CN103159839A - Thermostability-protected alpha-amylase inhibitor and application thereof - Google Patents
Thermostability-protected alpha-amylase inhibitor and application thereof Download PDFInfo
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Abstract
The invention relates to a thermostability-protected alpha-amylase inhibitor and application thereof, belonging to the technical field of food additives. The product is prepared by the following steps: (1) preparing a maltitol or mannitol water solution with the mass/volume ratio concentration of 1-10%, regulating alpha-amylase inhibitor dry powder of leguminosae and graminaceous plant seeds into pulp with the water solution, and regulating the pH value to 6.5-7.8 with sodium bicarbonate or hydrochloric acid, wherein the mass ratio of the water solution to the dry powder is 1:(3-8); (2) keeping the pulp at the temperature of 35-55 DEG C under ultrasonic irradiation for 10-40 minutes, wherein the ultrasonic frequency is 40-2000 KHZ, and the ultrasonic intensity is 0.4-6.5 W/cm<2>; and (3) drying the obtained substance to obtain the product. The alpha-amylase inhibitor can be used as an active component of starch food additives. The alpha-amylase inhibitor is pretreated, so that the glycoprotein structure is protected, thereby enhancing the high-temperature stability of the alpha-amylase inhibitor. The alpha-amylase inhibitor can be added into starch before the heating procedure of preparing starch food.
Description
Technical field
The invention belongs to technical field of food additives, be specifically related to product that the Alpha-starch inhibitor from pulse family and grass seed is carried out gained after thermally-stabilised conservation treatment and uses thereof.
Background technology
Since the seventies, studied and found more than the 100 kind of active substance from the inhibition α-amylase of plant and microorganism.The investigator has explored the regularity of distribution of alpha-amylase inhibitor in the different fruit of starch content, the alpha-amylase inhibitor of different plants and the relation between starch content have been compared, result draws alpha-amylase inhibitor content and Starch Contents In Plants has strong dependency, be in the higher fruit of starch content, its contained alpha-amylase inhibitor is also higher.Alpha-amylase inhibitor can separate from wheat, barley, millet, Chinese sorghum, cereal, peanut, soybean, other plant of taro root and some.The inhibitor molecules weight range is 9KD ~ 63KD.The alpha-amylase inhibitor of plant origin can with α-amylase generation specific binding, thereby hinder digestion and the decomposition of carbohydrate in food, effectively reduce body and obtain the heat that produces because of starch food digestion.Due to alpha-amylase inhibitor only in Digestive tract with the Digestive system effect, do not enter blood, do not act on human organ, therefore can be used as normal food nutrition ancillary component and add, be used for prevention and treatment obesity and diabetes.Alpha-amylase inhibitor companion canteen is with inconvenient, and if make the danger that pulvis sucks respiratory tract in addition; This inhibitor itself is a kind of glycoprotein molecule structure, its activeconstituents has thermolability, heating was over 5 minutes in the above temperature at 70 degrees centigrade, glycoprotein structure will be by havoc, thereby cause glycoprotein to lose diastatic noncompetitive inhibit feature, therefore should not be used for adding starch to make starch food products by heating.The Alpha-starch inhibitor of plant origin mostly is before the meal greatly and directly takes at present, but this is taken mode and easily causes inhibitor effect low, because people's digestive juice secretion all exists in whole feed process, and diastatic secretion produces with feed, take in advance cause amylase inhibitor can not follow diastatic secretion suppress the blocking-up, this reduces the inhibitor effect greatly.
" protection of N.F,USP MANNITOL to alpha-amylase inhibitor " literary composition [" Anhui medicine ", 2007 Jul.; II (7)] report, find after deliberation, add people's N.F,USP MANNITOL in a amylase inhibitor water extraction liquid is carried out drying process, in the dry powder that obtains, the active not reduction of the inhibition of amylase inhibitor has raise a lot on the contrary.But find in our repetition test, directly add N.F,USP MANNITOL to exist obvious use problem in the amylase inhibitor aqueous solution.At first N.F,USP MANNITOL can only join in the starch inhibitor aqueous solution after dissolving, in the extracting solution of perhaps handling well, caused like this content of the amylase inhibitor that contains in liquid extremely low, and the N.F,USP MANNITOL amount that adds is very high, the feasibility of its suitability for industrialized production is lower, and cost is higher; Secondly do not adding in hyperacoustic situation, its protection efficient to inhibitor is relatively low; Moreover after the long-time heat of the amylase inhibitor that document method is processed through preparation food the time, the inhibition activity of amylase inhibitor has greater loss.
Summary of the invention
The object of the invention is to provides alpha-amylase inhibitor of a kind of thermally-stabilised protection and uses thereof for overcoming the deficiencies in the prior art.
The alpha-amylase inhibitor of the thermally-stabilised protection of the present invention is the product that makes as follows:
1, the preparation quality volume by volume concentration is 1%～10% maltose alcohol or the aqueous solution of N.F,USP MANNITOL, to be adjusted to slurry from the Alpha-starch inhibitor dry powder of pulse family and grass seed with this aqueous solution, the mass ratio of the said aqueous solution and dry powder is 1:3～8, and transferring pH value with sodium bicarbonate or hydrochloric acid is 6.5-7.8;
2, slurry is being incubated 10-40 minutes under ul-trasonic irradiation under 35-55 degree centigrade, ultrasonic frequency is 40 ~ 2000KHZ, and ultrasonic intensity is 0.4 ~ 6.5 watt/every square centimeter;
3, gains are carried out drying, make the invention product.Drying means preferably sprays dried.
Mass volume ratio concentration is 1%～10% the aqueous solution, and 1 ~ 10g solute is dissolved in 100ml water exactly.
Intensity of acoustic wave refers in the unit time, and sound wave is by the acoustic energy perpendicular to the propagation direction unit surface.
The present invention is by to α--and the starch inhibitor is processed in advance, and its glycoprotein structure is protected, thus reinforcing alpha--the stability under starch inhibitor high temperature.
The alpha-amylase inhibitor of the thermally-stabilised protection of the present invention is used for the activeconstituents as the starch food additive.Said starch food additive can be the product of the present invention of single component, it can be also the composition of product of the present invention and receptible other compositions of food additive, said starch food additive can join starch before the heating process of preparation starch food in, also can join in the starch food of having made, but in preferably joining starch before the heating process of preparation starch food, so just can demonstrate fully the heat-resisting advantage of product of the present invention.
Below that the method that the amylase inhibitor through the thermostability conservation treatment adds in starch food is given an example:
1, boiling class,
A, fermentation class: after fermentation ends, in the dough-kneading process, amylase inhibitor is mixed into dry flour, carries out kneading, usage ratio: 1%～20% of the total consumption of flour (weight that comprises bread dough).
B, non-fermentation class: inhibitor is first transferred with warm water melted, then the water that mixes up is joined in flour, carry out kneading.Usage ratio: 0.5%～20% of the total consumption of flour.
Other program of boiling class is carried out according to other normal process program of food, does not affect making processes.(steaming temperature and steaming time are by normally steaming operating of wheaten food)
2, baking class
A, fermentation class: be added into before fermentation in dry flour, ferment with flour, perhaps after fermentation ends, in the dough-kneading process, amylase inhibitor is mixed into dry flour, carries out kneading, usage quantity: 1%～12% of the total consumption of flour (weight that comprises bread dough).
The non-fermentation class of B: inhibitor is first transferred with warm water melted, then the water that mixes up is joined in flour, carry out kneading.Usage ratio: 0.5%～12% of the total consumption of flour.
Other auxiliary material and making processes are all completed according to the normal process program.
3, other class
Amylase inhibitor that can thermal treatment is good adds to preparation in bulk, and is directly edible in the heat foods such as potato class as rice, hot French fries, baking.
We contrast by animal experiment, have confirmed that product of the present invention has good thermostability, and existing alpha-amylase inhibitor is not had a thermostability.Said amylase inhibitor is the Alpha-starch inhibitor from pulse family and grass seed.
Test one: the Different treatments amylase inhibitor is to rat prevention of obesity effect.
Experimental animal: choose the male SD rat of 120g ± 20g, be divided into five groups, the cycle of carrying out is 90 days feeding.
Blank group: the normal diet of feeding experiment mouse.
The high lipid food group: increase in mass ratio by 5% grease on the basis of normal diet, 5% sucrose, 10% protein is 120 degrees centigrade of oven drying half an hour.
The amylase inhibitor group that is untreated (namely existing amylase inhibitor group): 1% ratio is added untreated amylase inhibitor in mass ratio in high lipid food, 120 degrees centigrade of oven drying half an hour.
The treatment with mannitol group of product of the present invention: 1% ratio is added the product of the present invention that obtains through treatment with mannitol in mass ratio in high lipid food.The preparation method of product of the present invention used: 5g N.F,USP MANNITOL is dissolved in 100ml water after mixing, add 500g amylase inhibitor sterling, transfer to pulpous state, be placed in the hyperacoustic water-bath of 200KHz 45 ℃ of heating 20 minutes, ultrasonic intensity is 1.6 watts/every square centimeter; And in this process with sodium bicarbonate adjust pH to 7.0, carry out the drying of baking oven half an hour after completing under 120 degrees centigrade, standby.
The maltose alcohol treatment group of product of the present invention: 1% ratio is added the product of the present invention that processing obtains through maltose alcohol in mass ratio in high lipid food.The preparation method of product of the present invention used: 5g maltose is dissolved in 100ml water after mixing, add 500g amylase inhibitor sterling, transfer to pulpous state, be placed in the hyperacoustic water-bath of 200KHz 45 ℃ of heating 20 minutes, ultrasonic intensity is 1.6 watts/every square centimeter; And in this process with sodium bicarbonate adjust pH to 7.0, carry out the drying of baking oven half an hour after completing under 120 degrees centigrade, standby.
Male SD obtains following testing data after DIFFERENT FEED was fed 90 days.
A, the indices of male SD rat in the fat model of getting fat is fed process
The body weight index
Annotate: P＜0.01, there were significant differences is denoted as * with the blank group; There were significant differences is denoted as with the high lipid food group
▲
The fat pad weightening finish of testis week
The rat total cholesterol level
Annotate: P＜0.01, there were significant differences is denoted as * with the blank group, and there were significant differences is denoted as with the high lipid food group
▲
According to above test-results, reach a conclusion: the amylase inhibitor that process maltose alcohol or N.F,USP MANNITOL add ultrasonication still has the effect of prevention of obesity after heating, but add undressed amylase inhibitor feed and common high lipid food all not to possess the prevention of obesity effect
Testing two Different treatments amylase inhibitors tests the rat effect of weight reducing
Experimental animal: choose the modeling success male SD rat, be divided at random 4 groups, the cycle of carrying out is that the Different treatments feed of 45 days is fed.
Blank group: the common high lipid food of the modeling of feeding.
The amylase inhibitor group is untreated: the ratio in 1% in high lipid food is added untreated amylase inhibitor, oven drying (120 degrees centigrade of lower half an hour).
The treatment with mannitol group: the ratio in 1% in high lipid food is added the amylase inhibitor that obtains through treatment with mannitol.The treatment with mannitol method: 5g N.F,USP MANNITOL is dissolved in 100ml water after mixing, adds 500g amylase inhibitor sterling, transfers to pulpous state, is placed in the hyperacoustic water-bath of 200KHz 45 ℃ of heating 20 minutes, and ultrasonic intensity is 1.6 watts/every square centimeter; And in this process with sodium bicarbonate adjust pH to 7.0, oven drying after completing (120 degrees centigrade of lower half an hour).
Maltose alcohol treatment group: add through maltose alcohol in 1% ratio in high lipid food and process the amylase inhibitor that obtains.
The maltose alcohol treatment process: the 5g maltose alcohol is dissolved in 100ml water after mixing, adds 500g amylase inhibitor sterling, transfers to pulpous state, is placed in 200KHz ultrasound bath pot 45 ℃ of heating 20 minutes, and ultrasonic intensity is 1.6 watts/every square centimeter; And in this process with dilute hydrochloric acid adjust pH to 7.0, oven drying after completing (120 degrees centigrade of lower half an hour).
Indices in male SD rat fat-reducing test:
The body weight index
Annotate: P＜0.01, there were significant differences is denoted as * with the blank group.
The fat pad weightening finish of testis week
Annotate: P＜0.01, there were significant differences is denoted as * with the blank group.
The rat triglyceride
Annotate: P＜0.01, there were significant differences is denoted as * with the blank group.
The rat cholesterol
Annotate: P＜0.01, there were significant differences is denoted as * with the blank group.
According to above test-results, reach a conclusion: add through maltose alcohol or N.F,USP MANNITOL the amylase inhibitor that ultrasonication obtains and still have fat-reducing effect after heating, but undressed amylase inhibitor is not had a fat-reducing effect
Measured under the heating environment of 160 degrees centigrade be untreated, treatment with mannitol and maltose alcohol process the AIA loss ratio that obtains, and the results are shown in Figure 1.
Also the AIA of non-ultrasonication and ultrasonication has been done simultaneous test.Test conditions is:
Non-ultrasonication group: the maltose alcohol with the pure and mild homogenous quantities of 3g N.F,USP MANNITOL adds water 30ml dissolving respectively, solution is joined in the amylase inhibitor dry powder 100g of Semen Phaseoli Vulgaris extraction, is adjusted to pulpous state and adjusts pH to 7.0 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in respectively the container water-bath is heated to 45 ℃ and be incubated 20 minutes; Taking out spray does.
The ultrasonication group: the maltose alcohol with the pure and mild homogenous quantities of 3g N.F,USP MANNITOL adds water 30ml dissolving respectively, solution is joined in the amylase inhibitor dry powder 100g of Semen Phaseoli Vulgaris extraction, be placed on respectively in the hyperacoustic water-bath of 200KHz 45 ℃ of heating 20 minutes, ultrasonic intensity is 1.6 watts/every square centimeter; And in this process with sodium bicarbonate adjust pH to 7.0, take out spray and do.
Do not process group: directly take the amylase inhibitor dry product of not doing any processing.
Get and respectively organize sample and survey it suppress active after 160 ℃ of heating, the results are shown in Figure 2 and Fig. 3.
Beneficial effect of the present invention: a kind of amylase inhibitor of Heat stability is good is provided, can have joined starch before the heating process of preparation starch food.
Description of drawings
Fig. 1 is the loss of activity comparison diagram that is untreated, adds the ultrasonic wave treatment with mannitol, adds 160 ℃ of heating of amylase inhibitor of ultrasonic wave maltose alcohol processing.
Fig. 2 is the loss of activity comparison diagram that is untreated, does not add the ultrasonic wave treatment with mannitol, adds 160 ℃ of heating of amylase inhibitor of ultrasonic wave treatment with mannitol.
Fig. 3 is untreated, does not add the loss of activity comparison diagram that 160 ℃ of heating of amylase inhibitor of ultrasonic wave maltose alcohol processing were processed, added to the ultrasonic wave maltose alcohol.
Embodiment
Example 1, get 20g N.F,USP MANNITOL and add water 200ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that wheat seed extracts, be adjusted to pulpous state and adjust PH to 7.8 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 35 ℃ and be incubated 40 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 0.4 watt/every square metre; Take out spray and do, 160 ℃ of heating 30 minutes, recording the starch enzyme inhibition activity after cooling is 2120U/g.
Example 2, get 5g N.F,USP MANNITOL and add water 500ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that the kidney bean seed extracts, be adjusted to pulpous state and adjust PH to 6.8 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in container, is heated to 55 ℃ and be incubated 10 minutes in the mode of water-bath in the ultrasound bath pot, ultrasonic intensity is 6.5 watts/every square metre; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 1930U/g.
Example 3, get 2g N.F,USP MANNITOL and add water 200ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that wheat seed extracts, be adjusted to pulpous state and adjust PH to 6.8 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in container, is heated to 25 ℃ and be incubated 5 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 520U/g.
Example 4, get 2g N.F,USP MANNITOL and add water 200ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that wheat seed extracts, be adjusted to pulpous state and adjust PH to 6.5 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, 160 ℃ of heating 30 minutes, recording the starch enzyme inhibition activity after cooling is 920U/g.
Example 5, get 2g N.F,USP MANNITOL and add water 200ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that wheat seed extracts, be adjusted to pulpous state and adjust PH to 6.5 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in container, is heated to 25 ℃ and be incubated 40 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, 160 ℃ of heating 30 minutes, recording the starch enzyme inhibition activity after cooling is 1377U/g.
Example 6, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that wheat seed extracts, be adjusted to pulpous state and adjust PH to 7.8 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 35 ℃ and be incubated 10 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, 160 ℃ of heating 30 minutes, recording the starch enzyme inhibition activity after cooling is 867U/g.
Example 7, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.0 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in container, is heated to 35 ℃ and be incubated 40 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 1520U/g.
Example 8, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.0 with dilute hydrochloric acid; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 10 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square metre; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 989U/g.
Example 9, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.3 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 200KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 740U/g.
Example 10, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.3 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 40KHz ultrasound bath pot, ultrasonic intensity is 0.4 watt/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 667U/g.
Example 11, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.3 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 1000KHz ultrasound bath pot, ultrasonic intensity is 0.4 watt/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 940U/g.
Example 12, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.0 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 2000KHz ultrasound bath pot, ultrasonic intensity is 0.4 watt/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 446U/g.
Example 13, get the 30g maltose alcohol and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g that Semen Phaseoli Vulgaris extracts, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 2000KHz ultrasound bath pot, ultrasonic intensity is 6.5 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 97U/g.
Example 14: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of wheat extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 2000KHz ultrasound bath pot, ultrasonic intensity is 6.5 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 261U/g.
Example 15: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of Semen Phaseoli Vulgaris extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 2000KHz ultrasound bath pot, ultrasonic intensity is 6.5 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 192U/g.
Example 16: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of Semen Phaseoli Vulgaris extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 40KHz ultrasound bath pot, ultrasonic intensity is 6.5 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 692U/g.
Example 17: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of Semen Phaseoli Vulgaris extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 1000KHz ultrasound bath pot, ultrasonic intensity is 6.5 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 865U/g.
Example 18: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of Semen Phaseoli Vulgaris extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 40KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 2191U/g.
Example 19: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of Semen Phaseoli Vulgaris extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 1000KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 1168U/g.
Example 20: get 30g N.F,USP MANNITOL and add water 300ml dissolving, solution is joined in the amylase inhibitor dry powder 1000g of Semen Phaseoli Vulgaris extraction, be adjusted to pulpous state and adjust PH to 7.1 with sodium bicarbonate; The soup compound that mixes up PH is placed in container, is heated to 50 ℃ and be incubated 40 minutes in the mode of water-bath in 2000KHz ultrasound bath pot, ultrasonic intensity is 1.6 watts/every square centimeter; Take out spray and do, recording the starch enzyme inhibition activity after 160 ℃ of heating 30 minutes are cooling is 498U/g.
The amylase that the above provides suppresses activity value and measures by the following method:
The alpha-amylase inhibitor activity determination method
1. the scope of application
Present method is measured the activity from the Alpha-starch inhibitor of pulse family and grass seed.
2. principle
Alpha-amylase inhibitor changes the conformation of α-amylase, and then causes α-amylase to lose diastatic ability by forming the complex body of solubility with α-amylase.Utilize starch solution aobvious blue under the IKI effect, in certain starch concentration scope, the light absorption value at 660nm place and the linear characteristics of the amount of starch, then according to the difference that adds α-amylase starch-splitting amount before and after alpha-amylase inhibitor, can calculate the activity of inhibitor.
3. term and definition
Following term and definition are applicable to this standard.
3.1 α-amylase alpha-amylase
1g alpha-amylase inhibitor powder under 37 ℃, PH=6.0 condition, reduces the amount that α-amylase digestion forms maltose in 1min, be 1 enzyme inhibitors unit of activity, with U/g.
3.2 alpha-amylase inhibitor
Special glycoprotein structure can the glucosides shearing site on the α-amylase molecule be combined and causes that the α-amylase molecular conformation changes, and is alpha-amylase inhibitor thereby make it lose the compound that the ability of being combined with the starch glucosides causes catalytic activity to be lost.
3.3 alpha-amylase inhibitor vigor
1mg alpha-amylase inhibitor powder under 37 ℃, PH=6.0 condition, suppresses the amount that α-amylase consumes starch in 1min, be 1 enzyme inhibitors unit of activity, with U/mg.
4. reagent preparation
4.1 Zulkovsky starch solution (2g/L): take 0.200g(and be accurate to 0.001g) Zulkovsky starch (in over dry) is in beaker, with a small amount of water furnishing soup compound, slowly add while stirring in 70ml boiling water, then the beaker of dress starch is rinsed in the water gradation, washing lotion is poured into wherein, be heated with stirring to fully transparent, the cooling 100ml that is settled to.Solution is now with the current.
4.2 phosphoric acid buffer (pH=6.0): take 45.23g Sodium phosphate dibasic (Na
2HPO
412H
2O) and 8.07g citric acid (C
6H
8O7H
2O), with water dissolution and be settled to 1000ml.Use after proofreading and correct with pH meter.
4.3 former iodine liquid: take 11.0g iodine and 22.0g potassiumiodide, with a small amount of water, iodine is dissolved fully, be settled to 500ml, be stored in brown bottle.
4.4 rare iodine liquid: draw former iodine liquid 2.00ml, add the 20.0g potassiumiodide with water dissolution and be settled to 500ml, be stored in brown bottle.
4.5 hydrochloric acid soln (0.1mol/L): get hydrochloric acid 9ml, add water and make in right amount 100ml, shake up.
4.6 Porcine amylopsin: press GB 8275-2009 and measure active.Be stored in refrigerator.
5. instrument and equipment
5.1 spectrophotometric is taken into account cuvette.
5.2 electronic analytical balance.
5.3 standard ground band plug volumetric flask: 100ml, 500ml.
5.4 thermostat water bath: temperature-controlled precision ± 0.1 ℃
5.5 test tube: 25 mm * 200 mm.
5.6 standard transfer pipet (2 ml, 10 ml), micropipette rifle.
5.7 stopwatch.
5.8 pH meter: scale division value 0.05.
5.9 whizzer and centrifuge tube.
5.10 ultrasonic washing instrument.
Use-case 1, the wheat starch enzyme inhibitors 100g that hot conservation treatment is complete add in the 10kg whole meal flour after being dissolved in water, and after adding yeast fermentation, get bread dough and make bread, and normally smoke the program baking by bread.
Use-case 2, the kidney bean amylase inhibitor 400g dry powder that hot conservation treatment is complete are blended in the 1600g whole meal flour, add yeast powder, after adding water kneading and fermentation, get bread dough and make bread, and normally smoke the program baking by bread.
Use-case 3, the wheat starch enzyme inhibitors 120g dry powder that hot conservation treatment is complete are blended in the 880g whole meal flour, add egg, water, and other batching kneading dough such as essence becomes the biscuit shape shape with finishing, normal baking to being fit to hardness.
Use-case 4, the kidney bean amylase inhibitor 10g dry powder that hot conservation treatment is complete are blended in the 2000g whole meal flour, add egg, water, and other batching kneading dough such as essence becomes the biscuit shape shape with finishing, normal baking to being fit to hardness.
Use-case 5, the kidney bean amylase inhibitor 100g that hot conservation treatment is complete add in the 10kg whole meal flour after being dissolved in water, and after adding yeast fermentation, get bread dough and cook steamed bun, cook by the pattern that normally steams with steam to get final product.
Use-case 6, the wheat starch enzyme inhibitors 200g that hot conservation treatment is complete add in the 800g whole meal flour after being dissolved in water, and after adding yeast fermentation, get bread dough and cook the steamed stuffed bun musculus cutaneus, cook by the pattern that normally steams with steam after faric material to get final product.
Use-case 7, the kidney bean amylase inhibitor 100g dry powder that hot conservation treatment is complete are blended in the 20kg whole meal flour after being dissolved in water; add egg, water; other batching kneading dough such as essence becomes noodles to being fit to hardness with finishing, normally gets final product after drying.
Use-case 8, wheat starch enzyme inhibitors 200g dry powder mixing that hot conservation treatment is complete get final product according to traditional vermicelli manufacturing procedure addition vermicelli in 800g Ipomoea batatas flour.
Use-case 9, the wheat starch enzyme inhibitors 1g that hot conservation treatment is complete add in the 100g fried potato directly edible with the dusting pattern.
Use-case 10, the kidney bean amylase inhibitor 1g that hot conservation treatment is complete add in 100g rice directly edible in the dusting mode.
Claims (5)
1. the alpha-amylase inhibitor of a thermally-stabilised protection is characterized in that for making as follows:
(1), the preparation quality volume by volume concentration is 1%～10% maltose alcohol or the aqueous solution of N.F,USP MANNITOL, to be adjusted to slurry from the Alpha-starch inhibitor dry powder of pulse family and grass seed with this aqueous solution, the mass ratio of the said aqueous solution and dry powder is 1:3～8, and transferring pH value with sodium bicarbonate or hydrochloric acid is 6.5-7.8;
(2), slurry is being incubated 10-40 minutes under ul-trasonic irradiation under 35-55 degree centigrade, ultrasonic frequency is being 40 ~ 2000KHZ, and ultrasonic intensity is 0.4 ~ 6.5 watt/every square centimeter;
(3), gains are carried out the dry product that gets.
2. the alpha-amylase inhibitor of thermally-stabilised protection as claimed in claim 1, the method that it is characterized in that said drying is that spray is done.
3. the purposes of the alpha-amylase inhibitor of thermally-stabilised protection as claimed in claim 1, is characterized in that the activeconstituents as the starch food additive.
4. the purposes of the alpha-amylase inhibitor of thermally-stabilised protection as claimed in claim 3, is characterized in that said starch food additive is single component.
5. the purposes of the alpha-amylase inhibitor of thermally-stabilised protection as claimed in claim 4, is characterized in that said starch food additive joined in starch before the heating process of preparation starch food.
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| CN107531747A (en) * | 2015-04-03 | 2018-01-02 | 罗盖特公司 | The stabilization of protein |
| CN113197241A (en) * | 2021-04-22 | 2021-08-03 | 浙江维京生物科技有限公司 | White kidney bean sugar-control biscuit with high-activity amylase inhibition capability and preparation method thereof |
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| JP2001299242A (en) * | 2000-04-28 | 2001-10-30 | Kanebo Ltd | Food containing gelling agent and method for producing the same |
| CN1772216A (en) * | 2005-10-21 | 2006-05-17 | 南京工业大学 | Solid mixture containing alpha-amylase inhibitor, preparation process and application thereof in pharmacy |
| EP1967200A1 (en) * | 2007-03-07 | 2008-09-10 | Indena S.P.A. | Formulations of alpha-amylase inhibitors of P. vulgaris with alpha-glucosidade inhibitores of S. oblonga or S. reticulata useful in the treatment of diabetes and obesity |
| CN101514227A (en) * | 2009-03-20 | 2009-08-26 | 云南天保桦生物资源开发有限公司 | Extraction process for amylase inhibiting protein |
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| JP2001299242A (en) * | 2000-04-28 | 2001-10-30 | Kanebo Ltd | Food containing gelling agent and method for producing the same |
| CN1772216A (en) * | 2005-10-21 | 2006-05-17 | 南京工业大学 | Solid mixture containing alpha-amylase inhibitor, preparation process and application thereof in pharmacy |
| EP1967200A1 (en) * | 2007-03-07 | 2008-09-10 | Indena S.P.A. | Formulations of alpha-amylase inhibitors of P. vulgaris with alpha-glucosidade inhibitores of S. oblonga or S. reticulata useful in the treatment of diabetes and obesity |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107531747A (en) * | 2015-04-03 | 2018-01-02 | 罗盖特公司 | The stabilization of protein |
| CN113197241A (en) * | 2021-04-22 | 2021-08-03 | 浙江维京生物科技有限公司 | White kidney bean sugar-control biscuit with high-activity amylase inhibition capability and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103159839B (en) | 2014-07-30 |
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