CN103145671B - Application of aspergillus awamori in preparation of quercetin by fermenting lichee peel - Google Patents
Application of aspergillus awamori in preparation of quercetin by fermenting lichee peel Download PDFInfo
- Publication number
- CN103145671B CN103145671B CN201310060543.8A CN201310060543A CN103145671B CN 103145671 B CN103145671 B CN 103145671B CN 201310060543 A CN201310060543 A CN 201310060543A CN 103145671 B CN103145671 B CN 103145671B
- Authority
- CN
- China
- Prior art keywords
- aspergillus awamori
- quercetin
- fructus litchi
- lichee
- peel
- 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.)
- Active
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an application of aspergillus awamori in preparation of quercetin by fermenting lichee peel. Experiments show that quercetin can be extracted from fermented culture after the lichee peel is fermented by aspergillus awamori GIM3.4; but in a control group, no quercetin is detected in the fermented culture obtained after fermentation of lichee peel in which a sterilized aspergillus awamori GIM3.4 spore suspension serving as an inoculator is inoculated, thus the aspergillus awamori GIM3.4 can be used for fermenting the lichee peel to produce quercetin.
Description
Technical field:
The present invention relates to the method obtaining Quercetin from Fructus Litchi, be specifically related to a kind of Aspergillus awamori (Aspergillusawamori) GIM3.4 and prepare application in Quercetin at fermentation Fructus Litchi.
Background technology:
China is the big country producing lichee, and as far back as 1999, only Guangdong Province's Yield of Litchi just reached 680,000 tons, and its dry fruit micromicro reaches more than 20,000 ton (golden shower etc., 2004).In production practice, these more than 20,000 tons of Fructus Litchis are often simply discarded, and have both caused the wasting of resources to be also easily with and have served environmental problem.
Lichee (Litchi chinensis Sonn.) is the Sapindaceae fruit tree in many tropical and subtropical countries (Duan etc. originating in China, 2007b), the pericarp of its shiny red is rich in aldehydes matter (Zhang etc., 2000), the content of total phenol can reach 51.3-102.1g/kg DW(wherein phenolic acid compound 10.3-24.3g/kg DW, total flavones 13.4-37.6g/kg DW, condensed tannin 10.3-32.0g/kgDW) (Wang etc., 2011).In Fructus Litchi, aldehydes matter is of a great variety, different aldehydes matter, and biological activity difference is very large.Therefore, study the composition of Fructus Litchi aldehydes matter and the biological activity of each composition and its aldehydes matter is modified, to obtain the compound with special purpose, having major and immediate significance to the comprehensive utilization of Fructus Litchi.From the document reported, the polyphenol substance in Fructus Litchi can be divided into phenolic acids, anthocyanin class, flavonoid, flavan-3-alcohol class and condensed tannin.Quercetin and derivative thereof are vegitabilia's distribution flavonoid compounds the most widely, have eliminate the phlegm preferably, antitussive action, and have certain antiasthmatic effect, in addition reduce blood pressure in addition, strengthen capillary resistance, reduce capillary fragility, reducing blood-fat, coronary artery dilator, increase the effects such as coronary flow.
Along with the raising of living standards of the people, health care consciousness is also more and more stronger, and the demand of phenols healthcare products in market is very large, but the potential value of Fructus Litchi is not also excavated, so we will make full use of the biologically active substance in Fructus Litchi at present.Can improve from the following aspects: the method 1) finding the aldehydes matter a kind of efficient and convenient, efficient acquisition Fructus Litchi, early stage organic solvent extraction, separation, purifying method can obtain pure aldehydes matter, but consuming time, effort; 2) fermentation substrate can be it can be used as to add in traditional fermented food, as wine, vinegar, soy sauce, to increase the functionally active material of these food; 3) utilize chemistry or biological method to modify aldehydes matter in Fructus Litchi, find novel medicines structure or the prerequisite compound of medicine.Fructus Litchi is the agricultural byproducts being rich in Polyphenols and polysaccharide, can become the source that people obtain biologically active substance.
Quercetin, has another name called quercetin, Quercetin, and its structure, as shown in formula I, is dissolved in Glacial acetic acid, and alkaline aqueous solution is in yellow, and water-soluble hardly, ethanolic soln taste is very bitter.Can be used as medicine, have eliminate the phlegm preferably, antitussive action, and have certain antiasthmatic effect.In addition reduce blood pressure in addition, strengthen capillary resistance, reduce capillary fragility, reducing blood-fat, coronary artery dilator, increase the effects such as coronary flow.Be used for the treatment of chronic bronchitis.Also auxiliary therapeutic action is had to coronary heart disease and hyperpietic.
Summary of the invention:
First object of the present invention is to provide Aspergillus awamori (Aspergillus awamori) GIM3.4 and prepares application in Quercetin at fermentation Fructus Litchi.
The present invention found through experiments, Fructus Litchi is after Aspergillus awamori (Aspergillus awamori) GIM3.4 ferments, Quercetin can be extracted from the fermenting culture after fermentation, and in contrast to be inoculated in Fructus Litchi fermentation using the spore suspension of the Aspergillus awamori of sterilizing (Aspergillusawamori) GIM3.4 as inoculum after fermenting culture in Quercetin do not detected, illustrate that Aspergillus awamori (Aspergillus awamori) the GIM3.4 Fructus Litchi that can ferment produces Quercetin thus.
Second object of the present invention is to provide one and utilizes Aspergillus awamori (Aspergillus awamori) GIM3.4 fermentation Fructus Litchi to produce the method for Quercetin, it is characterized in that, Aspergillus awamori (Aspergillus awamori) GIM3.4 is inoculated in the substratum at least containing Fructus Litchi and water, fermentation culture is carried out, containing Quercetin in fermenting culture after fermentation under the conventional culture conditions of Aspergillus awamori (Aspergillus awamori) GIM3.4.
Can utilize organic solvent extraction, separation, purifying method obtain pure Quercetin.
The described substratum at least containing Fructus Litchi and water preferably presses the mass ratio of 1:1, by the substratum that the Fructus Litchi after pulverizing and water are mixed to get.
Beneficial effect of the present invention is as follows:
1) waste effectively utilizes, and brings larger economic benefit.Fructus Litchi is the Main By product of lichee, and is rich in aldehydes matter, and the present invention Aspergillus awamori GIM3.4 produces new aldehydes matter-Quercetin to after Fructus Litchi fermentation.Along with growth in the living standard, market is very big to the demand dynamics of Quercetin healthcare product, and the present invention effectively utilizes Fructus Litchi to make it produce aldehydes matter Quercetin, thus increases certain income for orchard worker.
2) reaction is quick, and product pollution is little.The present invention is the method for the Quercetin in a kind of efficient and convenient, efficient acquisition Fructus Litchi.Although pure Quercetin can be obtained by the method for organic solvent extraction, separation, purifying, but consuming time, effort, therefore fermentation substrate can be it can be used as when in use to add in traditional fermented food, as wine, vinegar, soy sauce, to increase the functionally active material of these food.
3) approach is new.Not it was reported that at present and brightly can go out Quercetin by extracting directly from Fructus Litchi, the present invention finds, can extract Quercetin in the fermenting culture after Aspergillus awamori (Aspergillus awamori) GIM3.4 ferments to Fructus Litchi.As can be seen here, not extractible aldehydes matter Quercetin in Fructus Litchi, after fermentation, Quercetin is stripping from cell walls, and show stronger DPPH free radical scavenging activity, therefore discharge from plant tissue for research method of microorganism makes to extract aldehydes matter in Fructus Litchi, become extractible aldehydes matter, the prerequisite compound finding novel medicines structure or medicine provides approach.
4) can be practical.Along with the scarcity gradually of world resource, the effective utilization ratio of natural product is also fewer and feweri, utilization of waste material, and the resource that can reach makes full use of; In actual production, efficiently fast from the acquisition Quercetin Fructus Litchi, stronger practicality can be had.The method of research using microbe fermentation is utilized to the polyphenols in agricultural byproducts, the effective ways finding comprehensive development and utilization agricultural byproducts are of great immediate significance.
Aspergillus awamori of the present invention (Aspergillus awamori) GIM3.4 is purchased from Guangdong Province's Culture Collection (GIMCC), address: 5th floor, No. 100, Xianlie Mid Road, Guangzhou City, Guangdong Province, China Guangdong Microbes Inst laboratory building, postcode: 510070, its culture presevation is numbered: GIM3.4.
Embodiment:
Following examples further illustrate of the present invention, instead of limitation of the present invention.
The preparation of the spore suspension of Aspergillus awamori (Aspergillus awamori) GIM3.4:
Using Aspergillus awamori (Aspergillus awamori) GIM3.4 purchased from Guangdong Province's Culture Collection as supplying examination bacterial classification, bacterial classification is containing potato nutrient agar, (formula of often liter of potato nutrient agar is: potato 200g, sucrose 20g, tap water 1000ml and agar 20g, ph nature, compound method: peeling potatoes, be cut into block to add water, the control that 30min(notes firepower is boiled in 121 ° of C sterilizings, can suitably moisturizing), by filtered through gauze, filtrate is with sucrose, supply water to 1000ml, load triangular flask, sterilizing is for subsequent use) culture dish in, cultivate 3 days for 30 DEG C, media surface is rinsed with 20mL0.1% tween 80, make the spore suspension of Aspergillus awamori (Aspergillus awamori) GIM3.4.
Embodiment 1:
1) select fine day to gather, eight to ninety percent ripe, have no mechanical damage and the litchi fruits of disease and pest, after clean water, strip pericarp by hand, naturally dry, pulverize with plant pulverizer, the sieve of mistake 1mm is placed in dry environment to be preserved, and obtains Fructus Litchi powder;
2) get 30 2000mL triangular flasks and put into 100g Fructus Litchi powder respectively, then add the distilled water of 100mL, as fermention medium, 121 DEG C of sterilizing 20min.The spore suspension getting the above-mentioned Aspergillus awamori of 1mL (Aspergillus awamori) GIM3.4 adds in the fermention medium of every bottle, after mixing, cultivates 6 days, within every 2 days, stirs substratum, obtain fermenting culture for 30 DEG C.
3) 500mL volume fraction 60% aqueous ethanolic solution is added in every bottle of fermenting culture, 210W supersound extraction 30min, after filtration, then extract once by identical condition, merging filtrate, 50 DEG C of concentrating under reduced pressure, remove ethanol, with 200mL extraction into ethyl acetate 2 times, merge twice extraction liquid, 50 DEG C of evaporated under reduced pressure, obtain ethyl acetate extract.
4) get above-mentioned ethyl acetate extract 45g add a little methyl alcohol make it dissolve, add 65g silica gel mixed sample, leave standstill 30min, evaporated under reduced pressure, and use mortar grinder sample, sample is added on silicagel column (5inner diameter × 130cm), with the chloroform/methanol (CHCl of different volumes ratio
3/ CH
3oH10:0,9.8:0.2,9.6:0.4,9.2:0.8,9:1) gradient with the flow velocity wash-out of 15mL/min, each gradient elution 7.5L moving phase.CHCl is collected with 500ml Erlenmeyer flask
3/ CH
3oH is the eluted product under 9.6:0.4 gradient, collects 300mL for every bottle.Merge similar compositions (developping agent is volume ratio toluene: chloroform: acetone=40:25:35, the Rf=0.55 of Quercetin) by tlc, then use Sephadex LH-20(1 × 170cm), using methyl alcohol as moving phase wash-out, be further purified.The preliminary purification sample decompression obtained is steamed to 2mL, be prepared with partly preparing reverse high performance liquid chromatography (PRC-ODS column25 × 460mm) again, reverse preparation take methanol-water as the flow rate gradient of moving phase, 5ml/min, and observes out peak situation under 280nm.Methanol-water initial proportion is volume ratio 40:60(methyl alcohol: water) wash-out 10min, then use volume ratio 45:55(methyl alcohol: water) wash-out 20min, then methanol ratio is risen to the reverse post of 100% flushing.According to 45:55(methyl alcohol: water) go out peak situation under gradient, collect eluted product (appearance time 18-20min, note: be preparation liquid phase, therefore peak width is wider) under this gradient, obtain sterling compound 1.
Compound 1 is yellow powder.Under positive and negative ion pattern, ESI-MS analyzes known, and compound 1 has m/z303.0 [M+H] respectively
+, m/z301.0 [M-H]
-quasi-molecular ion peak, can judge that the molecular weight of this chemical combination is as 302.
1h NMR and
13the data of C NMR are in table 1.Have 1 hydrogen atom to be that 7.60ppm presents a double doublet in chemical shift as seen from table, lotus root close constant be on 2.07Hz(phenyl ring between position carbon connects hydrogen
4j lotus root close) and 8.5Hz(phenyl ring on adjacent carbon
3j lotus root is closed), containing the structure unit that aromatic ring 1,3,4 replaces in above data declaration F4.Separately have 2 hydrogen atoms to be that 6.15ppm and 6.35ppm place presents two groups of doublets at chemical displacement value respectively, and lotus root close constant be respectively 1.8 with on 1.9Hz(phenyl ring between position carbon is connected hydrogen
4j lotus root is closed).These meet the feature of the upper hydrogen of C-6, C-8 on C-5 and C-7 disubstituted flavonoid compound A ring.Composed and the data of DEPT collection of illustrative plates from carbon, compound F 17-hydroxy-corticosterone 4 always has 15 carbon, wherein has 9 quaternary carbons, and does not have methylene radical (CH in this compound molecule
2).Chemical displacement value be 117.2 and its peak height of 132.2ppm place be about 2 times of other peak height, can be judged as there are 2 carbon respectively.And containing a carbonyl carbon (δ 177.3) in molecule.
Table 1, compound 1
1h,
13c data (deuterated methanol is solvent)
According to the data of HPLC, ESI-MS and NMR, compound 1 is accredited as Quercetin.As can be seen here, Quercetin output can be obtained with aforesaid method from Fructus Litchi powder and reach 50 μ g/mg Fructus Litchi powder.
Comparative example 1:
1) select fine day to gather, eight to ninety percent ripe, have no mechanical damage and the litchi fruits of disease and pest, after clean water, strip pericarp by hand, naturally dry, pulverize with plant pulverizer, the sieve of mistake 1mm is placed in dry environment to be preserved, and obtains Fructus Litchi powder.
2) get 100g Fructus Litchi powder and be placed in 2000mL triangular flask, add the distilled water of 100mL, as fermention medium, 121 DEG C of sterilizing 20min.The spore suspension getting Aspergillus awamori (Aspergillus awamori) GIM3.4 of 1mL sterilizing adds in fermention medium, after mixing, 30 DEG C cultivate 6 days, within every 2 days, stir substratum, obtain fermenting culture, do 2 parallel.
The extraction of Quercetin in fermenting culture in comparative example, analytical procedure, with embodiment 1, does not detect containing Quercetin in the fermenting culture in result display comparison example.
Claims (2)
1. one kind utilizes Aspergillus awamori (Aspergillus awamori) GIM 3.4 Fructus Litchi that ferments to produce the method for Quercetin, it is characterized in that, Aspergillus awamori (Aspergillus awamori) GIM 3.4 is inoculated in the substratum at least containing Fructus Litchi and water, fermentation culture is carried out, containing Quercetin in fermenting culture after fermentation under the conventional culture conditions of Aspergillus awamori (Aspergillus awamori) GIM 3.4.
2. method according to claim 1, is characterized in that, the described substratum at least containing Fructus Litchi and water is the mass ratio by 1:1, by the substratum that the Fructus Litchi after pulverizing and water are mixed to get.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310060543.8A CN103145671B (en) | 2013-02-26 | 2013-02-26 | Application of aspergillus awamori in preparation of quercetin by fermenting lichee peel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310060543.8A CN103145671B (en) | 2013-02-26 | 2013-02-26 | Application of aspergillus awamori in preparation of quercetin by fermenting lichee peel |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103145671A CN103145671A (en) | 2013-06-12 |
| CN103145671B true CN103145671B (en) | 2015-03-04 |
Family
ID=48544049
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310060543.8A Active CN103145671B (en) | 2013-02-26 | 2013-02-26 | Application of aspergillus awamori in preparation of quercetin by fermenting lichee peel |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103145671B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107468533A (en) * | 2017-08-22 | 2017-12-15 | 广州聚注通用技术研究院有限公司 | Saussurea involucrata Quercetin with radiation-resisting whitening activity and its preparation method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1483825A (en) * | 2003-08-01 | 2004-03-24 | 金凤燮 | Method for preparing isoquercetin and quercetin by enzymatic method and hydrolyzing rutin |
| EP2438916A1 (en) * | 2009-06-03 | 2012-04-11 | Fuji-Sangyo Co., Ltd. | Neurite elongation agent, memory-improving agent and anti-alzheimer agent comprising 4'-demethylnobiletin or 4'-demethyltangeretin as active ingredient, and process for production of the compound |
| CN102895388A (en) * | 2012-09-28 | 2013-01-30 | 桂林益元素生物科技有限公司 | Lychee polyphenol extracts and extraction method thereof |
-
2013
- 2013-02-26 CN CN201310060543.8A patent/CN103145671B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1483825A (en) * | 2003-08-01 | 2004-03-24 | 金凤燮 | Method for preparing isoquercetin and quercetin by enzymatic method and hydrolyzing rutin |
| EP2438916A1 (en) * | 2009-06-03 | 2012-04-11 | Fuji-Sangyo Co., Ltd. | Neurite elongation agent, memory-improving agent and anti-alzheimer agent comprising 4'-demethylnobiletin or 4'-demethyltangeretin as active ingredient, and process for production of the compound |
| CN102895388A (en) * | 2012-09-28 | 2013-01-30 | 桂林益元素生物科技有限公司 | Lychee polyphenol extracts and extraction method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Enhanced DPPH radical scavenging activity and;Sen Lin等;《Chemistry Central Journal》;20121231 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103145671A (en) | 2013-06-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103263448B (en) | Fermentation bacteria used for fermentation pretreatment to improve extraction of Ginkgo biloba L. leaf flavone and application | |
| CN106265846A (en) | A kind of fermentation process improving Radix Rhodiolae oxidation resistance | |
| CN104306443A (en) | Method for comprehensively utilizing cinnamomum longepaniculatum leaves | |
| CN103655215A (en) | Paecilomyces varioti extract with tyrosinase activity and scavenging free radical activity and application thereof | |
| CN104593441A (en) | Method for simultaneously extracting volatile oil, polysaccharides and amino acids from tricholoma matsutake | |
| Gao-Sheng et al. | Accumulation of biomass and four triterpenoids in two-stage cultured Poria cocos mycelia and diuretic activity in rats | |
| CN106086147A (en) | Fungal elicitor is utilized to induce the method extracting betulic acid from Inonqqus obliquus | |
| CN111096983A (en) | Phellinus igniarius phenolic extract with hypoglycemic activity and preparation and application thereof | |
| CN104983915A (en) | Preparation method of natural lycium ruthenicum composite antioxidant | |
| CN108102928B (en) | One plant of gingko endogenous fungus and its application | |
| CN103880826B (en) | A kind of isobenzofuran ketonic compound and its preparation method and application | |
| CN103145671B (en) | Application of aspergillus awamori in preparation of quercetin by fermenting lichee peel | |
| CN103467262B (en) | Method for preparing 9-oxonerolidol from camphor tree plants | |
| CN115340952A (en) | Method for improving quality of fermentation mycelium of phyllobacterium ribrum by using honeysuckle stem extract | |
| CN108640956A (en) | A method of preparing flavonoid glycoside from tea seed | |
| KR20120053564A (en) | Fermented ginseng with ganoderma lucidum beverage manufacturing method and that beverage | |
| JP4852353B2 (en) | Novel cinnamic acid derivative, process for producing the same and propolis fermented product | |
| CN102633806A (en) | Extraction and separation method of harringtonine | |
| Ho et al. | The hepatoprotective activity against ethanol-induced cytotoxicity by aqueous extract of Antrodia cinnamomea | |
| CN104593443A (en) | Preparation method of agilawood chromone component | |
| CN104894184B (en) | A kind of Agricus blazei liquid fermentation medium, zymotechnique and fermented product adding yam skin | |
| CN103820330B (en) | One strain variable color aspergillus and preparing the application in podophyllotoxin list glucoside and podophyllotoxin | |
| CN109288902B (en) | Preparation method and application of wild jujube leaf total flavone fermentation product with strong antioxidant activity | |
| CN108440477B (en) | A method of preparing Quercetin from tungoiltree juvenile leaf | |
| KR100857687B1 (en) | A method for isolation of high concentration of magnesium lithospermate b from salviae miltiorrhizae |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |