CN103263448B - Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application - Google Patents
Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application Download PDFInfo
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- CN103263448B CN103263448B CN201310067394.8A CN201310067394A CN103263448B CN 103263448 B CN103263448 B CN 103263448B CN 201310067394 A CN201310067394 A CN 201310067394A CN 103263448 B CN103263448 B CN 103263448B
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Abstract
The present invention discloses fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and an application method. The fermentation bacteria is aspergillus. The patent accession number is CCTCCM2012515. The method includes firstly crushing Ginkgo biloba L. leaves to 20-60 mesh; mixing the Ginkgo biloba L. leaves with water, wherein the water adding amount is 45%-90%; sterilizing the mixture at 115 DEG C for 15 minutes, filling into a fermenter; inoculating strains as the weight of the Ginkgo biloba L. leaves in the mixture with an inoculating amount of 0.01%-0.02%; performing aerobic fermentation at 25-30 DEG C for 2-11 days; adding a suitable amount of alcohol and water as an ultimate parameter solid-liquid ratio of 1:20 and the ethanol content of 60%, stirring constantly and extracting flavonoid substances; filtering suspension to obtain an extracting solution; and concentrating the extracting solution in vacuum to obtain the finished product, or further drying to a solid. The invention uses Aspergillus niger for fermentation pretreatment of Ginkgo biloba L. leaves to degrade leaf cells, and conventional extraction methods are then used for extraction, so that the extraction rate of flavonoids is improved. Compared with the conventional alcohol solvent extraction, the extraction rate of flavonoids in Ginkgo biloba L. leaves are increased by 20-60%.
Description
Technical field
The invention belongs to ginkgetin extractive technique field, relate in particular to a kind of pretreatment of fermenting and improve fermented bacterium and the application process thereof that ginkgetin extracts.
Background technology
Since last century the eighties, along with improving constantly of China's living standards of the people, people's Dietary Pattern also has a very large change, and the shared ratio of polish food, animal product of absorbing in daily life increases day by day, and the sickness rate of person in middle and old age's cardiovascular disease obviously improves.In China, only hypertension just accounts for the 25%-50% of all death, and the number of dying from every year cardiovascular disease has risen to first or second, and the sickness rate of person in middle and old age's cardiovascular disease is also being the trend of rising in recent years.Therefore, the M & M of prevention and minimizing cardiovascular disease, reasonable, healthy meals are just particularly important.For the seriously ill patient of person in middle and old age's cardiovascular, except actively carrying out medication, dietetic therapy is also a kind of important means of controlling the state of an illness of can yet be regarded as; For the light patient of person in middle and old age cardiovascular diseases, healthy dietetic therapy more should be put in the first place of controlling and alleviating the state of an illness.
Semen Ginkgo (GinkgobilobaL.) is to originate in the ancient Relict Plant of China.Its leaf sweet in the mouth, hardship, puckery; Property is put down, and has the beneficial heart to astringe the lung removing dampness antidiarrheal.Control uncomfortable in chest pained, severe palpitation, phlegm dyspnea cough, dysentery, effect of leucorrhea." product converge with essence will " record Folium Ginkgo " make cake for last dough-making powder, stewing prepared food it, antidiarrheal dysentery "." Treatise on Dietetic Therapy " recorded, and Folium Ginkgo can be used for the disease such as severe palpitation, cough and asthma due to lung deficiency.
Folium Ginkgo is taking flavone as main effective ingredient, content is generally 2.5%-3.8%, containing isorhamnetin, nimbecetin, Kaempferol-Fructus rhamni (Rhamnus davurica Pall.) glycoside, Quercetin, globulariacitrin, quercitrin, ginkegetin, isoginkgetin, ginkgolide A, cachou extract, epicatechin, Chinese infanticide cachou extract composition.Research shows that Folium Ginkgo flavone has protection capillary permeability, coronary artery dilator, recovery arteries elasticity, reduces serum cholesterol, increases coronary artery blood flow, improves cardiovascular and cerebrovascular circulation, removes smooth muscle spasm, the effect of lax bronchus and antibacterial, nourishing brain cell and other organ, and makes in addition blood matter in tremulous pulse, peripheral vessel, blood capillary and cholesterol maintain peculiar effect of normal level.Be used for the treatment of coronary atherosclerotic heart disease, preliminary observation has better effects to angina pectoris, also has certain effect to reducing cholesterol and blood pressure.
Semen Ginkgo extrac product spreads all over market at present, comprises various buccal tablets and oral liquid, and adds the various functional food of extract.The inventive method is simple, science, and production cost is low, and yield of flavone improves significantly, therefore has broad application prospects.
The single alcoholic solution extraction process of the general employing of technology of existing extraction Folium Ginkgo flavone, the efficiency of extraction is low.Main cause is: the granularity limited (20-60 order) that leaf is pulverized, and leaf cell, still in good working condition, has hindered the stripping of intracellular matter mostly, and research shows the meticulous extraction that is also unfavorable for flavone of granularity.The report of more existing microwaves, ultrasound wave auxiliary extraction technology, but apparatus expensive, be difficult to promote.
Summary of the invention
The object of the embodiment of the present invention is to provide a kind of pretreatment of fermenting to improve the method that ginkgetin extracts, and is intended to solve the single alcoholic solution extraction process of the general employing of technology of existing extraction Folium Ginkgo flavone, and the efficiency of extraction is low; The report of more existing microwaves, ultrasound wave auxiliary extraction technology, but apparatus expensive, be difficult to the problem of promoting.
The embodiment of the present invention is to provide a kind of pretreatment of fermenting to improve the method that ginkgetin extracts, and the method comprises the following steps:
Ginkgo leaf powder is broken to 20-60 order;
The Folium Ginkgo mixing that adds water, amount of water 45%-90%;
Mixture, 115 DEG C of sterilizations, packs fermentation tank into;
The weight inoculation strain of pressing Folium Ginkgo in mixture, inoculum concentration is 0.01%-0.02%;
Aerobic fementation 2-11 days under 25-30 DEG C of condition;
Press final argument solid-to-liquid ratio 1:20, ethanol content 60%, adds alcohol and water in right amount, constantly stirs extracting Flavonoid substances;
Filtering suspension liquid, obtains extracting solution;
Extracting solution obtains finished product after vacuum concentration, or is further dried to solid-state.
Further, mixture, 115 DEG C of sterilizations 15 minutes, packs fermentation tank into.
Further, extracting solution through vacuum concentration condition is: 70 DEG C, and 130mbar.
Further, filtering suspension liquid, obtains extracting solution; Extract once again, merge the extracting solution of twice.
Further, fermentation pretreatment can adopt solid-state, also can adopt liquid fermentation.
Further, solid fermentation preprocessing process raw material is only Folium Ginkgo and water; Fermentation condition is: the water content of mixture is 45%-65%, and fermentation temperature is 20-35 DEG C, and fermentation time is 2-11 days.
Further, liquid fermentation preprocessing process raw material is only Folium Ginkgo and water, and fermentation condition is: the water content of mixture is 70-95%, and fermentation temperature is 20-35 DEG C, and fermentation time is 2-11 days.
Further, this fermented bacterium is aspergillosis class fungus, patented strain preserving number CCTCC M2012515.
The fermentation pretreatment that another object of the embodiment of the present invention is to provide above-mentioned improves the application in the method pre-treatment that flavonoid substance extracts within plant tissue that Folium Ginkgo flavone extracts.
The present invention adopts from ancient Semen Ginkgo soil an isolated strain aspergillus niger (patented strain preserving number CCTCC M2012515) to the Folium Ginkgo pretreatment of fermenting, degraded leaf cell, then adopt traditional extracting method to extract, make it to improve the extraction ratio of ginkgetin.With traditional alcoholic solvent extraction method contrast, the flavone extracted amount of Folium Ginkgo improves 20-60%.
Brief description of the drawings
The HPLC finger printing of Fig. 1, flavone provided by the invention: (A) contrast (not fermented sample); (B) the flavone finger printing of patent strain fermentation thing;
Flavones content between Fig. 2, patented strain yeast phase provided by the invention and enzyme activity change;
The systematic evolution tree of Fig. 3, bacterial strain provided by the invention.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
The embodiment of the present invention provides a kind of and has improved for the pretreatment of fermenting the fermented bacterium that Folium Ginkgo flavone extracts, and this fermented bacterium is aspergillosis class fungus, patented strain preserving number CCTCC M2012515.
The patented strain that the present invention uses is to separate and obtain from the soil at the bottom of ancient ginkgo, after mycelia is processed with liquid nitrogen, through milling, DNA extraction, 18SrDNA sequence amplification and order-checking, the sequence of measuring is used NCBI Blast and RDP Chimera Check programs to analyze, and is defined as a strain aspergillus niger (in table 1, this molecular method of this employing of the kind of listed bacterial strain is determined).It is reported first that the present invention applies this fermentation of Aspergillus niger Folium Ginkgo, this aspergillus niger is other bacterial strains relatively, in Folium Ginkgo, growth is vigorous especially, after pretreatment fermentation, the yield of flavone (table 1) of Folium Ginkgo is apparently higher than other bacterial strains, and the component of flavone product after fermentation process consistent with unfermentable product component (ferment in Fig. 2 and the chromatographic peak of fermented sample is not consistent).
The embodiment of the present invention also provides a kind of pretreatment of fermenting to improve the method that ginkgetin extracts, and the method comprises the following steps:
Ginkgo leaf powder is broken to 20-60 order;
The Folium Ginkgo mixing that adds water, amount of water 55%-90%;
Mixture, 115 DEG C of sterilizations 15 minutes, packs fermentation tank into;
The weight inoculation strain of pressing Folium Ginkgo in mixture, inoculum concentration is 0.01%-0.02%;
Aerobic fementation 2-4 days under 25-30 DEG C of condition;
Press final argument solid-to-liquid ratio 1:20, ethanol content 60%, adds alcohol and water in right amount, constantly stirs extracting Flavonoid substances;
Filtering suspension liquid, obtains extracting solution;
Extracting solution obtains finished product after vacuum concentration, or is further dried to solid-state.
In embodiments of the present invention, mixture, 115 DEG C of sterilizations 15 minutes, packs fermentation tank into.
In embodiments of the present invention, extracting solution through vacuum concentration condition is: 70 DEG C, and 130mbar.
In embodiments of the present invention, filtering suspension liquid, obtains extracting solution; Extract once again, merge the extracting solution of twice.
In embodiments of the present invention, fermentation pretreatment can adopt solid-state, also can adopt liquid fermentation.
In embodiments of the present invention, solid fermentation preprocessing process raw material is only Folium Ginkgo and water.Fermentation condition is: the water content of mixture is 45-65%, and fermentation temperature is 26-30 DEG C, and fermentation time is 2-4 days.
In embodiments of the present invention, liquid fermentation preprocessing process raw material is only Folium Ginkgo and water.Fermentation condition is: the water content of mixture is 70-95%, and fermentation temperature is 20-35 DEG C, and fermentation time is 3-6 days.
The method that the embodiment of the present invention provides can be in other plant tissue application in the flavonoid substance pre-treatment of extracting.
In embodiments of the present invention, the fungus and bacterium that strain adopts Penicillium, mucor, reaping hook genus and trichoderma etc. can produce lignocellulose also can be realized effect of the present invention, but effect is poor.
This method is a kind of preprocess method, after processing, can use merely extraction, or in conjunction with ultrasound wave and Microwave-assisted solvent extraction.
Below the general flavone content that extracts after different fungi fermentations and the comparison sheet of cellulose vigor.
The general flavone content extracting after the different fungi fermentations of table 1
* mark the average of identical lower case without significant difference (α=0.05)
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (1)
1. the pretreatment of fermenting improves the method that ginkgetin extracts, and it is characterized in that, the method comprises the following steps:
Ginkgo leaf powder is broken to 20-60 order;
The Folium Ginkgo mixing that adds water, amount of water 45%-90%;
Mixture, 115 DEG C of sterilizations, packs fermentation tank into;
The weight inoculation strain of pressing Folium Ginkgo in mixture, inoculum concentration is 0.01%-0.02%;
Aerobic fementation 2-11 days under 25-30 DEG C of condition;
Press final argument solid-to-liquid ratio 1:20, ethanol content 60%, adds alcohol and water in right amount, constantly stirs extracting Flavonoid substances;
Filtering suspension liquid, obtains extracting solution; Extract once again, merge the extracting solution of twice; Extracting solution obtains finished product after vacuum concentration, or is further dried to solid-state;
Wherein: mixture, 115 DEG C of sterilizations 15 minutes, packs fermentation tank into; Extracting solution through vacuum concentration condition is: 70 DEG C, and 130mbar;
Fermentation pretreatment adopts solid-state or liquid fermentation;
Solid fermentation preprocessing process raw material is only Folium Ginkgo and water; Fermentation condition is: the water content of mixture is 45%-65%, and fermentation temperature is 20-35 DEG C, and fermentation time is 2-11 days;
Liquid fermentation preprocessing process raw material is only Folium Ginkgo and water, and fermentation condition is: the water content of mixture is 70-95%, and fermentation temperature is 20-35 DEG C, and fermentation time is 2-11 days.
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| CN104543004A (en) * | 2014-12-29 | 2015-04-29 | 江苏三益堂保健食品有限公司 | Preparation method and application of bio-fermentation gingko concentrated juice |
| CN106822196B (en) * | 2017-03-23 | 2020-07-07 | 山东天智绿业生物科技有限公司 | Method for simultaneously extracting ginkgo leaf polysaccharide and ginkgo leaf flavone from ginkgo leaves |
| CN107158048A (en) * | 2017-05-12 | 2017-09-15 | 湖南科技学院 | A kind of high efficiency from ginkgo leaf extracts flavones and the method for being converted into glucoside type flavone |
| CN107115367B (en) * | 2017-05-12 | 2020-06-30 | 湖南科技学院 | Fermentation production method for efficiently extracting flavone from ginkgo leaves |
| CN107022590A (en) * | 2017-06-12 | 2017-08-08 | 贵州师范大学 | A kind of method that fermentation of Aspergillus niger honeysuckle improves galuteolin yield |
| CN109363059B (en) * | 2018-10-29 | 2021-11-30 | 南京林业大学 | Method for degrading sensitized and toxic components of ginkgo by utilizing monascus fermentation, product prepared by method and application of product |
| CN109468236A (en) * | 2018-12-27 | 2019-03-15 | 黄冈师范学院 | A method of improving general flavone content in honeysuckle vinasse |
| CN115868590A (en) * | 2021-09-29 | 2023-03-31 | 江苏恒康生物科技有限公司 | Preparation and application of anti-tumor probiotic fermented ginkgo leaf composition |
| CN116570653B (en) * | 2023-04-24 | 2023-10-10 | 遵义市中医院 | Alcohol extraction fermentation process of lipid-lowering traditional Chinese medicine composition and application thereof |
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Application publication date: 20130828 Assignee: Susong Zhong Ming Shan Zhuang Ecological Agriculture Development Co.,Ltd. Assignor: Nanjing Forestry University Contract record no.: 2018320000236 Denomination of invention: Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application Granted publication date: 20141105 License type: Common License Record date: 20181025 |
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