CN116270826A - Method for improving activity of rose in reducing blood sugar and blood fat and application - Google Patents
Method for improving activity of rose in reducing blood sugar and blood fat and application Download PDFInfo
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- 241000220317 Rosa Species 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 38
- 239000008280 blood Substances 0.000 title abstract description 31
- 210000004369 blood Anatomy 0.000 title abstract description 31
- 230000000694 effects Effects 0.000 title abstract description 15
- 238000000855 fermentation Methods 0.000 claims abstract description 37
- 230000004151 fermentation Effects 0.000 claims abstract description 37
- 239000000284 extract Substances 0.000 claims abstract description 23
- 239000008132 rose water Substances 0.000 claims abstract description 22
- 241000235342 Saccharomycetes Species 0.000 claims abstract description 17
- 241000894006 Bacteria Species 0.000 claims abstract description 15
- 241000186660 Lactobacillus Species 0.000 claims abstract description 14
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 16
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 16
- 239000000047 product Substances 0.000 claims description 16
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 15
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 15
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 15
- 238000002360 preparation method Methods 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 9
- 239000004310 lactic acid Substances 0.000 claims description 8
- 235000014655 lactic acid Nutrition 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 6
- 230000000055 hyoplipidemic effect Effects 0.000 claims description 6
- 235000004789 Rosa xanthina Nutrition 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
- 241000109329 Rosa xanthina Species 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 230000002218 hypoglycaemic effect Effects 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 241000894007 species Species 0.000 claims 1
- 230000000975 bioactive effect Effects 0.000 abstract description 11
- 239000000126 substance Substances 0.000 abstract description 8
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- 239000004480 active ingredient Substances 0.000 abstract description 4
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- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 6
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 6
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 6
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 6
- 235000005493 rutin Nutrition 0.000 description 6
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- 230000004913 activation Effects 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 238000004321 preservation Methods 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
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- 102000019280 Pancreatic lipases Human genes 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 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 4
- 238000012258 culturing Methods 0.000 description 4
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- 229940074391 gallic acid Drugs 0.000 description 4
- 235000004515 gallic acid Nutrition 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 229940116369 pancreatic lipase Drugs 0.000 description 4
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 4
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 4
- 238000003809 water extraction Methods 0.000 description 4
- LWFUFLREGJMOIZ-UHFFFAOYSA-N 3,5-dinitrosalicylic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O LWFUFLREGJMOIZ-UHFFFAOYSA-N 0.000 description 3
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- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 description 2
- OOSZCNKVJAVHJI-UHFFFAOYSA-N 1-[(4-fluorophenyl)methyl]piperazine Chemical compound C1=CC(F)=CC=C1CN1CCNCC1 OOSZCNKVJAVHJI-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 235000011449 Rosa Nutrition 0.000 description 2
- 239000003392 amylase inhibitor Substances 0.000 description 2
- 229930002877 anthocyanin Natural products 0.000 description 2
- 235000010208 anthocyanin Nutrition 0.000 description 2
- 239000004410 anthocyanin Substances 0.000 description 2
- 150000004636 anthocyanins Chemical class 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- DGLRDKLJZLEJCY-UHFFFAOYSA-L disodium hydrogenphosphate dodecahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.O.O.[Na+].[Na+].OP([O-])([O-])=O DGLRDKLJZLEJCY-UHFFFAOYSA-L 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
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- 239000012488 sample solution Substances 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 229940074545 sodium dihydrogen phosphate dihydrate Drugs 0.000 description 2
- 235000010288 sodium nitrite Nutrition 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 238000009966 trimming Methods 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 1
- 241000099774 Cuscuta salina Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 244000181025 Rosa gallica Species 0.000 description 1
- 235000000533 Rosa gallica Nutrition 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 150000001413 amino acids Chemical class 0.000 description 1
- 239000006286 aqueous extract Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 235000010633 broth Nutrition 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- HVQAJTFOCKOKIN-UHFFFAOYSA-N flavonol Natural products O1C2=CC=CC=C2C(=O)C(O)=C1C1=CC=CC=C1 HVQAJTFOCKOKIN-UHFFFAOYSA-N 0.000 description 1
- 150000007946 flavonol Chemical class 0.000 description 1
- 235000011957 flavonols Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 235000019626 lipase activity Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000874 microwave-assisted extraction Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 description 1
- 229940074439 potassium sodium tartrate Drugs 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229940079827 sodium hydrogen sulfite Drugs 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 235000011006 sodium potassium tartrate Nutrition 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/738—Rosa (rose)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Diabetes (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Epidemiology (AREA)
- Medical Informatics (AREA)
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Abstract
The invention provides a method for improving the activity of rose in reducing blood sugar and blood fat, which comprises the following steps: inoculating mixed strains of lactobacillus and saccharomycetes into the rose water extract for fermentation to obtain rose fermentation product. The invention adopts the mixed bacteria of lactobacillus and saccharomycetes to ferment the rose water extract, and various secondary metabolites including enzymes produced in the fermentation process can hydrolyze bioactive components into substances with smaller molecular weight so as to enhance the bioactivity and promote the biological accessibility of bioactive components. The obtained rose fermented product has high content of active ingredients and obvious effects of reducing blood sugar and blood fat. In addition, the method provided by the invention has the advantages of economic cost and higher safety.
Description
Technical Field
The invention belongs to the technical field of natural product extraction and biological medicine, and particularly relates to a method for improving the activity of rose in reducing blood sugar and blood fat and application thereof.
Background
Roses (Rosa rugasa Thunb), also called red rose, loitering rose and the like, are rose-ornamental fallen leaf shrubs of Rosa genus of Rosaceae, have a cultivation history of over 2000 years in China, and are planted in Shandong, gansu, yunnan and other places. The flower contains various nutritional components such as fat, amino acids, vitamins, etc., and various bioactive components such as flavonoid, anthocyanin, polyphenols, volatile oil, etc. The rose is one of the traditional medicinal materials in China, and the description of the Chinese pharmacopoeia is recorded: the flower shape Wen Weigan can promote qi circulation, relieve depression, relieve pain and promote blood circulation; in addition, many pharmacological researches show that the compound has various physiological activity functions of resisting oxidation, aging, tumor, inflammation, bacteria and blood sugar, and the like.
The rose has rich content of bioactive substances such as polyphenols, anthocyanin, flavonol and the like, and a great deal of literature reports that the components have potential pharmacological effects of resisting oxidation, reducing blood sugar, reducing blood fat, resisting inflammation and the like. The method for extracting the bioactive components mainly comprises physical and chemical methods such as hot water extraction, organic solvent extraction, supercritical extraction, microwave extraction and the like, but the rose obtained by the method has low content of active ingredients and limited efficacy of reducing blood sugar and blood fat.
Disclosure of Invention
In view of the above, the technical problem to be solved by the invention is to provide a method for improving the activity of reducing blood sugar and blood fat of roses and application thereof.
The invention provides a method for improving the activity of rose in reducing blood sugar and blood fat, which comprises the following steps:
inoculating mixed strains of lactobacillus and saccharomycetes into the rose water extract for fermentation to obtain rose fermentation product.
Preferably, the rose water extract is prepared according to the following method:
mixing the dry powder of the rose with water, heating and extracting, centrifuging the extracting solution, and taking the supernatant to obtain the rose water extract.
Preferably, the mass ratio of the rose dry powder to the water is 1: (20-60);
the temperature of the heating extraction is 90-95 ℃ and the time is 1-1.5 h.
Preferably, the lactic acid bacteria are selected from lactobacillus plantarum.
Preferably, the lactobacillus plantarum is selected from the group consisting of the lactobacillus plantarum with a preservation number of CCTCC No: m20191045 Lactobacillus plantarum Picp-2.
Preferably, the saccharomycete is selected from saccharomycete with a preservation number of CCTCC KY2008671 (Y671) in China center for type culture collection; one or more of yeasts with deposit number ACCC20163 (A163) and yeasts with deposit number ACCC 21190 (A190) of China center for type culture Collection of agricultural microorganisms.
Preferably, the mass ratio of the lactobacillus to the saccharomycete is (2-5): (0.5-1.5);
the inoculation amount of the mixed strain in the rose water extract is 2.5% -6.5%.
Preferably, the temperature of the fermentation is 20-30 ℃ and the time is 7-14 days.
The invention also provides the rose fermentation product prepared by the method.
The invention also provides application of the rose fermentation product in preparation of products with the effects of reducing blood sugar and/or blood fat.
Compared with the prior art, the invention provides a method for improving the activity of rose in reducing blood sugar and blood fat, which comprises the following steps: inoculating mixed strains of lactobacillus and saccharomycetes into the rose water extract for fermentation to obtain rose fermentation product. The invention adopts the mixed bacteria of lactobacillus and saccharomycetes to ferment the rose water extract, and various secondary metabolites including enzymes produced in the fermentation process can hydrolyze bioactive components into substances with smaller molecular weight so as to enhance the bioactivity and promote the biological accessibility of bioactive components. The obtained rose fermented product has high content of active ingredients and obvious effects of reducing blood sugar and blood fat. In addition, the method provided by the invention has the advantages of economic cost and higher safety.
Detailed Description
The invention provides a method for improving the activity of rose in reducing blood sugar and blood fat, which comprises the following steps:
inoculating mixed strains of lactobacillus and saccharomycetes into the rose water extract for fermentation to obtain rose fermentation product.
The invention firstly prepares a rose water extract, and specifically, the rose water extract is prepared according to the following method:
mixing the dry powder of the rose with water, heating and extracting, centrifuging the extracting solution, and taking the supernatant to obtain the rose water extract.
Wherein the rose is selected from the group consisting of rose with golden edge.
The mass ratio of the rose dry powder to the water is 1: (20-60), preferably 1:20, 1:30, 1:40, 1:50, 1:60, or 1: any value between (20) and (60);
the temperature of the heating extraction is 90-95 ℃, preferably 90, 91, 92, 93, 94, 95, or any value between 90-95 ℃ for 1-1.5 h.
The method of centrifugation is not particularly limited, and may be any method known to those skilled in the art. In the present invention, it is preferable to perform centrifugation according to the process parameters of 9000r/min for 15 min.
And after centrifugation, removing supernatant fluid to obtain the rose water extract.
Then inoculating mixed strains of lactobacillus and saccharomycetes into the rose water extract for fermentation.
Wherein the lactic acid bacteria and the yeast need to be activated before inoculation, the method of the activation is not particularly limited, and the activation method known to those skilled in the art can be used.
Preferably, the activation is performed as follows:
yeast: inoculating the strain into YPD culture medium, culturing at 30deg.C for 36-48 hr, and continuously passaging for 3 times to fully activate the strain until the number of viable bacteria is 1.0X10 7 CFU/mL or more.
Lactic acid bacteria: inoculating strain into MRS culture medium, culturing at 35deg.C for 24 hr, and continuously passaging for 3 times to fully activate strain until viable count is 1.0X10 8 CFU/mL or more.
In the invention, the lactobacillus is selected from lactobacillus plantarum, and the lactobacillus plantarum is selected from the group consisting of the lactobacillus plantarum with the preservation number of CCTCC No: m20191045 Lactobacillus plantarum Picp-2.
The saccharomycete is selected from saccharomycete with a preservation number of CCTCC KY2008671 (Y671) of China center for type culture collection; one or more of yeasts with deposit number ACCC20163 (A163) and yeasts with deposit number ACCC 21190 (A190) of China center for type culture Collection of agricultural microorganisms.
The mass ratio of the lactobacillus to the saccharomycete is (2-5): (0.5 to 1.5), preferably 2.5:1, 2:0.5, 2:1.5, 5:0.5, 5:1.5, or (2 to 5): (0.5 to 1.5);
the inoculation amount of the mixed bacteria in the rose water extract is 2.5% -6.5%, preferably 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5% or any value between 2.5% -6.5%.
In the present invention, the mixed strain of lactic acid bacteria and yeast is inoculated into the aqueous extract of rose flowers to ferment at 20 to 30 ℃, preferably at 20, 25, 30, or at any value between 20 and 30 ℃ for 7 to 14 days, preferably at any value between 7, 8, 10, 12, 14, or 7 to 14 days.
And after fermentation is completed, centrifuging the fermentation liquor, and taking supernatant of the centrifuged fermentation liquor to obtain the rose fermentation liquor.
The invention also provides the rose fermentation product prepared by the method.
The invention also provides application of the rose fermentation product in preparation of products with the effects of reducing blood sugar and/or blood fat.
The invention adopts the mixed bacteria of lactobacillus and saccharomycetes to ferment the rose water extract, and various secondary metabolites including enzymes produced in the fermentation process can hydrolyze bioactive components into substances with smaller molecular weight so as to enhance the bioactivity and promote the biological accessibility of bioactive components. The obtained rose fermented product has high content of active ingredients and obvious effects of reducing blood sugar and blood fat. In addition, the method provided by the invention has the advantages of economic cost and higher safety.
For further understanding of the present invention, the method and application of the present invention for improving the hypoglycemic and hypolipidemic activities of roses are described below with reference to examples, and the scope of the present invention is not limited by the following examples.
The lactobacillus plantarum is selected from the CCTCC No: m20191045 Lactobacillus plantarum Picp-2.
The saccharomycete is selected from one or more of yeasts Y671, A163 and A190.
Wherein the microzyme Y671 is selected from China Center for Type Culture Collection (CCTCC) KY2008671;
a163 is selected from China center for type culture Collection (ACCC) 20163;
a190 is selected from China center for type culture Collection, accession number ACCC 21190.
The flos Rosae Rugosae is selected from flos Rosae Rugosae with golden thread
Example 1
(1) Rose water extraction
Water: the rose dry powder=60:1 mass ratio, extracting for 3 hours at 95 ℃, centrifuging for 15 minutes at 9000r/min, and taking the supernatant as the water extract.
(2) Preparation of water extracted rose fermented product
Fermentation strain selection and activation
A) Fermentation strain selection
Lactic acid bacteria: lactobacillus plantarum (Lactobacillus plantarum Picp-2 with preservation number of CCTCC No. M20191045)
Yeast: y671, A163, A190 (Y671 number CCTCC KY2008671; A163 number ACCC 201663; A190 number ACCC 21190).
B) Fermentation strain activation
Yeast: inoculating strain into YPD culture medium, culturing at 30deg.C for 36 hr, and continuously passaging for 3 times to fully activate strain until viable count is 1.0X10 7 CFU/mL or more.
Lactic acid bacteria: inoculating strain into MRS culture medium, culturing at 35deg.C for 24 hr, and continuously passaging for 3 times to fully activate strain until viable count is 1.0X10 8 CFU/mL or more.
C) Single-bacteria fermentation of rose after water extraction
The rose water extract is packaged in 250mL blue-cap reagent bottles (200 mL/bottle), sterilized (115 ℃ C., 30 min) and prepared for fermentation.
Inoculating activated lactobacillus and yeast strain respectively at an inoculation amount of 5%, mixing, placing in an incubator at 37deg.C, fermenting for 7 days, and centrifuging at 6000r/min for 10min to obtain flos Rosae Rugosae ferment for index detection.
D) Complex bacteria fermentation after water extraction of rose
The rose water extract is packaged in 250mL blue-cap reagent bottles (200 mL/bottle), sterilized (115 ℃ C., 30 min) and prepared for fermentation.
Firstly, inoculating activated lactobacillus strains and 1% of saccharomycetes according to the proportion of 2.5% of inoculum size, uniformly mixing, placing in an incubator at 28 ℃, fermenting for 7d, and centrifuging for 15min at 9000r/min to obtain a rose fermentation product for index detection.
Example 2
1. Determination of bioactive substances
1.1 Total polyphenols
(1) Reagent(s)
Gallic acid solution
20% sodium carbonate solution
Fu Lin Fen solution
(2) Experimental procedure
Drawing a standard curve:
0.1ml of gallic acid with concentration of 0, 0.01, 0.02, 0.04, 0.06 and 0.08mg/ml is taken in a 2ml EP tube, 0.05ml of Fu Lin Fen is added, and after mixing, the mixture reacts for 3 to 8 minutes at room temperature, then 0.15ml of 20 percent sodium carbonate solution is added, and distilled water is added for constant volumeTo 1ml, the absorbance was measured at 765nm luminosity after 60min of reaction. The total polyphenol content is expressed by the concentration of gallic acid (mg/ml), the total polyphenol content is taken as an abscissa, the absorbance value is taken as an ordinate, and an XY curve graph of a standard curve (scatter diagram) of the total polyphenol solution is drawn, and a correlation coefficient R is obtained 2 As close to 0.999 as possible.
Determination of total polyphenol content of samples:
taking 1ml of a sample solution to be tested centrifuged after fermentation, reacting according to a gallic acid standard curve drawing method, measuring absorbance A at 765nm wavelength, and determining the absorbance A according to a regression equation (y=0.0063x+0.0099, R 2 =0.9982) to calculate the total polyphenol content (mg/ml) in the test solution.
(3) Experimental results
See Table 1
1.2 Total Flavonoids
(1) Sample and reagent preparation
Reagent:
rutin standard substance not less than 98%, ethanol with purity of 75%, sodium nitrite solution with purity of 5%, aluminum nitrate solution with purity of 10.00% and sodium hydroxide solution with purity of 4.00%
(2) Experimental procedure
Drawing and process of rutin standard curve
Preparing standard solution by adding 5mg rutin standard into 25ml volumetric flask, adding 70% ethanol to scale, shaking to obtain 0.2mg/ml rutin standard solution, respectively weighing 0.00, 1.00, 2.00, 3.00, 4.00, 5.00, 6.00ml rutin standard solution, respectively placing into 25ml volumetric flask, adding 6.00ml water, adding 5% sodium nitrite solution 1.00ml shaking, standing for 6min, adding 10.00ml aluminum nitrate solution 1.00ml shaking, standing for 6min, adding 4.00% sodium hydroxide solution 10.00ml, respectively adding water to scale, standing for 15min, and measuring absorbance at 510nm wavelength with maximum absorption wavelength of 510nm by ultraviolet spectrophotometry Ji Quanbo long 200-800 nm scanning, and making standard curve (y= 1.2954x-0.0027, R) with concentration C as horizontal coordinate and absorbance A as vertical coordinate 2 ＝0.9998)
Determination of total flavone content of the sample:
taking 1ml of a sample solution to be detected according to a rutin standard curveThe absorbance A was measured at a wavelength of 510nm after the drawing method was left standing for 15 minutes, and according to the regression equation (y= 1.2954x-0.0027, R 2 =0.9998) to calculate the total flavone content (mg/ml) in the test solution.
(3) Experimental results
See Table 1
TABLE 1 determination of bioactive substances of rose fermentation broths
In Table 1, the fruit and vegetable fermenters are from Angel Yeast Co.
The dosage ratio of the Picp-2 plus fruit and vegetable ferment is 1:1
As is clear from the data in Table 1, the total polyphenol content in the sample fermented by the Y671 strain was the highest, 2.680mg/ml, and the total flavone content in the sample fermented by the A163 strain was the highest, 0.263mg/ml. In the mixed fermentation, the content of total polyphenol in the Picp-2+ fruit and vegetable starter is 2.096mg/ml, and the content of total flavone in a sample fermented by the Picp-2+ A163 strain is 0.219mg/ml.
2.1 alpha-Amylase Activity inhibition
(1) Reagent preparation
25 mmph=6.9 PBS: 4.477g of disodium hydrogen phosphate dodecahydrate and 1.95g of sodium dihydrogen phosphate dihydrate are respectively dissolved in 500ml of distilled water, and after uniform mixing, ph is adjusted to 6.9;
0.1U/mL of alpha-amylase (alpha-amylase) solution: 13.51mg of alpha-amylase was weighed into 50ml of PBS;
1% soluble starch solution (soluble starch): weighing 1g of soluble starch, dissolving in 100ml of boiling water, and boiling for 2-3min;
DNS preparation: (Ghose method)
And (2) solution A: 6.9g of crystalline phenol was dissolved in 15.2ml of 10% NaOH solution, distilled water was diluted to 69ml, and 6.9g of sodium hydrogensulfite was added to the solution;
b, liquid: 255g of potassium sodium tartrate solution are dissolved in 300ml of 10% NaOH solution, and 880ml of 1% 3, 5-dinitrosalicylic acid solution are added;
the orange reagent can be obtained by mixing the first and second solutions, and can be stored in a brown bottle for one week for use, and can be stored in the brown bottle for one year.
(2) Detailed description of the invention
Mu.l of the sample was mixed with 50. Mu.l of 0.1U/mL of the alpha-amylase solution and 400. Mu.l of the substrate 1% soluble starch uniformly, reacted in a water bath at 37℃for 15 minutes, and then 1.5mL of DNS was added to terminate the reaction in boiling water for 5 minutes. The absorbance was measured at 504nm after the reaction solution was cooled to room temperature, and the α -amylase inhibition rate was calculated according to the following formula.
Alpha-amylase inhibition ratio (%) = [1- (A4-A3)/(A2-A1) ]. Times.100%
Control background group A1: substrate (soluble starch)
Control group A2: enzymes and substrates (soluble starch)
Experimental background group A3: sample+substrate (soluble starch)
Experimental group A4: experimental group
The total system was 2ml and insufficient PBS was added for trimming.
(3) Experimental results
See Table 2
2.2 hypolipidemic Activity
Hypolipidemic regimen
(1) Reagent preparation
Purchased reagents are shown in Table 2
TABLE 2
(2) And (3) preparation of a reagent:
25mM ph=7.4 PBS: 4.477g of disodium hydrogen phosphate dodecahydrate and 1.95g of sodium dihydrogen phosphate dihydrate are respectively dissolved in 500ml of distilled water, and the pH is adjusted to 7.4 after uniform mixing;
pancreatic lipase 5 mg/ml: weighing 50mg of pancreatic lipase, dissolving in 50ml of sterile water, centrifuging at 8000r for 5min, discarding precipitate to obtain supernatant, and placing enzyme in ice chest for use;
preparation of 11.2mol/L p-NPB solution: the 0.11715g p-NPB solution was pipetted into 50ml PBS and placed in an ice box for further use.
(3) Experimental procedure
Mu.l of sample, 500. Mu.l of pancreatic lipase and 500. Mu.l of PBS are mixed and placed in a water bath at 37 ℃ for incubation for 10min, then 500. Mu.l of p-NPB solution is added, the mixture is gently shaken and mixed, placed in a water bath at 37 ℃ for incubation for 20min, absorbance is measured at 405nm wavelength and the value is recorded.
Inhibition ratio (%) = [1- (A4-A3)/(A2-A1) ] ×100%
Control background group A1: substrate(s)
Control group A2: enzymes and substrates
Experimental background group A3: sample+substrate
Experimental group A4: experimental group
The total system was 200. Mu.l and insufficient PBS was added for trimming.
(4) Experimental results
See Table 3
TABLE 3 Table 3
As is clear from Table 3, the samples fermented with the Picp-2+A163 mixture had the highest hypoglycemic activity (inhibition of. Alpha. -amylase activity) and hypolipidemic activity (inhibition of pancreatic lipase activity), 113.43% and 90.30%, respectively.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A method for improving the hypoglycemic and hypolipidemic activities of roses, which is characterized by comprising the following steps:
inoculating mixed strains of lactobacillus and saccharomycetes into the rose water extract for fermentation to obtain rose fermentation product.
2. The method of claim 1, wherein the aqueous rose extract is prepared by the following method:
mixing the dry powder of the rose with water, heating and extracting, centrifuging the extracting solution, and taking the supernatant to obtain the rose water extract.
3. The method of claim 2, wherein the mass ratio of rose dry powder to water is 1: (20-60);
the temperature of the heating extraction is 90-95 ℃ and the time is 1-1.5 h.
4. The method according to claim 1, wherein the lactic acid bacteria are selected from the group consisting of lactobacillus plantarum.
5. The method according to claim 4, wherein the lactobacillus plantarum is selected from the group consisting of the species having a accession number cctccc No: m20191045 Lactobacillus plantarum Picp-2.
6. The method according to claim 1, wherein the yeast is selected from the group consisting of yeasts of the China center for type culture Collection (cctccc KY 2008671) (Y671); one or more of yeasts with deposit number ACCC20163 (A163) and yeasts with deposit number ACCC 21190 (A190) of China center for type culture Collection of agricultural microorganisms.
7. The method according to claim 1, wherein the mass ratio of the lactic acid bacteria to the yeast is (2-5): (0.5-1.5);
the inoculation amount of the mixed strain in the rose water extract is 2.5% -6.5%.
8. The method according to claim 1, wherein the fermentation is carried out at a temperature of 20 to 30 ℃ for a period of 7 to 14 days.
9. A rose fermentation product obtainable by a process according to any one of claims 1 to 8.
10. Use of the rose fermentation according to claim 9 for the preparation of a product having hypoglycemic and/or hypolipidemic effects.
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