 197.34 | 49.11 |
| Example 2 | 212.45 | 42.53 |
| Example 3 | 258.13 | 22.64 |
| Example 4 | 228.04 | 35.74 |
| Example 5 | 245.29 | 28.23 |
| Example 6 | 265.13 | 19.60 |
| Comparative example 1 | 305.18 | 2.16 |
| Comparative example 2 | 292.67 | 7.61 |
| Comparative example 3 | 290.35 | 8.62 |
| Comparative example 4 | 300.89 | 4.03 |
| Comparative example 5 | 285.32 | 10.81 |
TABLE 3 influence of eucommia ulmoides extract on IL-1 β secretion
While embodiments of the present application have been described above, the present application is not limited to the specific embodiments and applications described above, which are intended to be illustrative, instructive, and not limiting. Those skilled in the art, having the benefit of this disclosure, may effect numerous modifications thereto and changes may be made without departing from the scope of the invention as defined by the appended claims.
Claims (10)
1. A preparation method of eucommia ulmoides fermentation extracting solution comprises the following steps:
a step of preparing a composite strain, in which Acetobacter pasteurianus (Acetobacter pasteurium), Lactobacillus plantarum (Lactobacillus plantarum) and Saccharomyces cerevisiae (Saccharomyces cerevisiae) are pre-cultured, respectively, to obtain a composite strain;
the method comprises the steps of performing enzymolysis on eucommia ulmoides powder to prepare a fermentation substrate, wherein in the step, eucommia ulmoides leaves are cleaned, dried and crushed into powder, water is added to the powder to be uniformly mixed, then protease, cellulase and pectinase are added to the powder for enzymolysis, so that an enzymolysis liquid of the eucommia ulmoides is obtained, and the fermentation substrate is prepared on the basis of the enzymolysis liquid;
a step of fermentation, in which the composite strain is inoculated into a fermentation substrate containing the enzymolyzed eucommia ulmoides enzymolysis liquid in a total inoculation amount of 5-10% of the volume of the fermentation substrate, and fermentation is carried out under a low dissolved oxygen condition to obtain fermentation liquid;
and a purification step of centrifuging and filtering the fermentation liquid to obtain eucommia ulmoides fermentation extract.
2. The method of claim 1, wherein, in the step of preparing the composite bacterial strain, the pre-culture solution of the three bacterial strains are obtained by pre-culturing the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae respectively, and in order to ferment by using the composite bacterial strain, the volume ratio of the pre-culture solution of the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae inoculated in the subsequent fermentation process is: 1: 1-6: 0.6 to 4;
wherein, in the pre-culture solution, the bacteria concentration of the acetobacter pasteurianus, the lactobacillus plantarum and the saccharomyces cerevisiae meets the following requirements:
the bacteria concentration in the pre-culture solution of Acetobacter pasteurianus is 1 × 107More than cfu/mL, and the like,
the concentration of the Lactobacillus plantarum preculture solution was 8 × 108More than cfu/mL, and the like,
the concentration of the bacteria in the pre-culture solution of the saccharomyces cerevisiae is 3 × 108cfu/mL or more.
3. The method as claimed in claim 1, wherein in the step of enzymatically hydrolyzing the eucommia ulmoides oliv powder to prepare the fermentation substrate, the mass fraction of the eucommia ulmoides oliv leaves is 5% to 15%, the amount of the protease added is 0.1% to 0.5% of the weight of the selected eucommia ulmoides oliv leaves, the amount of the cellulase added is 0.2% to 1.0% of the weight of the selected eucommia ulmoides oliv leaves, the amount of the pectinase added is 0.1% to 1.0% of the weight of the selected eucommia ulmoides oliv leaves, and the specific activities of the enzymes are not less than 3000U/g.
4. The method as set forth in claim 1, wherein in the step of enzymatically hydrolyzing the eucommia ulmoides powder to prepare the fermentation substrate, the enzymatic hydrolysis reaction conditions are: the pH value is 3.5-6.5, the temperature is 45-60 ℃, the enzymolysis time is 1-3.5 h, and when the enzymolysis process is determined to be finished, the mixture is boiled for 3-5 min to inactivate the enzyme.
5. The method of claim 1, wherein the fermentation substrate comprises, in addition to the eucommia ulmoides enzymolysis solution: a nitrogen source, a carbon source, inorganic salt and the balance of drinking water, wherein the nitrogen source is selected from any one of yeast powder, tryptone, soy peptone or beef extract; the carbon source is selected from any one of sucrose, glucose, lactose or galactose; the inorganic salt is selected from NaCl and K2HPO4。
6. The method according to claim 5, wherein the nitrogen source is 1 to 5 parts, the carbon source is 1 to 5 parts, NaCl is 0.1 to 0.5 part, and K is added to 100 parts by weight of the fermentation substrate2HPO40.1 to 0.5 portion.
7. The method according to claim 1, wherein in the step of performing fermentation, the fermentation temperature is 30 ℃ to 35 ℃, acetobacter pasteurianus, lactobacillus plantarum and saccharomyces cerevisiae are sequentially or simultaneously inoculated, preferably simultaneously inoculated, the pH of the fermentation substrate is adjusted to 6.0 to 7.0, the fermentation time is 36h to 48h, the dissolved oxygen during the culture is controlled to 10% to 40%, and the end of fermentation is judged if no residual carbon source is detected.
8. The method of claim 1, wherein the conditions of centrifugation during purification are set as: centrifuging at 9000rpm for at least 20min to remove thallus; preferably, in the purification process, the filtration conditions are set as follows: adding 1% active carbon, adsorbing at 55 deg.C for 1 hr, and filtering with 1.2 μm fine filter plate and 0.22 μm fine filter core.
9. A fermented extract of eucommia ulmoides prepared by the method according to any one of claims 1 to 8.
10. Use of the fermented extract of eucommia ulmoides as claimed in claim 9 for preparing cosmetics.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010143510.XA CN111317694B (en) | 2020-03-04 | 2020-03-04 | Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010143510.XA CN111317694B (en) | 2020-03-04 | 2020-03-04 | Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111317694A true CN111317694A (en) | 2020-06-23 |
| CN111317694B CN111317694B (en) | 2022-07-19 |
Family
ID=71173135
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010143510.XA Active CN111317694B (en) | 2020-03-04 | 2020-03-04 | Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111317694B (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112870130A (en) * | 2021-03-30 | 2021-06-01 | 中国科学院合肥物质科学研究院 | Preparation method of eucommia pollen fermentation liquor and application of eucommia pollen fermentation liquor in cosmetics |
| CN113384510A (en) * | 2021-06-25 | 2021-09-14 | 科丝美诗(中国)化妆品有限公司 | Fermented composition with anti-photoaging, relieving and cooling effects and preparation method and application thereof |
| CN113564212A (en) * | 2021-07-26 | 2021-10-29 | 浙江树人学院（浙江树人大学） | Method for extracting eucommia ulmoides leaf polysaccharide by using microbial fermentation method |
| CN113768143A (en) * | 2021-05-26 | 2021-12-10 | 商洛学院 | Eucommia ulmoides peptide total nutrient powder and preparation method thereof |
| CN114246192A (en) * | 2021-12-30 | 2022-03-29 | 烟台凯多海洋生物研究院有限公司 | Preparation method and application of affine cudweed botanical insecticide |
| CN114561437A (en) * | 2022-03-24 | 2022-05-31 | 中国科学院广州能源研究所 | Method for preparing functional bacterial cellulose by in-situ fermentation of eucommia ulmoides hydrolysate |
| ES2919234A1 (en) * | 2021-01-21 | 2022-07-22 | Cosmetica Cosbar S L | Vegetable hydrolyzed for use in cosmetics (Machine-translation by Google Translate, not legally binding) |
| CN115006295A (en) * | 2022-07-06 | 2022-09-06 | 上海云未生物科技有限公司 | Anti-aging composition, preparation method and application thereof in preparation of anti-aging cosmetics |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170072162A (en) * | 2015-12-16 | 2017-06-26 | 계명대학교 산학협력단 | Mixed Fermented Material of Extraction of Oriental Medicine Material With Higher γ-PGA and GABA for Enhancing Immune Activity or Inhibiting Allergy and Method for Manufacturing the Same |
| CN107549563A (en) * | 2017-09-29 | 2018-01-09 | 贵州五新农业科技有限责任公司 | Bark of eucommia beverage and preparation method thereof |
| CN107927497A (en) * | 2017-11-24 | 2018-04-20 | 山东贝隆杜仲生物工程有限公司 | A kind of production method of bark of eucommia fermented beverage |
| CN109498744A (en) * | 2018-08-24 | 2019-03-22 | 四川长重宝生物工程有限责任公司 | A kind of preparation method that composite bacteria fermentation compound eucommia bark leaf is scattered |
| KR102007144B1 (en) * | 2019-04-26 | 2019-08-02 | 에스케이바이오랜드 주식회사 | A novel strain of Lactobacillus plantarum SKB1234 having an improving effect on arthritis and a method for producing a mixture of the fermented Achyranthes japonica Nakai extract using the Lactobacillus plantarum SKB1234, Angelica gigas Nakai extract and Eucommia ulmoides Oliver extract |
-
2020
- 2020-03-04 CN CN202010143510.XA patent/CN111317694B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170072162A (en) * | 2015-12-16 | 2017-06-26 | 계명대학교 산학협력단 | Mixed Fermented Material of Extraction of Oriental Medicine Material With Higher γ-PGA and GABA for Enhancing Immune Activity or Inhibiting Allergy and Method for Manufacturing the Same |
| CN107549563A (en) * | 2017-09-29 | 2018-01-09 | 贵州五新农业科技有限责任公司 | Bark of eucommia beverage and preparation method thereof |
| CN107927497A (en) * | 2017-11-24 | 2018-04-20 | 山东贝隆杜仲生物工程有限公司 | A kind of production method of bark of eucommia fermented beverage |
| CN109498744A (en) * | 2018-08-24 | 2019-03-22 | 四川长重宝生物工程有限责任公司 | A kind of preparation method that composite bacteria fermentation compound eucommia bark leaf is scattered |
| KR102007144B1 (en) * | 2019-04-26 | 2019-08-02 | 에스케이바이오랜드 주식회사 | A novel strain of Lactobacillus plantarum SKB1234 having an improving effect on arthritis and a method for producing a mixture of the fermented Achyranthes japonica Nakai extract using the Lactobacillus plantarum SKB1234, Angelica gigas Nakai extract and Eucommia ulmoides Oliver extract |
Non-Patent Citations (4)
| Title |
|---|
| 冷桂华等: "杜仲叶发酵液的工艺研究", 《中国酿造》 * |
| 张学梅: "利用杜仲叶中主要有效成分研制杜仲粉", 《中国优秀博硕士学位论文全文数据库(硕士)工程科技Ⅰ辑》 * |
| 李强等: "《新编常用中药有效成分手册》", 31 January 2008, 中国协和医科大学出版社 * |
| 汪勇: "《粮油副产物加工学》", 31 January 2019, 暨南大学出版社 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2919234A1 (en) * | 2021-01-21 | 2022-07-22 | Cosmetica Cosbar S L | Vegetable hydrolyzed for use in cosmetics (Machine-translation by Google Translate, not legally binding) |
| CN112870130A (en) * | 2021-03-30 | 2021-06-01 | 中国科学院合肥物质科学研究院 | Preparation method of eucommia pollen fermentation liquor and application of eucommia pollen fermentation liquor in cosmetics |
| CN113768143A (en) * | 2021-05-26 | 2021-12-10 | 商洛学院 | Eucommia ulmoides peptide total nutrient powder and preparation method thereof |
| CN113384510A (en) * | 2021-06-25 | 2021-09-14 | 科丝美诗(中国)化妆品有限公司 | Fermented composition with anti-photoaging, relieving and cooling effects and preparation method and application thereof |
| CN113384510B (en) * | 2021-06-25 | 2023-01-17 | 科丝美诗(中国)化妆品有限公司 | Fermented composition with anti-photoaging, relieving and cooling effects and preparation method and application thereof |
| CN113564212A (en) * | 2021-07-26 | 2021-10-29 | 浙江树人学院（浙江树人大学） | Method for extracting eucommia ulmoides leaf polysaccharide by using microbial fermentation method |
| CN113564212B (en) * | 2021-07-26 | 2024-01-16 | 浙江树人学院（浙江树人大学） | Method for extracting eucommia ulmoides leaf polysaccharide by utilizing microbial fermentation method |
| CN114246192A (en) * | 2021-12-30 | 2022-03-29 | 烟台凯多海洋生物研究院有限公司 | Preparation method and application of affine cudweed botanical insecticide |
| CN114561437A (en) * | 2022-03-24 | 2022-05-31 | 中国科学院广州能源研究所 | Method for preparing functional bacterial cellulose by in-situ fermentation of eucommia ulmoides hydrolysate |
| CN115006295A (en) * | 2022-07-06 | 2022-09-06 | 上海云未生物科技有限公司 | Anti-aging composition, preparation method and application thereof in preparation of anti-aging cosmetics |
| CN115006295B (en) * | 2022-07-06 | 2023-06-23 | 上海云未生物科技有限公司 | Anti-aging composition, preparation method and application thereof in preparation of anti-aging cosmetics |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111317694B (en) | 2022-07-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111317694B (en) | Eucommia ulmoides fermentation extracting solution, preparation method thereof and application thereof in cosmetics | |
| CN109717340A (en) | A kind of fermentation preparation of the two-step Cordyceps militaris ferment of combination complex enzyme hydrolysis | |
| CN104013657A (en) | Post-fermentation extracting method of extracting saponin from American ginseng medicinal material | |
| CN112999127B (en) | Gentiana scabra bunge compound enzyme and preparation method and application thereof | |
| CN109593664B (en) | Candida ethanolica and application thereof | |
| CN113966832A (en) | Dendrobium nobile fermentation product and preparation method and application thereof | |
| CN116064685B (en) | Preparation process and application of eurotium cristatum fermented edible traditional Chinese medicine | |
| CN115572738A (en) | Dendrobium officinale fermentation liquor with anti-inflammatory, relieving and repairing effects as well as preparation method and application thereof | |
| CN114304335A (en) | Method for enriching active ingredients of dendrobium leaves through fermentation and application of method | |
| CN104357332B (en) | Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid | |
| CN114107403B (en) | Method for co-producing ellagic acid and biological feed by fermenting pericarpium Granati with microbial community | |
| CN112608949B (en) | Preparation method and application of pseudo-ginseng flower extract | |
| CN111961698A (en) | Preparation method and application of passion fruit peel polysaccharide degraded by lactobacillus brevis enzyme production | |
| CN111743837A (en) | Fermented traditional Chinese medicine mask with whitening effect and preparation method thereof | |
| KR102011133B1 (en) | Lactobacillus plantarum MBE/L2990 strain having excellent alpha-rhamnosidase activity and bioconversion activity from ginsenoside Re and Rb1 to ginsenoside Rg1 and Rg5 | |
| CN111449239A (en) | Functional food additive of ganoderma lucidum fermented sea buckthorn seed meal and preparation method thereof | |
| CN111494255B (en) | Composition containing ginseng cordyceps sinensis fermentation extract and application of composition in cosmetics | |
| CN109609395B (en) | Pichia corylifolia and application thereof | |
| CN112877157A (en) | Preparation method of radix puerariae wine | |
| CN111149961A (en) | Grape health-care beverage and preparation method thereof | |
| CN114875104B (en) | Mulberry leaf ferment stock solution and preparation method and application thereof | |
| AU2020102037A4 (en) | A method of efficiently increasing the alpha-glucosidase inhibitor content in fresh mulberry leaves by the solid-state fermentation | |
| CN114250252B (en) | Method for preparing multifunctional glycerin by fermenting raisins | |
| CN116115675B (en) | Method for extracting loquat leaf total flavonoids by microbial enzymolysis | |
| CN111304122B (en) | Process for modifying hemlock parsley by using microorganisms and application of process |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230602 Address after: Tianchen Avenue, Ji'nan hi tech Development Zone of Shandong Province, No. 678 250101 Patentee after: BLOOMAGE BIOTECH Co.,Ltd. Address before: No. 678, Tianchen street, high tech Zone, Jinan City, Shandong Province Patentee before: BLOOMAGE BIOTECH Co.,Ltd. Patentee before: SHANDONG BLOOMAGE HYINC BIOPHARM Corp.,Ltd. |
|
| TR01 | Transfer of patent right |