CN102511367B - Method of inducing soybeans to generate glyceollin - Google Patents
Method of inducing soybeans to generate glyceollin Download PDFInfo
- Publication number
- CN102511367B CN102511367B CN2011104500648A CN201110450064A CN102511367B CN 102511367 B CN102511367 B CN 102511367B CN 2011104500648 A CN2011104500648 A CN 2011104500648A CN 201110450064 A CN201110450064 A CN 201110450064A CN 102511367 B CN102511367 B CN 102511367B
- Authority
- CN
- China
- Prior art keywords
- soybean
- glyceollin
- alginic acid
- kernel
- sterile water
- 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
Images
Classifications
-
- Y02P60/216—
Abstract
The invention discloses a method of inducing soybeans to generate glyceollin, which includes steps: firstly, pretreating soybean seeds to expand and husking the seeds so as to obtain soybean cotyledons; and secondly, spraying alginic acid oligosaccharide solution to the surfaces of the soybean cotyledons, and culturing to obtain soybeans containing glyceollin. The alginic acid oligosaccharide is used for exogenous induction, so that content of glyceollin in the soybeans is increased, a novel method for deep exploration, development and utilization of soybean seed sources and high-added-value secondary metabolites of the soybean seeds is provided, and a novel way for improving comprehensive utilization value of the alginic acid oligosaccharide is exploited. The glyceollin content of the glyceollin-enriched soybean powder of fresh soybeans prepared by the method is up to 0.5mg per gram of fresh soybeans, and namely the soybean glyceollin content is 0.54mg per gram of dried soybeans.
Description
Technical field
The present invention relates to the method that a kind of inducing soybean produces glyceollin.
Background technology
Soybean (Glycine max) is a pulse family Glycine annual herb plant, and nutritive value is the highest in beans.Except soybean itself intrinsic isoflavonoid have the various biological effect, soybean under stress can produce a kind of induced product glyceollin (glyceollins), structural formula is suc as formula shown in the I-III; It can efficiently block the growth and the diffusion of gynecological tumor.Existing result of study shows that glyceollin can suppress 53.4%MCF-7 tumor propagation and 73.1%BG-1 tumor propagation.In addition, glyceollin can suppress the expression of progesterone receptor in the MCF-7 cell that oestrogenic hormone induces fully, and part suppresses its expression at the BG-1 cell, blocks growth of tumor and diffusion with this.Glyceollins might become the another healthy element in the soybean.
(alginate oligosaccharides, AOS) mainly by adopting chemistry or biological method degraded algin (algin) to obtain, algin extensively is present in the brown alga alginic acid oligosaccharides, is one of filler of brown alga cell wall.The alginic acid oligosaccharides is for promoting body growth, improve the immunity of organism defence capability that the invasion that reduces disease has special effect.
Summary of the invention
The purpose of this invention is to provide a kind of method that obtains glyceollin by the exogenous stimulation soybean.
Method provided by the present invention is to be that derivant, soybean are raw material with the alginic acid oligosaccharides, by soaking, induce, cultivate, make the soybean of being rich in glyceollin.
Concrete preparation method comprises the steps:
1) soybean kernel is carried out preliminary treatment, make its expansion and remove kind of a skin, obtain soybean cotyledon;
2), obtain containing the soybean of glyceollin at described soybean cotyledon surface sprinkling alginic acid oligosaccharide solution and cultivate.
Wherein, soybean kernel described in the step 1) specifically can be selected from following any one: soya bean seed, green soya bean seed and black soya bean seed.Wherein, the soya bean seed comprises transgenosis soya bean seed and non-transgenic soya bean seed.
It is specific as follows that soybean kernel is carried out pretreated method: be that 75% ethanolic solution carries out sterilization for soybean kernel with volume content at first, then to the soybean kernel sterile water drip washing after the sterilization, then soybean kernel is soaked in sterile water, the soybean kernel that will soak at last after expanding is sloughed kind of a skin.
In the process of above-mentioned sterilization, soybean kernel and volume content are that the proportioning of 75% ethanolic solution can be 1g: 3-5ml, and the time of sterilization can be 3-5min;
In the process of above-mentioned drip washing, the proportioning of soybean kernel and sterile water can be 1g: 3-5ml, the number of times of drip washing can be 2-3 time;
In the process of above-mentioned immersion, the proportioning of soybean kernel and sterile water is 1g: 3-5ml, and the time of immersion is 3-8h.
Step 2) oligosaccharides of alginic acid described in is to be the alginic acid oligosaccharides of 2-10 by algin through the degree of polymerization that algin catenase degraded or acid degradation obtain.
Step 2) oligosaccharide solution of alginic acid described in is specially the alginic acid oligosaccharide solution that mass concentration is 1%-15%;
When soybean cotyledon surface sprinkling alginic acid oligosaccharide solution, the proportioning of soybean cotyledon and alginic acid oligosaccharide solution is 1g: 3mL-20mL.Better to induce effect in order reaching, before spraying the alginic acid oligosaccharide solution, need to mark some roads wound on the soybean cotyledon surface, as mark 2-3 road wound.
To surface sprinkling the soybean cotyledon of alginic acid oligosaccharide solution to carry out culture condition as follows: under 20 ℃-40 ℃ of temperature, relative moisture 30%-70% condition, cultivated 1-7 days in the dark.
In the process of cultivating, soybean cotyledon places the culture dish that the sterilization filter paper that soaks with sterile water is housed.
In order to obtain being rich in the soybean meal of glyceollin, described method comprises: with step 2) soybean that contains glyceollin that obtains is after being dried to moisture under 90 ℃-120 ℃ the condition and being lower than 5%, and the step of pulverizing.
It is heavy that glyceollin content in the resulting soybean meal that contains glyceollin can reach the bright beans of 0.2-0.5mg/g.
The present invention induces by adopting alginic acid oligosaccharides external source, improve the content of glyceollin in the soybean, for deeply excavation and development and use soya seeds resource and its high added value secondary metabolite provide a kind of new method, also for improving the comprehensive utilization value of alginic acid oligosaccharides, open up a new approach simultaneously.Glyceollin content in the soybean meal that is rich in glyceollin of employing the inventive method preparation can reach the bright beans of 0.5mg/g and weighs.It can be used as to produce is rich in the raw material of glyceollin functional food, or can be used as feed addictive and be used for cultivated animals such as livestock and poultry, aquatic product and ruminate feed such as cultivated animals, also can be used for preparing the glyceollin one-component.
Description of drawings
Fig. 1 measures spectrogram for the HPLC without glyceollin glyceollins in the common soybeans of inducing.
Fig. 2 measures spectrogram for the HPLC of glyceollin glyceollins in the soybean that the alginic acid oligosaccharides is induced.
Embodiment
Below by specific embodiment method of the present invention is described, but the present invention is not limited thereto.
Experimental technique described in the following embodiment if no special instructions, is conventional method; Described reagent and material if no special instructions, all can obtain from commercial channels.
The alginic acid oligosaccharides is available from Dalian Glycobio Co., Ltd, and food-grade alginic acid oligosaccharides, content be more than 90%, degree of polymerization 2-9.
The soybean meal of glyceollin is rich in embodiment 1, preparation
Selected soya bean seed (water content≤1%, actual is 0.77%) 10g, be that 75% ethanolic solution is according to 1g: 3ml (soybean weight: dosage sterilization 5min volumes of aqueous ethanol) with volume content, (soybean weight: the sterile water volume) the sterile water drip washing of dosage is 2 times, then at 1g: 3ml (soybean weight: the sterile water volume) soak 5h in the sterile water of volume to use 1g: 3ml again.The soybean of soaking after expanding is sloughed kind of a skin, be divided into two, and on the beans sheet, mark 2-3 road wound, the sprinkling mass concentration is 1% alginic acid oligosaccharide solution 15mL, the soybean cotyledon that sprayed the alginic acid oligosaccharide solution is put into the culture dish that the sterilization filter paper that soaks with sterile water is housed, under 20 ℃ of temperature, relative moisture 50% condition, cultivated 7 days in the dark.With the soybean after cultivating 90 ℃-120 ℃ be dried to moisture and be lower than 5% after, be ground into powder.The ratio that adds the ethanolic solution of 3.0mL volume content 80% according to the 1.0g soybean meal is extracted glyceollin (glyceollins), and 50 ℃ of heated and stirred 1h are cooled to room temperature, centrifugal 10min under the 12000rpm condition, draw supernatant,, carry out HPLC and measure through 0.22 μ m micro-pore-film filtration.
The HPLC condition determination is: the Waters2695 chromatographic system, join Waters PD UV-detector, and pillar is C
18Reverse-phase chromatographic column (4.6 * 250mm, 5 μ m), column temperature: 40 ℃, adopt binary flowing phase: the A=mass concentration is 0.1% acetic acid aqueous solution (pH=3.0); The B=100% acetonitrile.Flow velocity: 1.0mL/min, gradient elution: 0-17min, 0% acetonitrile → 45% acetonitrile; 17min-27min, 45% acetonitrile → 90% acetonitrile; 27min-33min, 90% acetonitrile, 33min holds time; Detect wavelength: 285nm, sample size 10 μ L.The peak retention time that goes out of glyceollin glyceollins is 23-24min, and chromatogram as shown in Figure 2.Measure through the HPLC method, the glyceollin content in the soybean meal that is rich in glyceollin of present embodiment preparation is that the bright beans of 0.25mg/g are heavy, and promptly glyceollin content is that the dried beans of 0.27mg/g are heavy.Can contain glyceollin hardly in the undressed common soya bean seed.(Figure 1 shows that the HPLC testing result of glyceollin content in the undressed common soya bean, from spectrogram, as can be seen, do not detect glyceollin in the common soya bean.)
The soybean meal of glyceollin is rich in embodiment 2, preparation
Selected soya bean seed (water content≤1%, actual is 0.77%) 10g, be that 75% ethanolic solution is according to 1g: 5ml (soybean weight: dosage sterilization 3min volumes of aqueous ethanol) with volume content, (soybean weight: the sterile water volume) the sterile water drip washing of dosage is 3 times, then at 1g: 5ml (soybean weight: the sterile water volume) soak 6h in the sterile water of volume to use 1g: 5ml again.The soybean of soaking after expanding is sloughed kind of a skin, be divided into two, and on the beans sheet, mark 2-3 road wound, the sprinkling mass concentration is 4% alginic acid oligosaccharide solution 5mL, the soybean cotyledon that sprayed the alginic acid oligosaccharide solution is put into the culture dish that the sterilization filter paper that soaks with sterile water is housed, under 25 ℃ of temperature, relative moisture 60% condition, cultivated 5 days in the dark.With the soybean after cultivating 90 ℃-120 ℃ be dried to moisture and be lower than 5% after, be ground into powder.Extract glyceollin and measure its content according to the method among the embodiment 1 with the HPLC method.The result shows that the glyceollin content in the soybean meal that is rich in glyceollin of present embodiment preparation is that the bright beans of 0.5mg/g are heavy, and promptly glyceollin content is that the dried beans of 0.54mg/g are heavy.
The soybean meal of glyceollin is rich in embodiment 3, preparation
Selected soya bean seed (water content≤1%, actual is 0.77%) 10g, be that 75% ethanolic solution is according to 1g: 4ml (soybean weight: dosage sterilization 4min volumes of aqueous ethanol) with volume content, (soybean weight: the sterile water volume) the sterile water drip washing of dosage is 2 times, then at 1g: 4ml (soybean weight: the sterile water volume) soak 8h in the sterile water of volume to use 1g: 4ml again.The soybean of soaking after expanding is sloughed kind of a skin, be divided into two, and on the beans sheet, mark 2-3 road wound, spray the alginic acid oligosaccharide solution 3mL of 8% concentration, the soybean cotyledon that sprayed the alginic acid oligosaccharide solution is put into the culture dish that the sterilization filter paper that soaks with sterile water is housed, under 35 ℃ of temperature, relative moisture 70% condition, cultivated 4 days in the dark.With the soybean after cultivating 90 ℃-120 ℃ be dried to moisture and be lower than 5% after, be ground into powder.Extract glyceollin and measure its content according to the method among the embodiment 1 with the HPLC method.The result shows that the glyceollin content in the soybean meal that is rich in glyceollin of present embodiment preparation is that the bright beans of 0.38mg/g are heavy, and promptly glyceollin content is that the dried beans of 0.41mg/g are heavy.
Claims (8)
1. the method for an inducing soybean generation glyceollin comprises the steps:
1) soybean kernel is carried out preliminary treatment, make its expansion and remove kind of a skin, obtain soybean cotyledon;
2), obtain containing the soybean of glyceollin at described soybean cotyledon surface sprinkling alginic acid oligosaccharide solution and cultivate;
Step 2) oligosaccharides of alginic acid described in is to be degraded or acid degradation through algin catenase by algin, and the degree of polymerization that obtains is the alginic acid oligosaccharides of 2-10;
Step 2) mass concentration of alginic acid oligosaccharides is 1%-15% in the oligosaccharide solution of alginic acid described in; The proportioning of described soybean cotyledon and alginic acid oligosaccharide solution is 1g:3mL-20mL;
Draw on described soybean cotyledon surface some roads wound;
Described culture condition is as follows: cultivated 1-7 days under 20 ℃-40 ℃ of temperature, relative moisture 30%-70% condition in the dark.
2. method according to claim 1 is characterized in that: draw on described soybean cotyledon surface 2-3 road wound.
3. method according to claim 1 and 2 is characterized in that: in the process of described cultivation, the soybean cotyledon that the alginic acid oligosaccharide solution is crossed in surface sprinkling places the culture dish that the sterilization filter paper that soaks with sterile water is housed.
4. method according to claim 1 and 2 is characterized in that: described soybean kernel is selected from any one in the following seed: soya bean seed, green soya bean seed and black soya bean seed.
5. method according to claim 1 and 2, it is characterized in that: it is as follows in the step 1) soybean kernel to be carried out pretreated method: be that 75% ethanolic solution carries out sterilization for soybean kernel with volume content at first, then to the soybean kernel sterile water drip washing after the sterilization, then soybean kernel is soaked in sterile water, the soybean kernel that will soak at last after expanding is sloughed kind of a skin.
6. method according to claim 5 is characterized in that: in the process of described sterilization, soybean kernel and volume content are that the proportioning of 75% ethanolic solution is 1g:3-5ml, and the time of sterilization is 3-5min;
In the process of described drip washing, the proportioning of soybean kernel and sterile water is 1g:3-5ml, and the number of times of drip washing is 2-3 time;
In the process of described immersion, the proportioning of soybean kernel and sterile water is 1g:3-5ml, and the time of immersion is 3-8h.
7. method according to claim 1 and 2, it is characterized in that: described method also comprises: with step 2) soybean that contains glyceollin that obtains is after being dried to moisture under 90 ℃-120 ℃ the condition and being lower than 5%, pulverize, obtain containing the soybean meal of glyceollin.
8. method according to claim 7 is characterized in that: the glyceollin content in the described soybean meal that contains glyceollin is that the bright beans of 0.2-0.5mg/g are heavy.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104500648A CN102511367B (en) | 2011-12-29 | 2011-12-29 | Method of inducing soybeans to generate glyceollin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104500648A CN102511367B (en) | 2011-12-29 | 2011-12-29 | Method of inducing soybeans to generate glyceollin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102511367A CN102511367A (en) | 2012-06-27 |
| CN102511367B true CN102511367B (en) | 2013-07-24 |
Family
ID=46282675
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011104500648A Active CN102511367B (en) | 2011-12-29 | 2011-12-29 | Method of inducing soybeans to generate glyceollin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102511367B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170065554A1 (en) * | 2014-02-20 | 2017-03-09 | MicroBlome Therapeutics LLC | Activated soy pod fiber |
| CN105453918A (en) * | 2015-12-03 | 2016-04-06 | 丁玉琴 | Method for preparing glyceollin by means of induction effect of konjak composite oligosaccharide |
| CN108660099B (en) * | 2018-07-05 | 2021-06-25 | 中国农业科学院饲料研究所 | Bacillus amyloliquefaciens for inducing soybeans to generate glyceollin and application thereof |
-
2011
- 2011-12-29 CN CN2011104500648A patent/CN102511367B/en active Active
Non-Patent Citations (6)
| Title |
|---|
| Alginate-deriving oligosaccharide production by alginase from newly isolated Flavobacterium sp. LXA and its potential application in protection against pathogens;An Q D等;《Journal of Applied Microbiology》;20091231;第106卷(第1期);161-170 * |
| An Q D等.Alginate-deriving oligosaccharide production by alginase from newly isolated Flavobacterium sp. LXA and its potential application in protection against pathogens.《Journal of Applied Microbiology》.2009,第106卷(第1期),161-170. |
| 乞永艳等.大豆中Glyceollins诱导和累积研究进展.《大豆科学》.2007,第26卷(第3期),400-406. |
| 大豆中Glyceollins诱导和累积研究进展;乞永艳等;《大豆科学》;20070630;第26卷(第3期);400-406 * |
| 王媛媛等.褐藻寡糖的生物活性与应用研究进展.《食品与发酵工业》.2010,(第10期),122-126. |
| 褐藻寡糖的生物活性与应用研究进展;王媛媛等;《食品与发酵工业》;20100910(第10期);122-126 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102511367A (en) | 2012-06-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102007876B (en) | Biological enzyme fish guano and preparation method thereof | |
| CN103120135A (en) | Pollution-free pelteobagrus fulvidraco breeding method | |
| CN102511367B (en) | Method of inducing soybeans to generate glyceollin | |
| CN104351091A (en) | Black carp breeding technology | |
| CN104489293A (en) | Lactating cattle feed and preparation method thereof | |
| CN107821243A (en) | The cultural method of fresh water perch | |
| CN105613006A (en) | Peanut planting method | |
| CN103222439A (en) | Grading farming method of specific pathogen free seed shrimp of penaeus vanmamei | |
| CN107473277A (en) | A kind of water quality of aquaculture pond cleanser and preparation method thereof | |
| CN104770320A (en) | Freshwater shrimp high-yield aquaculture method | |
| CN104304829A (en) | Grass carp feed | |
| CN104186431A (en) | High-density artemia breeding method with single-cell protein single-step food chain utilized | |
| CN105706974A (en) | Rice field fish culture method | |
| CN105900885A (en) | Culture method of ecological crucian | |
| CN103718994B (en) | A kind of method that artificial induction pseudorasbora parva parent population sexual gland is synchronously grown | |
| CN104664065A (en) | Supplementary feeding formula of under-forest randomly rearing chickens suitable for nursery stock forest land, and raising method of under-forest randomly rearing chickens | |
| CN106007880A (en) | Preparation method of marigold seedling raising substrate | |
| CN109874446A (en) | A kind of breeding method of Queensland nut seed | |
| CN105076819A (en) | Raising feed special for crucian and preparation method thereof | |
| CN106561502A (en) | Fry rearing method | |
| CN107439478A (en) | A kind of black pig ecological cultivation method | |
| CN104285863B (en) | A kind of method inducing gynogenesis Nibea albiflora puppet milter | |
| CN110199751A (en) | A kind of method for culturing seedlings of tea tree | |
| CN108935257A (en) | A kind of shrimp ponds net cage combined breeding method | |
| CN107667937A (en) | A kind of feeding method of cray |
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 |