CN101555275B - Method for preparing alpha-amylase inhibitor by enzymatic method - Google Patents
Method for preparing alpha-amylase inhibitor by enzymatic method Download PDFInfo
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- CN101555275B CN101555275B CN2009100393279A CN200910039327A CN101555275B CN 101555275 B CN101555275 B CN 101555275B CN 2009100393279 A CN2009100393279 A CN 2009100393279A CN 200910039327 A CN200910039327 A CN 200910039327A CN 101555275 B CN101555275 B CN 101555275B
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Abstract
The invention discloses a method for preparing an alpha-amylase inhibitor by an enzymatic method. In the method, white kidney beans are used as raw materials and are subjected to soaking, grinding andcentrifugating; supernatant fluid is processed by steps by complex enzyme and is subjected to ion-exchange chromatography and hyperfiltration to prepare the alpha-amylase inhibitor. In the method, th e white kidney beans are used as raw materials and the alpha-amylase inhibitor is prepared by the enzymatic method, therefore, the alpha-amylase inhibitor contained in the white kidney beans is effectively released to conduce to extraction and purification, thus reducing extraction cost. The method has mild reaction conditions, high purity of obtained products, good effect on inhibiting the amylase and inhibition activity as high as 70-95%, and can be widely applied in such fields as medicines, health-care food and the like.
Description
Technical field
The present invention relates to a kind of preparation method of amylase inhibitor.
Background technology
Present global obesity patient's quantity rises just with surprising rapidity, and scholarly forecast is with present development speed, and by 2230, all Americans will become the obese patient; And in Britain, in the period of nearly 20, overweight and obese person rises to 54% by 15% among the male sex, and the women then rises to 45%.Even if in the such developing country of China, a fat serious public health problem that has become China facing.China obese person is considerably beyond 9,000 ten thousand at present, and overweight person is up to 200,000,000.Scholarly forecast, Chinese obese people will surpass 200,000,000 in following 10 years.And obesity not only can cause many diseases such as diabetes, hypertension, cancer, also can make the people early death.There are data to show, 1.3～2 times of the dangerous right and wrong obese person that the obese person is early death.
Scientific research shows, obesity, lipomatosis, arteriosclerosis, hyperlipidemia, diabetes etc., often relevant with the intravital amylase activity of people, can block the starch transforming glucose by suppressing diastatic activity, thereby suppress the growing amount that blood sugar concentration raises and reduces fat.
The alpha-amylase inhibitor that the present invention relates to is a kind of glycoside hydrolase inhibitor, it is by suppressing the activity of interior saliva of enteron aisle and pancreatic amylase, hinder the hydrolysis and the digestion of carbohydrate in the food, optionally reduce sugar part picked-up, the content of lowering blood glucose and blood fat, reduce insulin secretion, reduce lipogenesis, thereby play the effect of hypoglycemic, fat-reducing and prevention of obesity.This kind alpha-amylase inhibitor derives from green plants, without any side effect, is a kind of glycoprotein to human body.Therefore, alpha-amylase inhibitor will have vast market prospect as the medicine or the healthcare products of a kind of prevention and treatment obesity and treatment diabetes.
Alpha-amylase inhibitor is a kind of glycoprotein, this glycoprotein can with the carbohydrate molecule forming composite, thereby the effect of blocking-up carbohydrate digestive ferment is so can effectively suppress the carbohydrate absorption.From plant seeds such as wheat, barley, jowar, cereal, peanut, soybean, taro, wild Amaranthus, separated obtaining alpha-amylase inhibitor at present, but the alpha-amylase inhibitor activity of extracting is higher from white waist beans.
Summary of the invention
The purpose of this invention is to provide a kind of method of utilizing biological enzyme to prepare alpha-amylase inhibitor, this method reaction conditions gentleness, the product purity height of gained, effective to diastatic inhibition.
Purpose of the present invention is achieved by taking following technical scheme:
A kind of method of utilizing biological enzyme to prepare alpha-amylase inhibitor is characterized in that with white waist beans be raw material, through soaking, pulverize, centrifugal, supernatant liquor prepares alpha-amylase inhibitor through ion exchange chromatography, ultrafiltration again with saccharifying enzyme and lipase step-by-step processing.Specifically may further comprise the steps:
(1) pre-treatment: white waist beans are soaked, and pulverize then, and the material that obtains carries out centrifugal treating;
(2) enzymic hydrolysis: the centrifugal supernatant liquor that goes out is adjusted pH value to 4.0～5.0, be warmed up to 45～50 ℃, add saccharifying enzyme reaction 1～12 hour, be warmed up to 60～70 ℃ of enzymes that go out, treat to adjust pH value to 6.0～7.0 after temperature drops to room temperature, be warmed up to 45～50 ℃ again, add lipase reaction 1～4 hour, be warmed up to 60～70 ℃ of enzymes that go out then;
(3) refining: the solution behind the enzyme that goes out again through the ultra-filtration membrane ultrafiltration, concentrates after cation exchange resin layer is analysed, and obtains alpha-amylase inhibitor after the drying.
The weight ratio of white waist beans and water is 1: 7～10 in the described step (1).
The time that white waist beans are soaked in the described step (1) is 4～24 hours.
The add-on of saccharifying enzyme is 0.1～1% of a white waist beans weight in the described step (2).
The add-on of lipase is 0.1～0.5% of a white waist beans weight in the described step (2).
Zeo-karb in the described step (3) is a strongly acidic styrene type cation exchange resin.
The flow velocity of solution by Zeo-karb that goes out in the described step (3) behind the enzyme is 1～2.5mL/min.
Ultra-filtration membrane in the described step (3) is the rolling ultra-filtration membrane.
Solution after chromatography is handled in the described step (3) is with relative velocity 1～2mL/min/cm
2By the rolling ultra-filtration membrane.
The present invention is a raw material with white waist beans, utilizes biological enzyme to prepare alpha-amylase inhibitor, and the alpha-amylase inhibitor that white waist beans are included discharges effectively, and favourable extraction and purifying reduce extraction cost.Reaction conditions gentleness of the present invention, the product purity height of gained, effective to diastatic inhibition, it suppresses active in 70%～95%, can be widely used in fields such as medicine and protective foods.
Embodiment
Below by specific embodiment the present invention is set forth, but protection scope of the present invention is not limited thereto.The person of an ordinary skill in the technical field all can realize purpose of the present invention according to above content disclosed by the invention.
Embodiment 1
Take by weighing white waist beans 100g, add entry, soaked 12 hours, pulverize according to 1: 10 ratio.The material that is obtained is centrifugal, and supernatant liquor is adjusted pH value to 4.5 ± 0.1, is warmed up to 45 ℃, and the saccharifying enzyme reaction of adding 0.2% 1 hour is warmed up to 70 ℃ of enzymes that go out.Treat to adjust pH value to 6.5 ± 0.1 after temperature drops to room temperature, be warmed up to 50 ℃ again, the lipase reaction of adding 0.1% 1 hour is warmed up to 70 ℃ of enzymes that go out then.Go out solution behind the enzyme with the flow velocity of 2mL/min by strongly acidic styrene type cation exchange resin, with the solution after the plastic resin treatment at 40 ℃ with relative velocity 1.5mL/min/cm
2By the rolling ultra-filtration membrane, the product behind the membrane filtration is through concentrating, and drying obtains the alpha-amylase inhibitor product.The inhibition activity of product is about 90%.
Embodiment 2
Take by weighing white waist beans 100g, add entry, soaked 24 hours, pulverize according to 1: 10 ratio.The material that is obtained is centrifugal, and supernatant liquor is adjusted pH value to 4.8 ± 0.1, is warmed up to 45 ℃, and the saccharifying enzyme reaction of adding 0.5% 4 hours is warmed up to 70 ℃ of enzymes that go out.Treat to adjust pH value to 6.3 ± 0.1 after temperature drops to room temperature, be warmed up to 50 ℃ again, the lipase reaction of adding 0.2% 1 hour is warmed up to 70 ℃ of enzymes that go out then.Go out solution behind the enzyme with the flow velocity of 2mL/min by strongly acidic styrene type cation exchange resin, with the solution after the plastic resin treatment at 40 ℃ with relative velocity 1.5mL/min/cm
2By the rolling ultra-filtration membrane, the product behind the membrane filtration is through concentrating, and drying obtains the alpha-amylase inhibitor product.The inhibition activity of product is about 87%.
Embodiment 3
Take by weighing white waist beans 100g, add entry, soaked 8 hours, pulverize according to 1: 7 ratio.The material that is obtained is centrifugal, and supernatant liquor is adjusted pH value to 4.3 ± 0.1, is warmed up to 45 ℃, and the saccharifying enzyme reaction of adding 0.1% 4 hours is warmed up to 60 ℃ of enzymes that go out.Treat to adjust pH value to 6.3 ± 0.1 after temperature drops to room temperature, be warmed up to 50 ℃ again, the lipase reaction of adding 0.1% 1 hour is warmed up to 60 ℃ of enzymes that go out then.Go out solution behind the enzyme with the flow velocity of 1mL/min by strongly acidic styrene type cation exchange resin, with the solution after the plastic resin treatment at 40 ℃ with relative velocity 1mL/min/cm
2By the rolling ultra-filtration membrane, the product behind the membrane filtration is through concentrating, and drying obtains the alpha-amylase inhibitor product.The inhibition activity of product is about 79%.
Embodiment 4
Take by weighing white waist beans 100g, add entry, soaked 12 hours, pulverize according to 1: 8 ratio.The material that is obtained is centrifugal, and supernatant liquor is adjusted pH value to 4.3 ± 0.1, is warmed up to 45 ℃, and the saccharifying enzyme reaction of adding 0.3% 4 hours is warmed up to 70 ℃ of enzymes that go out.Treat to adjust pH value to 6.3 ± 0.1 after temperature drops to room temperature, be warmed up to 50 ℃ again, the lipase reaction of adding 0.3% 1 hour is warmed up to 70 ℃ of enzymes that go out then.Go out solution behind the enzyme with the flow velocity of 1.5mL/min by strongly acidic styrene type cation exchange resin, with the solution after the plastic resin treatment at 40 ℃ with relative velocity 1.5mL/min/cm
2By the rolling ultra-filtration membrane, the product behind the membrane filtration is through concentrating, and drying obtains the alpha-amylase inhibitor product.The inhibition activity of product is about 83%.
Claims (10)
1. a method of utilizing biological enzyme to prepare alpha-amylase inhibitor is characterized in that, is raw material with white waist beans, through soaking, pulverize, centrifugal, supernatant liquor prepares alpha-amylase inhibitor through ion exchange chromatography, ultrafiltration again with saccharifying enzyme and lipase step-by-step processing.
2. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 1 is characterized in that, may further comprise the steps:
(1) pre-treatment: white waist beans are soaked, and pulverize then, and the material that obtains carries out centrifugal treating;
(2) enzymic hydrolysis: the centrifugal supernatant liquor that goes out is adjusted pH value to 4.0～5.0, be warmed up to 45～50 ℃, add saccharifying enzyme reaction 1～12 hour, 60～70 ℃ of enzymes that go out heat up, treat to adjust pH value to 6.0～7.0 after temperature drops to room temperature, be warmed up to 45～50 ℃ again, add lipase reaction 1～4 hour, be warmed up to 60～70 ℃ of enzymes that go out then;
(3) refining: the solution behind the enzyme that goes out again through the ultra-filtration membrane ultrafiltration, concentrates after cation exchange resin layer is analysed, and obtains alpha-amylase inhibitor after the drying.
3. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the weight ratio of white waist beans and water is 1: 7～10 in the described step (1).
4. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the time that white waist beans are soaked in the described step (1) is 4～24 hours.
5. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the add-on of saccharifying enzyme is 0.1～1% of a white waist beans weight in the described step (2).
6. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the add-on of lipase is 0.1～0.5% of a white waist beans weight in the described step (2).
7. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the Zeo-karb in the described step (3) is a strongly acidic styrene type cation exchange resin.
8. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the solution behind the enzyme that goes out in the described step (3) is 1～2.5mL/min by the flow velocity of Zeo-karb.
9. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the ultra-filtration membrane in the described step (3) is the rolling ultra-filtration membrane.
10. the method for utilizing biological enzyme to prepare alpha-amylase inhibitor according to claim 2 is characterized in that, the solution after chromatography is handled in the described step (3) is with relative velocity 1～2mL/min/cm
2By the rolling ultra-filtration membrane.
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| CN103638089B (en) * | 2013-11-29 | 2016-01-27 | 广东省食品工业研究所 | A kind of method utilizing microwave-compound enzyme coupled method to extract alpha-amylase inhibitor from Semen Phaseoli Vulgaris |
| CN104131057B (en) * | 2014-07-23 | 2017-01-18 | 山西大学 | Functional white kidney bean polypeptide and preparation method and application thereof |
| CN105747231B (en) * | 2016-03-16 | 2018-12-04 | 宝健（北京）生物技术有限公司 | The extraction and preparation technique of starch Inhibitors in a kind of navy bean |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1665521A (en) * | 2002-06-28 | 2005-09-07 | 法蒙凯姆实验室公司 | Purified amylase inhibitor and novel process for obtaining the same |
| WO2007053846A1 (en) * | 2005-11-01 | 2007-05-10 | Soft Gel Technologies, Inc. | Compositions of hoodia gordonii and pinolenic acid derivatives |
| CN101340923A (en) * | 2005-12-22 | 2009-01-07 | 因德纳有限公司 | Ethanol-precipitated phaseolus vulgaris extracts, their use and formulations |
| CN101366439A (en) * | 2008-08-30 | 2009-02-18 | 山东阜丰生物科技开发有限公司 | Method for improving extracting yield of soy protein |
-
2009
- 2009-05-08 CN CN2009100393279A patent/CN101555275B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1665521A (en) * | 2002-06-28 | 2005-09-07 | 法蒙凯姆实验室公司 | Purified amylase inhibitor and novel process for obtaining the same |
| WO2007053846A1 (en) * | 2005-11-01 | 2007-05-10 | Soft Gel Technologies, Inc. | Compositions of hoodia gordonii and pinolenic acid derivatives |
| CN101340923A (en) * | 2005-12-22 | 2009-01-07 | 因德纳有限公司 | Ethanol-precipitated phaseolus vulgaris extracts, their use and formulations |
| CN101366439A (en) * | 2008-08-30 | 2009-02-18 | 山东阜丰生物科技开发有限公司 | Method for improving extracting yield of soy protein |
Non-Patent Citations (8)
| Title |
|---|
| J.J Marshall,C.M Lauda.Purification and properties of phaseolamin, an inhibitor of alpha-amylase, from the kidney bean, Phaseolus vulgaris.《The Journal of Biological Chemistry》.1975,第250卷(第20期),8030-8031. * |
| J.JMarshall C.M Lauda.Purification and properties of phaseolamin |
| 张晓琦等.白豆α-淀粉酶抑制剂糖蛋白的提取纯化及降血糖活性研究.《药物生物技术》.2007,第14卷(第6期),406-410. * |
| 张晓琦等.白豆α-淀粉酶抑制剂糖蛋白的提取纯化及降血糖活性研究.<<药物生物技术>>.2007,第14卷(第6期),406-410. |
| 杨秀芳等.白豆中α-淀粉酶抑制剂的提取及其酶学性质的研究.《应用化工》.2007,第36卷(第8期),755-757. * |
| 杨秀芳等.白豆中α-淀粉酶抑制剂的提取及其酶学性质的研究.<<应用化工>>.2007,第36卷(第8期),755-757. |
| 赵蓉等.白芸豆中α-淀粉酶抑制剂的研究.《中成药》.2008,第30卷(第9期),1355-1357. * |
| 赵蓉等.白芸豆中α-淀粉酶抑制剂的研究.<<中成药>>.2008,第30卷(第9期),1355-1357. |
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