CN104770737A - Phaseolus vulgaris alpha-amylase inhibitor extraction and separation method - Google Patents
Phaseolus vulgaris alpha-amylase inhibitor extraction and separation method Download PDFInfo
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- CN104770737A CN104770737A CN201510135921.3A CN201510135921A CN104770737A CN 104770737 A CN104770737 A CN 104770737A CN 201510135921 A CN201510135921 A CN 201510135921A CN 104770737 A CN104770737 A CN 104770737A
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Abstract
The present invention provides a phaseolus vulgaris alpha-amylase inhibitor extraction separation method, and relates to the field of food science biological active component extraction and separation, wherein the obtaining rate can be increased, and the yield can be improved. The method comprises: crushing phaseolus vulgaris to obtain phaseolus vulgaris crude powder, carrying out stirring extraction on the phaseolus vulgaris crude powder 3 times with water at a room temperature, filtering to obtain a filtrate, carrying out pressure reducing concentration on the filtrate to obtain a concentrated solution, carrying out boiling sterilization on the concentrated solution, adding a glucono delta-lactone coagulant, precipitating, filtering to obtain a precipitate, and sequentially carrying out decolorizing, dewatering, filtration draining, vacuum drying, and crushing so as to obtain the phaseolus vulgaris alpha-amylase inhibitor product.
Description
Technical field
The present invention relates to Food Science bioactive ingredients extraction and isolation field, particularly relate to the extraction separation method of navy bean alpha-amylase inhibitor.
Background technology
Navy bean a-amylase inhibitor extracts to form from pulse family kidney bean platymiscium navy bean (phaseoluscoccineus L., P.multiflorus willd.).Navy bean biolvgical name is scarlet runner bean, originates in Mexico and the A Genyan in America, has adapted to by artificial cultivation domestication the moist area, plateau that cools.The country that cultivated area is larger is the Britain in Argentina in America, the U.S., Mexico, Europe, the China, Japan etc. in Asia.All there is plantation each provinces and regions of present China, and the province that area is larger is Yunnan, Guizhou, Sichuan etc.
Navy bean a-amylase inhibitor is a kind of native starch blocking agent, and it can before the Starch Conversion of absorption is glucose by the a-amylase in human body, suppress it active, thus effectively blocks human body and absorb the heat produced from starch food.Amylase inhibitor be first through the effective Starch blockers of clinical verification, and have passed the certification of U.S. FDA (food and medicine Surveillance Authority) and U.S. GRAS (it is generally acknowledged safety), the food uniquely can expressing the function such as " absorption and digestion that can reduce starch in meals ", " coordinate rational diet and motion, can help to control body weight " on the packaging.Be mainly used in treatment and prevent and treat obesity, diabetes, also having the functions such as spleen-invigorating and kidney-strengthening.
At present, following flow process is mainly adopted for the extraction and isolation of navy bean alpha-amylase inhibitor both at home and abroad: raw material-pulverizing-water logging-centrifugation-supernatant-saltout (precipitation is dissolved again)-heat enzyme-centrifugation-supernatant-dialysis of going out-concentrated-drying-finished product.
These methods are all the production as navy bean alpha-amylase inhibitor higher degree mostly, and feature is that purity is high, but yield is extremely low.Most conduct uses in scientific research or costly medicine, and cannot use because of its high expensive in food, health products, thus there are many fake products to flood market in the market, therefore market are badly in need of the navy bean alpha-amylase inhibitor product used in a kind of applicable food, health products production.
Summary of the invention
Embodiments of the invention provide a kind of extraction separation method of navy bean alpha-amylase inhibitor, can improve pick-up rate, increase yield.
For achieving the above object, embodiments of the invention adopt following technical scheme:
An extraction separation method for navy bean alpha-amylase inhibitor, comprising:
Navy bean is pulverized, obtains navy bean meal; Described navy bean meal to be blunged at normal temperatures extraction 3 times, filter and obtain filtrate, described filtrate reduced in volume is become concentrate, described concentrate is boiled virus killing, add gluconolactone coagulating agent, precipitation, filter, obtain sediment; Decoloured by described sediment, dehydration, filtration is drained, vacuum drying, pulverizes, obtains navy bean alpha-amylase inhibitor product.
Preferably, described method specifically comprises:
S1, navy bean to be pulverized, cross 10 mesh sieves, obtain navy bean meal;
S2, modulate with sodium acid carbonate the running water that pH is 7.5;
S3, in described navy bean meal, add 10-15 described pH be doubly the running water of 7.5, stirs 4-6h, filter at 20-35 DEG C, collect first time filtrate and first time residue;
S4, being the running water of 7.5 adding 8-12 described pH doubly in described first time residue, at 20-35 DEG C, stirring 2-4h, filter, collecting second time filtrate and second time residue;
S5, in described second time residue, add 8-12 described pH be doubly the running water of 7.5, stirs 2-4h, filter, collect third time filtrate at 20-35 DEG C;
S6, merge described first time filtrate, second time filtrate and third time filtrate be mixing filtrate, described mixing filtrate is evaporated to the 2-4 of material quantity doubly at 65 DEG C, obtains concentrate;
S7,5-10min is boiled in the heating of described concentrate, add the gluconolactone of concentrated liquid measure 1%-2% while hot, fully dissolve, at 80 DEG C, be incubated 30-60min, then leave standstill more than 5h, protein is fully solidified, then centrifugal or filter, obtain sediment;
S8, described sediment is first washed one time with 75% alcohol, decolouring, then wash one time with 95% alcohol, dehydration, centrifugal or filter and drain, in 55-60 DEG C of vacuum drying, pulverizing, obtains navy bean alpha-amylase inhibitor product.
Beneficial effect of the present invention:
(1) the present invention adopts low temperature to extract, and does not namely cause protein denaturation, makes alpha-amylase inhibitor inactivation, does not cause starch gelatinization again, affects solution and filters.
(2) the present invention adopts sodium bicarbonate aqueous solution to adjust pH to be that the running water of 7.5 is to extract alpha-amylase inhibitor, he contributes to the stripping of navy bean protein and the stable of colloidal solution, the hydrolysis of CKIs matter, improves the solidification rate of protein in concentrate, increase yield.
(3) the present invention using gluconolactone as the agent of navy bean protein coagulating, change the method for alcohol precipitation, acid precipitation or the ammonium salt precipitation that protein precipitation is commonly used, changing alcohol precipitation process can make polysaccharide with protein coprecipitation out, make product sugar content too high, do not utilize product in treatment and prevent and treat obesity, the application in diabetes; After changing ammonium salt precipitation, the necessary fully spendable drawback in desalination side, makes navy bean proteins precipitate (navy bean alpha-amylase inhibitor) very safe, very agreeable to the taste using in food, health products.
(4) the present invention adopts concentrate to boil 5-10min, its object makes phytolectin inactivation exactly, navy bean alpha-amylase inhibitor can be used safely in food, health products, and can not damage because phytolectin has the existence of stronger toxic and side effect to people and mammal.
(5) the present invention adopts sediment first to wash one time with 75% alcohol, and decolouring, is washing one time with 95% alcohol, and dehydration, to ensure that product has more shallow color and luster, is convenient to use in food, health products.
Detailed description of the invention
Be clearly and completely described the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Embodiment 1:
Navy bean is pulverized, cross 10 mesh sieves, take navy bean meal 50kg, add with sodium bicarbonate aqueous solution adjust pH be 7.5 10 times, running water, stir at 20 DEG C and extract 4h, filter, collect first time filtrate, add in first time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 8 times, running water, stir at 20 DEG C and extract 2h, filter, collect second time filtrate, add in second time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 8 times, running water, stir at 20 DEG C and extract 2h, filter, collect third time filtrate, merge three filtrates and become mixing filtrate, mixing filtrate reduced pressure concentration at 65 DEG C becomes concentrate, described concentrate is 3 times of described mixing filtrate material quantity, 5-10min is boiled in described concentrate heating, add the gluconolactone of concentrated liquid measure 1% while hot, abundant dissolving, 30min is incubated at 80 DEG C, leave standstill more than 5h again, make the abundant coagulating agent of protein, centrifugal or filtration obtains sediment, sediment first washes one time with 75% alcohol, decolouring, one time is washed again with 95% alcohol, dehydration, centrifugal or filtration is drained, in 55-60 DEG C of vacuum drying, pulverize, obtain navy bean alpha-amylase inhibitor product 5.85kg.
Embodiment 2:
Navy bean is pulverized, cross 10 mesh sieves, take navy bean meal 50kg, add with sodium bicarbonate aqueous solution adjust pH be 7.5 15 times, running water, stir at 20 DEG C and extract 6h, filter, collect first time filtrate, add in first time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 12 times, running water, stir at 20 DEG C and extract 4h, filter, collect second time filtrate, add in second time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 12 times, running water, stir at 20 DEG C and extract 4h, filter, collect third time filtrate, merge three filtrates and become mixing filtrate, mixing filtrate reduced pressure concentration at 65 DEG C becomes concentrate, described concentrate is 3 times of described mixing filtrate material quantity, 5-10min is boiled in described concentrate heating, add the gluconolactone of concentrated liquid measure 1% while hot, abundant dissolving, 30min is incubated at 80 DEG C, leave standstill more than 5h again, make the abundant coagulating agent of protein, centrifugal or filtration obtains sediment, sediment first washes one time with 75% alcohol, decolouring, one time is washed again with 95% alcohol, dehydration, centrifugal or filtration is drained, in 55-60 DEG C of vacuum drying, pulverize, obtain navy bean alpha-amylase inhibitor product 6.12kg.
Embodiment 3:
Navy bean is pulverized, cross 10 mesh sieves, take navy bean meal 50kg, add with sodium bicarbonate aqueous solution adjust pH be 7.5 15 times, running water, stir at 35 DEG C and extract 6h, filter, collect first time filtrate, add in first time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 12 times, running water, stir at 35 DEG C and extract 4h, filter, collect second time filtrate, add in second time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 12 times, running water, stir at 35 DEG C and extract 4h, filter, collect third time filtrate, merge three filtrates and become mixing filtrate, mixing filtrate reduced pressure concentration at 65 DEG C becomes concentrate, described concentrate is 3 times of described mixing filtrate material quantity, 5-10min is boiled in described concentrate heating, add the gluconolactone of concentrated liquid measure 1% while hot, abundant dissolving, 30min is incubated at 80 DEG C, leave standstill more than 5h again, make the abundant coagulating agent of protein, centrifugal or filtration obtains sediment, sediment first washes one time with 75% alcohol, decolouring, one time is washed again with 95% alcohol, dehydration, centrifugal or filtration is drained, in 55-60 DEG C of vacuum drying, pulverize, obtain navy bean alpha-amylase inhibitor product 6.14kg.
Embodiment 4:
Navy bean is pulverized, cross 10 mesh sieves, take navy bean meal 50k8, add with sodium bicarbonate aqueous solution adjust pH be 7.5 15 times, running water, stir at 20 DEG C and extract 6h, filter, collect first time filtrate, add in first time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 12 times, running water, stir at 20 DEG C and extract 4h, filter, collect second time filtrate, add in second time residue with sodium bicarbonate aqueous solution adjust pH be 7.5 12 times, running water, stir at 20 DEG C and extract 4h, filter, collect third time filtrate, merge three filtrates and become mixing filtrate, mixing filtrate reduced pressure concentration at 65 DEG C becomes concentrate, described concentrate is 4 times of described mixing filtrate material quantity, 5-10min is boiled in described concentrate heating, add the gluconolactone of concentrated liquid measure 2% while hot, abundant dissolving, 30min is incubated at 80 DEG C, leave standstill more than 5h again, make the abundant coagulating agent of protein, centrifugal or filtration obtains sediment, sediment first washes one time with 75% alcohol, decolouring, one time is washed again with 95% alcohol, dehydration, centrifugal or filtration is drained, in 55-60 DEG C of vacuum drying, pulverize, obtain navy bean alpha-amylase inhibitor product 6.08kg.
Adopt method of the present invention, the pick-up rate of navy bean alpha-amylase inhibitor is: 11%-13%, and in prior art, the pick-up rate of navy bean alpha-amylase inhibitor is: 4%-6%.The pick-up rate of method navy bean alpha-amylase inhibitor that the present invention adopts is greatly improved, and adds the output of navy bean alpha-amylase inhibitor, reduces its cost in food, health products.
In the embodiment of the present invention, gluconolactone is applied in the production of navy bean alpha-amylase inhibitor as the agent of navy bean protein coagulating, navy bean alpha-amylase inhibitor product is used in food, health products very safe, very agreeable to the taste, and recovery rate is high, meet enterprise under the prerequisite of not faking, obtain good economic benefit.Gluconolactone use amount is the 1-2% of concentrated liquid measure.
In addition, concentrate is adopted to boil 5-10min in the invention process, though simple boils 5-10min, but make phytolectin inactivation in navy bean extract, navy bean alpha-amylase inhibitor can be used safely in food, health products, and can not damage because phytolectin has the existence of stronger toxic and side effect to people and mammal.
The above; be only the specific embodiment of the present invention, but protection scope of the present invention is not limited thereto, is anyly familiar with those skilled in the art in the technical scope that the present invention discloses; change can be expected easily or replace, all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should described be as the criterion with the protection domain of claim.
Claims (2)
1. an extraction separation method for navy bean alpha-amylase inhibitor, is characterized in that, comprising:
Navy bean is pulverized, obtains navy bean meal; Described navy bean meal to be blunged at normal temperatures extraction 3 times, filter and obtain filtrate, described filtrate reduced in volume is become concentrate, described concentrate is boiled virus killing, add gluconolactone coagulating agent, precipitation, filter, obtain sediment; Decoloured by described sediment, dehydration, filtration is drained, vacuum drying, pulverizes, obtains navy bean alpha-amylase inhibitor product.
2. method according to claim 1, is characterized in that, described method specifically comprises:
S1, navy bean to be pulverized, cross 10 mesh sieves, obtain navy bean meal;
S2, modulate with sodium acid carbonate the running water that pH is 7.5;
S3, in described navy bean meal, add 10-15 described pH be doubly the running water of 7.5, stirs 4-6h, filter at 20-35 DEG C, collect first time filtrate and first time residue;
S4, being the running water of 7.5 adding 8-12 described pH doubly in described first time residue, at 20-35 DEG C, stirring 2-4h, filter, collecting second time filtrate and second time residue;
S5, in described second time residue, add 8-12 described pH be doubly the running water of 7.5, stirs 2-4h, filter, collect third time filtrate at 20-35 DEG C;
S6, merge described first time filtrate, second time filtrate and third time filtrate be mixing filtrate, described mixing filtrate is evaporated to the 2-4 of material quantity doubly at 65 DEG C, obtains concentrate;
S7,5-10min is boiled in the heating of described concentrate, add the gluconolactone of concentrated liquid measure 1%-2% while hot, fully dissolve, at 80 DEG C, be incubated 30-60min, then leave standstill more than 5h, protein is fully solidified, then centrifugal or filter, obtain sediment;
S8, described sediment is first washed one time with 75% alcohol, decolouring, then wash one time with 95% alcohol, dehydration, centrifugal or filter and drain, in 55-60 DEG C of vacuum drying, pulverizing, obtains navy bean alpha-amylase inhibitor product.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106310707A (en) * | 2016-08-31 | 2017-01-11 | 青岛佰福得科技有限公司 | Phaseolin extracting method |
| CN110684090A (en) * | 2019-10-09 | 2020-01-14 | 完美（广东）日用品有限公司 | Alpha-amylase inhibitor and preparation process and application thereof |
| CN110973458A (en) * | 2019-11-26 | 2020-04-10 | 同福集团股份有限公司 | Health-care coarse cereal porridge and preparation method thereof |
| CN113318143A (en) * | 2021-05-28 | 2021-08-31 | 德州职业技术学院 | Preparation method of alpha-amylase inhibitor by using beans as raw materials |
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| 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 |
| CN102552376A (en) * | 2010-12-31 | 2012-07-11 | 西安天一生物技术有限公司 | White kidney bean extract extracted by microwave as well as preparation method and application for same |
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| 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 |
| CN102552376A (en) * | 2010-12-31 | 2012-07-11 | 西安天一生物技术有限公司 | White kidney bean extract extracted by microwave as well as preparation method and application for same |
| CN103638089A (en) * | 2013-11-29 | 2014-03-19 | 广东省食品工业研究所 | Method for extracting alpha-amylase inhibitor from white kidney beans by use of microwave-compound enzyme coupling method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106310707A (en) * | 2016-08-31 | 2017-01-11 | 青岛佰福得科技有限公司 | Phaseolin extracting method |
| CN106310707B (en) * | 2016-08-31 | 2018-09-18 | 青岛佰福得科技有限公司 | A kind of extracting method of phaseolin |
| CN110684090A (en) * | 2019-10-09 | 2020-01-14 | 完美（广东）日用品有限公司 | Alpha-amylase inhibitor and preparation process and application thereof |
| CN110973458A (en) * | 2019-11-26 | 2020-04-10 | 同福集团股份有限公司 | Health-care coarse cereal porridge and preparation method thereof |
| CN113318143A (en) * | 2021-05-28 | 2021-08-31 | 德州职业技术学院 | Preparation method of alpha-amylase inhibitor by using beans as raw materials |
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