CN102648757A - Preparation method of fermented soybeans with alpha-glucosaccharase inhibitory activity - Google Patents
Preparation method of fermented soybeans with alpha-glucosaccharase inhibitory activity Download PDFInfo
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- CN102648757A CN102648757A CN2012101469224A CN201210146922A CN102648757A CN 102648757 A CN102648757 A CN 102648757A CN 2012101469224 A CN2012101469224 A CN 2012101469224A CN 201210146922 A CN201210146922 A CN 201210146922A CN 102648757 A CN102648757 A CN 102648757A
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Abstract
The invention relates to a preparation method of fermented soybeans with alpha- glucosaccharase inhibitory activity, which is characterized in that: soybeans bought from market are adopted as raw materials, a py2(bacillus subtilis py2) which is a strain separated from traditional fermented food is used for having the pure solid fermentation so as to acquire the fermented food with the alpha-glucosaccharase inhibitory activity. The preparation method is simple to operate, and the prepared fermented food has stable alpha-glucosaccharase inhibitory activity and is hopefully used as food with a hyperglycemic function.
Description
Technical field
The present invention relates to a kind of preparation method that alpha-glucosidase suppresses active fermentation food that has, belong to food processing field, utilize alpha-glucosidase restrainer to produce bacterium and ferment with grain raw materials such as soybean, rice, corns.
Technical background
Alpha-glucosidase restrainer is because of having and monose or the similar chemical constitution of oligosaccharides; Can the competitive activity that suppresses the carbohydrate inversion enzyme; Thereby can effectively control diabetic's level of postprandial blood sugar; Postpone the use of insulin, in diabetic's treatment and healthcare process, occupy critical role.In addition, alpha-glucosidase restrainer can also CKIs and lipid glycosylation, has effects such as anticancer, anti HIV-1 virus, anti-murine leukemia virus, antiatherosclerosis.
In recent years,, adopt dietetic treatment, regularly take food with function of reducing blood sugar has obtained countries in the world as the supplemental treatment regimens of treating diabetes approval along with riseing year by year of diabetic's quantity.According to IDF (
Http:// www.idf.org) the recent statistics data showed 2010 years: worldwide nearly 2.85 hundred million diabetics in 2009, diabetes have become the whole world the 4th and have caused dead disease greatly.At present existing more than 9,000 ten thousand diabetics (latest data in 2010) of China.Wholistic therapy schemes such as routing motion treatment, drug therapy help reducing the onset diabetes rate and the death rate on the dietetic treatment basis.Therefore development of new, inexpensive, effectively can to prevent or treat the functional food of diabetes significant.
Although more and more deep to alpha-glucosidase restrainer research, most of research belongs to field of medicaments, and is less to the food research that contains alpha-glucosidase restrainer.Contain alpha-glucosidase restrainer in part animal and plant resource such as mulberry leaf, eggplant, the silkworm etc.; Can be used as the raw material of hypoglycemic food; But these resources receive the restriction of conditions such as cultivated area, output, cultivation cycle usually, exist raw material to obtain problems such as cycle length, production cost height.And directly can partly solve with the animal and plant resource as the source of function of blood sugar reduction food with the fermented food that contains alpha-glucosidase restrainer is that raw material production the cycle grows up to this high problem when containing alpha-glucosidase restrainer food.On the one hand, can well control the quality of final products through the control fermentation condition; On the other hand, can directly shorten the production cycle, reduce production costs through microbial fermentation.
Have the history of the fermented soya bean of integration of drinking and medicinal herbs function, discover that the Chinese traditional fermented food fermented soya bean of part brand have alpha-glucosidase and suppress active as the existing more than one thousand years of traditional food of China's adjuvant therapy of diabetes.But because its zymotechnique of fermented soya bean, the fermented bacterium of different manufacturers production have nothing in common with each other, the fermented soya bean that only lack the part brand have alpha-glucosidase and suppress active; Even same producer, it is also unstable that its alpha-glucosidase of the fermented soya bean that different batches is produced suppresses active height, even when occurring and have alpha-glucosidase to suppress active, the time and do not have alpha-glucosidase to suppress active problem.Suppressing active problem of unstable for how solving the fermented soya bean alpha-glucosidase, obtaining to have the research that stable alpha-glucosidase suppresses active fermented soya bean, also rarely have report at present.
It is a kind of that the object of the invention is developed to the problems referred to above just is that feedstock production has the method that stable alpha-glucosidase suppresses active fermented soybean food with commercially available soybean.This method is so that strain strain separated from the traditional zymotic soy food product---bacillus subtilis Py2 is initial bacterial strain; With the soaking soybean is raw material; Through controlling its fermentation temperature, humidity and fermentation time; The fermented soybean food of preparation stay in grade, this product have stable and strong alpha-glucosidase and suppress active.
The present invention adopts pure-blood ferment, and fermentation period is short, and fermentation condition is easy to control; Reduced and contained the cost that alpha-glucosidase suppresses active raw-food material; Shortened the production cycle, and product with stable quality, have a good application prospect in function of blood sugar reduction development of food field.
Summary of the invention
The technical problem that the present invention solved is, is that raw material has the method that stable alpha-glucosidase suppresses the fermented soybean food of activity through the microbial fermentation processes preparation with commercially available soybean.
The objective of the invention is to realize through following scheme:
1. one kind has the fermented soybean food that alpha-glucosidase suppresses activity, and it is characterized in that: with soybean or other edible grain is raw material, is fermentation strain with bacillus subtilis Py2, forms through pure-blood ferment.
2. fermented soybean food as claimed in claim 1, it is characterized in that: the preparation method may further comprise the steps: soaking soybean, sterilization, inoculation, fermentation, fermentation condition is: temperature 20-40 ℃, humidity 70-95%, fermentation time are 12-96h.
3. fermented soybean food as claimed in claim 2; It is characterized in that: said immersion raw material and sterilization steps are: the water that soybean adds 3 times of quality soaks 12-24h down at 10-40 ℃; Place high-pressure sterilizing pot, obtain culture matrix in 121 ℃ of boiling 15-60min.
4. fermented soybean food as claimed in claim 2 is characterized in that: said inoculation step is: bacterial strain is carried out activation, get glycerine and guarantee in the inoculum concentration access activation medium of bacterial classification by 1% (v:/v) of Tibetan; Medium component is 1% beef extract-peptone; 0.5% yeast extract, 1% sodium chloride is cultivated 16h for 40 ℃; Shake a bottle rotating speed and be controlled at 150r/min, this nutrient solution is seeded to culture matrix.
5. method for preparing the 1-DNJ is characterized in that may further comprise the steps:
A. bacillus subtilis Py2 is carried out activation; Get glycerine and guarantee that medium component is 1% beef extract-peptone, 0.5% yeast extract in the inoculum concentration access activation medium of bacterial classification by 1% (v:/v) of Tibetan; 1% sodium chloride; Cultivate 16h for 40 ℃, shake a bottle rotating speed and be controlled at 150r/min, get seed culture fluid;
C. raw material is added water immersion 12-24h under 10-40 ℃ of 3 times of quality, place high-pressure sterilizing pot, in 121 ℃ of boiling 15-60min; After the cooling, the seed culture fluid that inserts 2% (v/m) ferments, and fermentation condition is: temperature 20-40 ℃; Humidity 70-95%, fermentation time are 12-96h;
D. the alpha-glucosidase restrainer in the bacillus subtilis Py2 fermentation soybean is purified: through activated carbon column chromatography, cation-exchange chromatography; Obtain the alpha-glucosidase restrainer monomer through preparation type TLC chromatography; Through mass spectrum and its molecular weight of nuclear magnetic resonance spectroscopy is 163Da, is the 1-DNJ.
Description of drawings
Fig. 1 is that bacillus subtilis (Bacillus Subtilis) Py2 ferments with the soybean of different cultivars, and it is active that the alpha-glucosidase of 40 ℃ of bottom fermentation 48h gained fermentation soybeans suppresses.Soybean varieties is respectively: 1, and section rich 6; 2, middle yellow 20; 3, close rich 35; 4, east farming 46; 5, Henan beans 28; 6, No. 8, middle beans; 7, No. 6, granule; 8, U.S.'s imported soybean.
Fig. 2 is that bacillus subtilis (Bacillus Subtilis) Py2 is a raw material with commercially available soybean, and fermentation different time, the alpha-glucosidase of fermentation soybean suppress active.
The mass spectrogram of the alpha-glucosidase restrainer that Fig. 3 is purified.
Biological material specimens preservation explanation
China Committee for Culture Collection of Microorganisms of depositary institution common micro-organisms center
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, address Institute of Microorganism, Academia Sinica
Preservation date on March 26th, 2012
Deposit number CGMCC NO.5934
Classification name bacillus subtilis Bacillus subtilis
The specific embodiment
The present invention is described further below in conjunction with instantiation, but is not restriction the present invention.
Step 1: the preparation of seed culture fluid: get glycerine and guarantee that medium component is 1% beef extract-peptone, 0.5% yeast extract, 1% sodium chloride in the inoculum concentration access activation medium of bacterial classification by 1% (v:/v) of Tibetan.Cultivate 16h for 40 ℃, shake a bottle rotating speed and be controlled at 150r/min, this nutrient solution is the seed culture fluid that is seeded to soybean;
The water that the commercially available soya bean 1Kg of step 2. adds 3 times of quality soaks 16h down at 20 ℃, places high-pressure sterilizing pot, and in 121 ℃ of boiling 40min, after the cooling, the seed culture fluid that inserts 2% (v/m) ferments;
Step 3: 40 ℃ of fermentation temperatures, humidity 90%, fermentation time is 48h, promptly obtains fermented soybean food.
Claims (8)
1. one kind has the fermented soybean food that alpha-glucosidase suppresses activity, and it is characterized in that: with soybean or other edible grain is raw material, is fermentation strain with bacillus subtilis Py2, forms through pure-blood ferment.
2. fermented soybean food as claimed in claim 1, it is characterized in that: the preparation method may further comprise the steps: soaking soybean, sterilization, inoculation, fermentation, fermentation condition is: temperature 20-40 ℃, humidity 70-95%, fermentation time are 12-96h.
3. fermented soybean food as claimed in claim 2; It is characterized in that: said immersion raw material and sterilization steps are: the water that soybean adds 3 times of quality soaks 12-24h down at 10-40 ℃; Place high-pressure sterilizing pot, obtain culture matrix in 121 ℃ of boiling 15-60min.
4. fermented soybean food as claimed in claim 2 is characterized in that: said inoculation step is: bacterial strain is carried out activation, get glycerine and guarantee in the inoculum concentration access activation medium of bacterial classification by 1% (v:/v) of Tibetan; Medium component is 1% beef extract-peptone; 0.5% yeast extract, 1% sodium chloride is cultivated 16h for 40 ℃; Shake a bottle rotating speed and be controlled at 150r/min, this nutrient solution is seeded to culture matrix.
5. the processed food that is prepared from each said fermented food of claim 1-5.
6. described fermented food of claim 1 or the described processed food of claim 5 are used for the purposes of function of blood sugar reduction food.
7. method for preparing the 1-DNJ is characterized in that may further comprise the steps:
A. bacillus subtilis Py2 is carried out activation; Get glycerine and guarantee that medium component is 1% beef extract-peptone, 0.5% yeast extract in the inoculum concentration access activation medium of bacterial classification by 1% (v:/v) of Tibetan; 1% sodium chloride; Cultivate 16h for 40 ℃, shake a bottle rotating speed and be controlled at 150r/min, get seed culture fluid;
C. raw material is added water immersion 12-24h under 10-40 ℃ of 3 times of quality, place high-pressure sterilizing pot, in 121 ℃ of boiling 15-60min; After the cooling, the seed culture fluid that inserts 2% (v/m) ferments, and fermentation condition is: temperature 20-40 ℃; Humidity 70-95%, fermentation time are 12-96h;
D. the alpha-glucosidase restrainer in the bacillus subtilis Py2 fermentation soybean is purified: through activated carbon column chromatography, cation-exchange chromatography; Obtain the alpha-glucosidase restrainer monomer through preparation type TLC chromatography; Through mass spectrum and its molecular weight of nuclear magnetic resonance spectroscopy is 163Da, is the 1-DNJ.
8. the 1-DNJ that is obtained by the said method of claim 7 is used as the purposes of function of blood sugar reduction.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105192592A (en) * | 2015-08-06 | 2015-12-30 | 武汉骏安生物科技有限公司 | Laba beans with function of reducing blood glucose and processing method thereof |
| CN105982035A (en) * | 2015-02-12 | 2016-10-05 | 上海市农业科学院 | Natto preparing method for increasing inhibition ratio of natto water extract to alpha-glucosidase |
| CN107373563A (en) * | 2017-08-22 | 2017-11-24 | 延边大学 | It is a kind of to facilitate Korean nationality's salty sauce powder and its eating method |
| CN115386514A (en) * | 2022-08-03 | 2022-11-25 | 武汉中博绿亚生物科技有限公司 | Bacillus subtilis (SCS-06) and application thereof |
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| JP2008263929A (en) * | 2007-04-25 | 2008-11-06 | Mitsukan Group Honsha:Kk | New bacillus natto and soft fermented soybeans produced by using the bacillus natto |
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| WO2011031020A2 (en) * | 2009-09-09 | 2011-03-17 | Cj Cheiljedang Corporation | Method for preparing a fermented soybean meal using bacillus strains |
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2012
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Patent Citations (4)
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105982035A (en) * | 2015-02-12 | 2016-10-05 | 上海市农业科学院 | Natto preparing method for increasing inhibition ratio of natto water extract to alpha-glucosidase |
| CN105982035B (en) * | 2015-02-12 | 2019-09-24 | 上海市农业科学院 | A kind of natto water extract that improves is to the natto preparation method of alpha-glucosaccharase enzyme inhibition rate |
| CN105192592A (en) * | 2015-08-06 | 2015-12-30 | 武汉骏安生物科技有限公司 | Laba beans with function of reducing blood glucose and processing method thereof |
| CN107373563A (en) * | 2017-08-22 | 2017-11-24 | 延边大学 | It is a kind of to facilitate Korean nationality's salty sauce powder and its eating method |
| CN115386514A (en) * | 2022-08-03 | 2022-11-25 | 武汉中博绿亚生物科技有限公司 | Bacillus subtilis (SCS-06) and application thereof |
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Application publication date: 20120829 |