CN102226212A - Method for preparing amylose having narrow molecular weight distribution range - Google Patents
Method for preparing amylose having narrow molecular weight distribution range Download PDFInfo
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- CN102226212A CN102226212A CN2011101202863A CN201110120286A CN102226212A CN 102226212 A CN102226212 A CN 102226212A CN 2011101202863 A CN2011101202863 A CN 2011101202863A CN 201110120286 A CN201110120286 A CN 201110120286A CN 102226212 A CN102226212 A CN 102226212A
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Abstract
The invention discloses a method for preparing amylose having a narrow molecular weight distribution range, and aims to provide a simple and low-cost preparation method for separating amylose and amylopectin from retrogradation resistant starch based on the characteristic that only amylose and amylopectin within a certain molecular weight range participate in retrogradation under specific conditions in the starch retrogradation process. The method comprises the following steps: carrying out enzymolysis on the retrogradation resistant starch by thermostable amylase, thus obtaining purified retrogradation resistant starch; dissolving the purified retrogradation resistant starch in 2-4mol/L alkaline liquid; and carrying out precipitation separation on the retrogradation resistant starch dissolved in the alkaline liquid by n-butyl alcohol to obtain the amylose, and carrying out precipitation separation by ethanol to obtain the amylopectin. The method disclosed by the invention is simple, is low in cost, is short in separation time and can obtain a large number of separation products.
Description
Technical field
The present invention relates to the method that a kind of legal system of bringing back to life is equipped with the narrower amylose starch of range of molecular weight distributions.
Background technology
Resistant starch has prevention, alleviates and physiological functions such as metabolism, the promotion mineral substance of treatment constipation, control of diabetes, promotion lipoid and cholesterol absorb, enhancing disease resistance.European resistant starch association had carried out the notion of Resistant starch (RS) to define and standard in 1996, promptly not by the general name of the starch of the little intestinal absorption of healthy human body and hydrolyzate thereof, the Resistant starch of bringing back to life is meant after the gelatinization starch of cool aging again, its essence is to have formed crystallization between the amylose starch, this crystallization can not be by little intestinal digestion in the normal human, but can part be utilized by microorganism in large intestine.
Gelatinization and the preparation of organic solvent deposit method are adopted in the preparation of amylose starch and amylopectin usually in the starch, the starch pasting process makes the starch ball fractured, amylose starch wherein and amylopectin are free to form starch solution in water, then that water-insoluble is centrifugal or remove by filter, the water solubles adopts isolates amylose starch to the method for wherein adding propyl carbinol, and surplus solution adds ethanol etc. again and separates wherein amylopectin.The range of molecular weight distributions broad of amylopectin that aforesaid method obtains and amylose starch, at the research starch property, in experimentations such as viscosity, degree of melting, degree of bringing back to life, the too wide experimental result circulation ratio that causes of straight chain and amylopectin range of molecular weight distributions is not good, different researchists are not to having comparability between the same starch property result of study, for straight chain and the amylopectin that obtains the certain molecular weight scope, needs employing gel chromatography etc. separates, cost an arm and a leg, disengaging time is long, and separated product is few.
Summary of the invention
The present invention is in order to overcome weak point of the prior art, provide a kind of straight chain that the certain molecular weight scope is only arranged under the specified conditions in the starch retrogradation process and amylopectin of utilizing to participate in the characteristic of bringing back to life, from the Resistant starch of bringing back to life, separate amylose starch, can also obtain amylopectin, the preparation method is simple, the method for the amylose starch that the preparation range of molecular weight distributions that cost is low is narrower.
The present invention is achieved through the following technical solutions:
A kind of legal system of bringing back to life is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that: comprise the steps:
(1) Resistant starch of will bringing back to life uses the alpha-amylase enzymolysis, obtains the purifying Resistant starch of bringing back to life; The condition of enzymolysis is: diastatic add-on is the 1200U/100g Resistant starch of bringing back to life, and hydrolysis temperature is 90-100 ℃, and enzymolysis time is 10-30min, adopt then 3000 rev/mins centrifugal, the Resistant starch of bringing back to life of the centrifugal acquisition purifying in precipitation washing back after centrifugal;
(2) purifying that step (1) the is obtained Resistant starch concentration of bringing back to life is the alkali lye dissolving of 2-4mol/L;
(3) will add propyl carbinol in the Resistant starch of bringing back to life after the alkali lye dissolving, the centrifugal afterwards amylose starch precipitation that obtains obtains the narrower amylose starch of molecular weight ranges after the lyophilize of amylose starch precipitation.
As required, surplus solution can further separate and obtains amylopectin.To add dehydrated alcohol in the rest solution after step (3) the precipitation amylose starch, the centrifugal afterwards amylopectin precipitation that obtains obtains the narrower amylopectin of molecular weight ranges after the lyophilize of amylopectin precipitation.
Described alkali lye is sodium hydroxide or potassium hydroxide aqueous solution.
The raw material for preparing the Resistant starch of bringing back to life is any in sweet potato starch, yam starch, W-Gum, wheat starch, barley starch, the legume starch.
Amylose starch precipitation lyophilize temperature is-65 ℃, and the time is 3 days.
Amylopectin precipitation lyophilize temperature is-65 ℃, and the time is 3 days.
The present invention has following technique effect:
Method of the present invention utilizes in the starch retrogradation process polymerization degree only to participate in the characteristic of bringing back to life for the amylose starch of 20-100 and amylopectin, from the Resistant starch of bringing back to life, separate amylose starch, also can separate obtaining amylopectin as required, obtain narrower amylose starch of range of molecular weight distributions and amylopectin.Preparation method of the present invention is simple, and cost is low, and disengaging time is short, and the separated product that obtains is many, and the general fractional dose of gel chromatography is in the gram level, and the inventive method can be in feather weight.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
(1) sweet potato starch 100 gram and water being mixed and made into concentration is 10% starch milk, put into pressure kettle behind 90 ℃ of gelatinization 30min and carry out autoclaving, the high pressure temperature is 105-120 ℃, the high pressure time is 30min, wear out after the taking-up and obtain the semi-solid Resistant starch of bringing back to life, aging temperature is 15 ℃, and digestion time is 6h.
(2) semi-solid that step (1) the is obtained Resistant starch of bringing back to life adopts the alpha-amylase enzymolysis, enzymatic hydrolysis condition is: the semi-solid that every 100g step (1) the obtains Resistant starch of bringing back to life adds amylase 1200U, hydrolysis temperature is 90-100 ℃, enzymolysis time is 10min, adopt then 3000 rev/mins centrifugal, precipitation is with the triploid ponding centrifugal acquisition purifying Resistant starch of bringing back to life of giving a baby a bath on the third day after its birth time afterwards.
(3) purifying that obtains behind the enzymolysis is brought back to life to adopt concentration be the aqueous sodium hydroxide solution dissolving of 2mol/L to Resistant starch.Add the propyl carbinol of 2 times of volumes of alkali lye in the Resistant starch of bringing back to life after the alkali lye dissolving, afterwards, 3000 rev/mins of centrifugal amylose starch precipitations that obtain are outwelled the upper strata propyl carbinol, the dehydrated alcohol that adds 1 times of volume in the rest solution, the centrifugal afterwards amylopectin precipitation that obtains.-65 ℃ of lyophilizes of amylose starch precipitation obtain the narrower amylose starch of molecular weight ranges 6.5 grams after 3 days, the polymerization degree of the amylose starch that obtains is 65-75.-65 ℃ of lyophilizes of amylopectin precipitation obtain the narrower amylopectin of molecular weight ranges 4.7 grams after 3 days, the polymerization degree of amylopectin is 140-260.
The sweet potato starch of same sample adopts gelatinization and the organic solvent deposit method in the background technology, the amylose molecule weight range that is separated to is 12-897 for the starch polymerization degree, the amylopectin molecular weight ranges is 38-2890 for the starch polymerization degree, through amylose starch that method of the present invention is separated to and amylopectin molecular weight distribution than ative starch direct isolating amylose starch and amylopectin range of molecular weight distributions narrow more than 5 times.
Embodiment 2
(1) W-Gum 100 gram and water being mixed and made into concentration is 10% starch milk, put into pressure kettle behind 90 ℃ of gelatinization 30min and carry out autoclaving, the high pressure temperature is 105-120 ℃, the high pressure time is 30min, wear out after the taking-up and obtain the semi-solid Resistant starch of bringing back to life, aging temperature is 15 ℃, and digestion time is 6h.
(2) Resistant starch of bringing back to life that step (1) is obtained adopts the alpha-amylase enzymolysis, enzymatic hydrolysis condition is: the semi-solid that every 100g step (1) the obtains Resistant starch of bringing back to life adds amylase 1200U, hydrolysis temperature is 90-100 ℃, enzymolysis time is 10min, adopt then 3000 rev/mins centrifugal, precipitation is with the triploid ponding centrifugal acquisition purifying Resistant starch of bringing back to life of giving a baby a bath on the third day after its birth time afterwards.
(3) purifying that obtains behind the enzymolysis is brought back to life to adopt concentration be the aqueous sodium hydroxide solution dissolving of 2mol/L to Resistant starch.Add the propyl carbinol of 2 times of volumes of alkali lye in the Resistant starch of bringing back to life after the alkali lye dissolving, afterwards, 3000 rev/mins of centrifugal amylose starch precipitations that obtain are outwelled the upper strata propyl carbinol, the dehydrated alcohol that adds 1 times of volume in the rest solution, the centrifugal afterwards amylopectin precipitation that obtains.-65 ℃ of lyophilizes of amylose starch precipitation obtain the narrower amylose starch of molecular weight ranges 6.9 grams after 3 days ,-65 ℃ of lyophilizes of amylopectin precipitation obtain the narrower amylopectin of molecular weight ranges 3.7 grams after 3 days.The polymerization degree of the amylose starch for preparing is 43-87, and the amylopectin polymerization degree is 110-220.
The W-Gum of same sample adopts gelatinization and the organic solvent deposit method in the background technology, separating the amylose molecule weight range that obtains is 14-634 for the starch polymerization degree, the amylopectin molecular weight ranges is 22-2456 for the starch polymerization degree, through amylose starch that method of the present invention is separated to and amylopectin molecular weight distribution than ative starch direct isolating amylose starch and amylopectin range of molecular weight distributions narrow more than 5 times.
Embodiment 3
(1) yam starch 100 gram and water being mixed and made into concentration is 10% starch milk, put into pressure kettle behind 90 ℃ of gelatinization 30min and carry out autoclaving, the high pressure temperature is 105-120 ℃, the high pressure time is 30min, wear out after the taking-up and obtain the semi-solid Resistant starch of bringing back to life, aging temperature is 15 ℃, and digestion time is 6h.
(2) Resistant starch of bringing back to life that step (1) is obtained adopts the alpha-amylase enzymolysis, enzymatic hydrolysis condition is: the semi-solid that every 100g step (1) the obtains Resistant starch of bringing back to life adds amylase 1200U, hydrolysis temperature is 90-100 ℃, enzymolysis time is 10min, adopt then 3000 rev/mins centrifugal, precipitation is with the triploid ponding centrifugal acquisition purifying Resistant starch of bringing back to life of giving a baby a bath on the third day after its birth time afterwards.
(3) purifying that obtains behind the enzymolysis is brought back to life to adopt concentration be the aqueous sodium hydroxide solution dissolving of 2mol/L to Resistant starch.Add the propyl carbinol of 2 times of volumes of alkali lye in the Resistant starch of bringing back to life after the alkali lye dissolving, afterwards, 3000 rev/mins of centrifugal amylose starch precipitations that obtain are outwelled the upper strata propyl carbinol, the dehydrated alcohol that adds 1 times of volume in the rest solution, the centrifugal afterwards amylopectin precipitation that obtains.-65 ℃ of lyophilizes of amylose starch precipitation obtain the narrower amylose starch of molecular weight ranges 2.5 grams after 3 days ,-65 ℃ of lyophilizes of amylopectin precipitation obtain the narrower amylopectin of molecular weight ranges 1.7 grams after 3 days.The polymerization degree of the amylose starch for preparing is 135-189, and the polymerization degree of amylopectin is 110-340.
The sweet potato starch of same sample adopts gelatinization and the organic solvent deposit method in the background technology, separating the amylose molecule weight range that obtains is 18-1460 for the starch polymerization degree, the amylopectin molecular weight ranges is 68-3782 for the starch polymerization degree, through amylose starch that method of the present invention is separated to and amylopectin molecular weight distribution than ative starch direct isolating amylose starch and amylopectin range of molecular weight distributions narrow more than 5 times.
Claims (6)
1. the legal system of bringing back to life is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that: comprise the steps:
(1) Resistant starch of will bringing back to life uses the alpha-amylase enzymolysis, obtains the purifying Resistant starch of bringing back to life; The condition of enzymolysis is: diastatic add-on is the 1200U/100g Resistant starch of bringing back to life, and hydrolysis temperature is 90-100 ℃, and enzymolysis time is 10-30min, and is centrifugal then, the centrifugal again acquisition purifying Resistant starch of bringing back to life after the precipitation washing after centrifugal;
(2) purifying that step (1) the is obtained Resistant starch concentration of bringing back to life is the alkali lye dissolving of 2-4mol/L;
(3) will add propyl carbinol in the Resistant starch of bringing back to life after the alkali lye dissolving, the centrifugal afterwards amylose starch precipitation that obtains obtains the narrower amylose starch of molecular weight ranges after the lyophilize of amylose starch precipitation.
2. the legal system of bringing back to life according to claim 1 is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that: will add dehydrated alcohol in the rest solution after step (3) the precipitation amylose starch, the centrifugal afterwards amylopectin precipitation that obtains obtains the narrower amylopectin of molecular weight ranges after the lyophilize of amylopectin precipitation.
3. the legal system of bringing back to life according to claim 1 and 2 is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that, described alkali lye is sodium hydroxide or potassium hydroxide aqueous solution.
4. the legal system of bringing back to life according to claim 1 and 2 is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that the raw material for preparing the Resistant starch of bringing back to life is any in sweet potato starch, yam starch, W-Gum, wheat starch, barley starch, the legume starch.
5. the legal system of bringing back to life according to claim 1 is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that, amylose starch precipitation lyophilize temperature is-65 ℃, and the time is 3 days.
6. the legal system of bringing back to life according to claim 2 is equipped with the method for the narrower amylose starch of range of molecular weight distributions, it is characterized in that, amylopectin precipitation lyophilize temperature is-65 ℃, and the time is 3 days.
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| CN 201110120286 CN102226212B (en) | 2011-05-11 | 2011-05-11 | Method for preparing amylose having narrow molecular weight distribution range |
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| CN 201110120286 CN102226212B (en) | 2011-05-11 | 2011-05-11 | Method for preparing amylose having narrow molecular weight distribution range |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104926949A (en) * | 2015-07-10 | 2015-09-23 | 华东理工大学 | Preparation method for high purity amylose |
| CN106770395A (en) * | 2016-03-29 | 2017-05-31 | 天津商业大学 | The dynamic (dynamical) method of starch retrogradation is studied using peak area is selected |
| CN111848825A (en) * | 2020-07-17 | 2020-10-30 | 天津商业大学 | Method for preparing retrogradation-resistant amylose by using salt eutectic method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3881991A (en) * | 1969-01-24 | 1975-05-06 | Hayashibara Co | Process for producing amylose powders having a mean degree of polymerization between 20{14 30 |
-
2011
- 2011-05-11 CN CN 201110120286 patent/CN102226212B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3881991A (en) * | 1969-01-24 | 1975-05-06 | Hayashibara Co | Process for producing amylose powders having a mean degree of polymerization between 20{14 30 |
Non-Patent Citations (3)
| Title |
|---|
| 《粮食与油脂》 20091231 连喜军 等 alpha-淀粉酶水解马铃薯淀粉制备抗性淀粉 11-14 1-6 , * |
| XIJUN LIAN ET AL.: "A photographic approach to the possible mechanism of retrogradation of sweet potato starch.", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》, 20 October 2010 (2010-10-20), pages 125 - 128 * |
| 连喜军 等: "α-淀粉酶水解马铃薯淀粉制备抗性淀粉", 《粮食与油脂》, 31 December 2009 (2009-12-31), pages 11 - 14 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104926949A (en) * | 2015-07-10 | 2015-09-23 | 华东理工大学 | Preparation method for high purity amylose |
| CN106770395A (en) * | 2016-03-29 | 2017-05-31 | 天津商业大学 | The dynamic (dynamical) method of starch retrogradation is studied using peak area is selected |
| CN111848825A (en) * | 2020-07-17 | 2020-10-30 | 天津商业大学 | Method for preparing retrogradation-resistant amylose by using salt eutectic method |
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| CN102226212B (en) | 2013-09-25 |
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