CN116042746A - Method for preparing resistant dextrin by debranching and crystallization - Google Patents
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Abstract
The invention relates to the technical field of starch deep processing, in particular to a method for preparing resistant dextrin by debranching and crystallization, which comprises the following steps: gelatinizing starch at high temperature; then pullulanase is used for enzymolysis to remove amylopectin, namely, partial alpha-1, 4 glycosidic bonds in starch are broken, new alpha-1, 2 and alpha-1, 3 glycosidic bonds connected with each other are regenerated, and partial reduction ends are subjected to intramolecular dehydration to form dextran shrink, beta-1, 6 glycosidic bonds and the like, so that the resistant dextrin has digestion resistance; and then adopting different debranching and crystallization methods to prepare the resistant dextrin; finally, the porcine pancreatic alpha-amylase is used for enzyme hydrolysis and purification of the resistant dextrin. The whole process has mild condition, no chemical reagent addition and wastewater generation, no pollution in the production process and environmental friendliness, and the prepared resistant dextrin has obviously increased content compared with the resistant starch of the original starch.
Description
Technical Field
The invention belongs to the technical field of starch deep processing, and particularly relates to a method for preparing resistant dextrin by debranching and crystallization.
Background
The industry of nutritious health foods has becomeThe dietary fiber has various effects of reducing energy value, improving taste, enhancing texture and the like, can be used as a novel food additive auxiliary material or nutrient to be directly added into food to prepare nutritional health food for general or special people, and becomes a hotspot of current food industry development. The resistant dextrin is used as a low molecular water-soluble dietary fiber, can be added into food as a food raw material to play the role of the dietary fiber, does not influence the taste, and has good processing performance. In addition, the resistant dextrin also has physiological activities of reducing blood sugar, balancing the environment in the intestinal tract, promoting the generation of short chain fatty acid in the intestinal tract and the like
M,
K.Efficiency of Resistant Starch and Dextrins as Prebiotics:AReview of the Existing Evidence and Clinical Trials[J].Nutrients,2021,13(11):3808；Kilua A,Pelpolage S,Goto A,et al.Deciphering the colonic fermentation characteristics of agavin and digestion-resistant maltodextrin in a simulated batch fermentation system[J].International Journal of Biological Macromolecules,2021,189:151-159；Hobden M R,Commane D M,Guérin-Deremaux L,et al.Correction to:Impact of dietary supplementation with resistant dextrin/>
on satiety,glycaemia,and related endpoints,in healthy adults[J]European Journal of Nutrition,2021,60 (8): 4645.) is considered as a potential prebiotic and has good application prospect.
The existing preparation methods of the resistant dextrin mainly comprise an acid heating method, a microwave method, an enzyme method and the like, and the earliest and most commonly used preparation method of the resistant dextrin is the acid heating method. It is mainly prepared by preparing pyrodextrin, soaking starch with hydrochloric acid/other acid at high temperature, and then performing a series of operations such as liquefying, enzymatic hydrolysis, decoloring, filtering, concentrating, spray drying and the like to obtain a resistant dextrin product (Kamonrat Trithavisup, kuakarun Krusong, kanitha Tananuwong, in-depth study of the changes In properties and molecular structure of cassava starch during resistant dextrin preparation, food Chemistry,2019, 297:124996). However, most of the acid heating methods have the problems of low purity, dark color, time and energy consumption, environmental pollution and the like. The physical methods such as radiation, microwaves and the like can greatly shorten the preparation time, and can also improve the problem of non-uniformity in the heating process of starch, but the yield and purity are still to be improved. The enzyme method makes up the problems of deep color, environmental pollution and the like of the product, and still needs to further improve the yield and purity.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention aims to provide a method for preparing resistant dextrin by debranching and crystallization, which is used for solving the problems of high process energy consumption, environment friendliness, uneven reaction, low product purity and low activity in the current preparation methods of most resistant dextrins.
The aim of the invention is realized by adopting the following technical scheme:
a method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
step 1, weighing starch, adding the starch into a sodium acetate buffer solution in a dispersing way, and heating and stirring the starch in boiling water to realize gelatinization;
step 2, cooling the gelatinized starch milk in the step 1, and adding pullulanase for enzymolysis;
step 3, debranching and crystallizing the starch milk containing pullulanase in the step 2 to obtain a crude product of resistant dextrin;
step 4, carrying out boiling water bath digestion and cooling on the crude product of the resistant dextrin obtained in the step 3, and then adjusting the pH value to 6.9 to obtain a resistant dextrin sample; the method comprises the steps of carrying out a first treatment on the surface of the
Step 5, adding porcine pancreatic alpha-amylase into the resistant dextrin sample obtained in the step 4 for enzymatic hydrolysis, and placing the mixture at 37 ℃ for stirring and heat preservation;
and 6, centrifuging, freeze-drying and crushing the resistant dextrin subjected to enzymolysis in the step 5 to obtain the resistant dextrin.
Preferably, in the step 1, the starch is common corn starch or waxy corn starch; the common corn starch is starch prepared by the steps of crushing, sieving, precipitating, drying, grinding and the like after corn is soaked by 0.3 percent sulfurous acid, and the content of amylopectin is 20 to 35 percent; waxy corn starch is a polysaccharide substance obtained by wet milling and extracting waxy corn, and the content of amylopectin is more than 95%.
Preferably, in said step 1, the concentration of starch in the buffer solution is 0.2g/ml.
Preferably, in the step 1, the sodium acetate buffer solution is a buffer solution containing 0.02% sodium azide, and the gelatinization time is 30min.
Preferably, in the step 2, the cooling is performed at room temperature and is cooled to a temperature of 50 ℃; the amount of pullulanase added was 1.25mg.
Preferably, in the step 3, the conditions of the debranching and crystallization method are any one of the following: (1) Heating at 50deg.C for 24 hr, and standing in refrigerator at 4deg.C for 7 days; (2) continuously magnetically stirring for 7 days in a water bath kettle at 50 ℃; (3) The mixture was heated and stirred continuously in a homogeneous reactor at 50℃for 7 days.
Preferably, in the step 4, the boiling water bath is steamed for 10min and cooled to 60 ℃.
Preferably, in the step 5, the stirring is magnetic stirring, and the heat preservation time is 3-12h.
The beneficial effects of the invention are as follows:
the invention provides an environment-friendly preparation method of resistant dextrin, which prepares the resistant dextrin by adopting gelatinization, enzymolysis, debranching, crystallization and enzymatic hydrolysis and purification thereof, and researches the influence on the digestibility of the resistant dextrin by adjusting the conditions of raw materials and different debranching and crystallization. Compared with other preparation methods, the method has mild conditions in the whole process, and no chemical reagent is added and wastewater is generated; therefore, the production process is pollution-free and environment-friendly, and the content of the prepared resistant dextrin is obviously increased compared with that of the original starch.
Detailed Description
The technical features, objects and advantages of the present invention will be more clearly understood from the following detailed description of the technical aspects of the present invention, but should not be construed as limiting the scope of the invention.
The starting materials, reagents or apparatus used in the following examples are all available from conventional commercial sources or may be obtained by methods known in the art unless otherwise specified.
The invention will be further described with reference to the following examples.
Example 1
A method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
(1) Respectively weighing a proper amount of waxy corn starch, adding a sodium acetate buffer solution containing 0.02% sodium azide to a concentration of 0.2g/ml, heating and stirring in boiling water, gelatinizing for 30min, and cooling to 50 ℃;
(2) Adding commercial pullulanase into the starch solution obtained in the step (1), heating at 50 ℃ for 24 hours, and then placing in a refrigerator at 4 ℃ for 7 days;
(3) Taking out the crude resistant dextrin product in the step (2) from a refrigerator, steaming in a boiling water bath for 10min, cooling to 60 ℃, and then adjusting the pH to 6.9;
(4) Adding porcine pancreatic alpha-amylase into the sample obtained in the step (3) for enzymatic hydrolysis, and placing the sample at 37 ℃ for magnetic stirring and heat preservation for 3-12h;
(5) Centrifuging the resistant dextrin subjected to enzymolysis in the step (4), freeze-drying the precipitate, crushing, and sealing and storing in a dryer.
Example 2:
a method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
(1) Weighing a proper amount of common corn starch respectively, adding a sodium acetate buffer solution containing 0.02% sodium azide to a concentration of 0.2g/ml, heating and stirring in boiling water, gelatinizing for 30min, and cooling to 50 ℃;
(2) Adding commercial pullulanase into the starch solution obtained in the step (1), heating at 50 ℃ for 24 hours, and then placing in a refrigerator at 4 ℃ for 7 days;
(3) Taking out the crude resistant dextrin product in the step (2) from a refrigerator, steaming in a boiling water bath for 10min, cooling to 60 ℃, and then adjusting the pH to 6.9;
(4) Adding porcine pancreatic alpha-amylase into the sample obtained in the step (3) for enzymatic hydrolysis, and placing the sample at 37 ℃ for magnetic stirring and heat preservation for 3-12h;
(5) Centrifuging the resistant dextrin subjected to enzymolysis in the step (4), freeze-drying the precipitate, crushing, and sealing and storing in a dryer.
Example 3:
a method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
(1) Respectively weighing a proper amount of waxy corn starch, adding a sodium acetate buffer solution containing 0.02% sodium azide to a concentration of 0.2g/ml, heating and stirring in boiling water, gelatinizing for 30min, and cooling to 50 ℃;
(2) Adding commercial pullulanase into the starch solution obtained in the step (1), and continuously magnetically stirring for 7 days in a water bath kettle at 50 ℃ to completely debranch and crystallize the starch;
(3) Taking out the crude resistant dextrin product in the step (2) from the water bath, steaming in a boiling water bath for 10min, cooling to 60 ℃, and then adjusting the pH to 6.9;
(4) Adding porcine pancreatic alpha-amylase into the sample obtained in the step (3) for enzymatic hydrolysis, and placing the sample at 37 ℃ for magnetic stirring and heat preservation for 3-12h;
(5) Centrifuging the resistant dextrin subjected to enzymolysis in the step (4), freeze-drying the precipitate, crushing, and sealing and storing in a dryer.
Example 4:
a method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
(1) Weighing a proper amount of common corn starch respectively, adding a sodium acetate buffer solution containing 0.02% sodium azide to a concentration of 0.2g/ml, heating and stirring in boiling water, gelatinizing for 30min, and cooling to 50 ℃;
(2) Adding commercial pullulanase into the starch solution obtained in the step (1), and continuously magnetically stirring for 7 days in a water bath kettle at 50 ℃ to completely debranch and crystallize the starch;
(3) Taking out the crude resistant dextrin product in the step (2) from the water bath, steaming in a boiling water bath for 10min, cooling to 60 ℃, and then adjusting the pH to 6.9;
(4) Adding porcine pancreatic alpha-amylase into the sample obtained in the step (3) for enzymatic hydrolysis, and placing the sample at 37 ℃ for magnetic stirring and heat preservation for 3-12h;
(5) Centrifuging the resistant dextrin subjected to enzymolysis in the step (4), freeze-drying the precipitate, crushing, and sealing and storing in a dryer.
Example 5:
a method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
(1) Respectively weighing a proper amount of waxy corn starch, adding a sodium acetate buffer solution containing 0.02% sodium azide to a concentration of 0.2g/ml, heating and stirring in boiling water, gelatinizing for 30min, and cooling to 50 ℃;
(2) Adding commercial pullulanase into the starch solution obtained in the step (1) to stir uniformly, transferring a sample into a reaction kettle, and placing the reaction kettle in a homogeneous reactor to heat and stir continuously for 7 days at 50 ℃;
(3) Taking out the crude resistant dextrin product in the step (2) from the reaction kettle, steaming in a boiling water bath for 10min, cooling to 60 ℃, and then adjusting the pH to 6.9;
(4) Adding porcine pancreatic alpha-amylase into the sample obtained in the step (3) for enzymatic hydrolysis, and placing the sample at 37 ℃ for magnetic stirring and heat preservation for 3-12h;
(5) Centrifuging the resistant dextrin subjected to enzymolysis in the step (4), freeze-drying the precipitate, crushing, and sealing and storing in a dryer.
Example 6:
a method for preparing resistant dextrins using debranching and crystallization comprising the steps of:
(1) Weighing a proper amount of common corn starch respectively, adding a sodium acetate buffer solution containing 0.02% sodium azide to a concentration of 0.2g/ml, heating and stirring in boiling water, gelatinizing for 30min, and cooling to 50 ℃;
(2) Adding commercial pullulanase into the gelatinized starch obtained in the step (1) until the starch solution is uniformly stirred, transferring a sample into a reaction kettle, and placing the reaction kettle in a homogeneous reactor, and continuously heating and stirring for 7 days at 50 ℃;
(3) Taking out the crude resistant dextrin product in the step (2) from the reaction kettle, steaming in a boiling water bath for 10min, cooling to 60 ℃, and then adjusting the pH to 6.9;
(4) Adding porcine pancreatic alpha-amylase into the sample obtained in the step (3) for enzymatic hydrolysis, and placing the sample at 37 ℃ for magnetic stirring and heat preservation for 3-12h;
(5) Centrifuging the resistant dextrin subjected to enzymolysis in the step (4), freeze-drying the precipitate, crushing, and sealing and storing in a dryer.
Determination of in vitro digestion Performance of resistant dextrins
The content of fast digestible starch (RDS), slow Digestible Starch (SDS) and Resistant Starch (RS) of the 6 resistant dextrins and their raw starches prepared in examples 1 to 6 above was determined by the Englyst method, and the specific determination method was: accurately weighing 0.6g of the 6 resistant dextrins prepared in the examples 1-6 and two starch samples in a 50mL centrifuge tube, adding 20mL sodium acetate buffer solution, fully vortex, uniformly mixing, boiling water bath for 30min, and continuously oscillating and uniformly mixing in the water bath process. Then cooling and placing in a water bath kettle at 37 ℃ for 30min, and magnetically stirring; adding 5mL of mixed enzymatic hydrolysate of porcine pancreatin and amyloglucosidase into a centrifuge tube respectively, shaking and mixing uniformly, taking out 0.25mL of enzymatic hydrolysate respectively at intervals of 20min and 120min, putting the enzymatic hydrolysate into 10mL of 66% ethanol for enzyme deactivation, and then vortex and mixing uniformly. Centrifuging the enzymolysis solution at 4000r/min for 5min, and measuring the glucose content by adopting a GOPOD method.
The glucose content in the corresponding sample was calculated by the following formula:
at = absorbance of the test solution; vt = total volume of test solution; c=concentration of standard glucose (mg/mL); as=absorbance of standard glucose; wt = weight of sample; d=dilution factor 40.
The content of fast digestible starch (Rapidly digestible starch, RDS), slow digestible starch (Slowly digestible starch, SDS) and Resistant Starch (RS) is calculated according to the following formula:
RDS％＝(G20–FG)×0.9
SDS％＝(G120–G20)×0.9
RS％＝[TS–(RDS+SDS)]＝[TS–(G120×0.9)]
g20 Glucose content (mg) of enzymatic hydrolysate for 20min of starch hydrolysis; FG = initial glucose content; g120 Glucose content (mg) of enzymatic hydrolysate for 120min of starch hydrolysis; ts=total starch content of sample (mg).
The in vitro digestibility test was performed on the resistant dextrin samples and the raw starch obtained in examples 1 to 6, and the results are shown in Table 1 below.
Table 1 resistant dextrins and in vitro digestion characteristics of their raw starches according to different methods of preparation
| Sample of | RDS/％ | SDS/％ | RS/％ |
| Waxy corn starch | 93.56±0.00 | 1.34±0.00 | 5.11±0.00 |
| Example 1 | 91.21±1.30 | 2.06±1.63 | 6.74±2.93 |
| Example 3 | 84.90±3.02 | 8.04±3.23 | 6.14±0.24 |
| Example 5 | 75.43±1.84 | 11.64±4.24 | 13.02±4.66 |
| Common corn starch | 89.41±4.31 | 1.39±2.21 | 10.14±2.28 |
| Example 2 | 71.38±3.37 | 7.87±4.41 | 20.75±3.19 |
| Example 4 | 65.16±0.00 | 12.51±0.00 | 22.33±0.00 |
| Example 6 | 60.64±4.65 | 15.43±2.52 | 24.23±2.99 |
As can be seen from Table 1, compared with the original starch, the RS and SDS contents of the resistant dextrin products prepared in examples 1 to 6 are significantly improved, and the RS content obtained by debranching and crystallization in a homogeneous reactor can reach up to 27%; compared with waxy corn starch, the common corn starch has higher RS and SDS content, and the resistant dextrin prepared from the common corn starch has obviously higher SDS and RS content than the waxy corn starch. In combination, the digestion resistance of the resistant dextrins prepared from example 6 is best.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made to the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention.
Claims (8)
1. A process for the preparation of resistant dextrins by debranching and crystallization, characterized in that it comprises the following steps:
step 1, weighing starch, adding the starch into a sodium acetate buffer solution in a dispersing manner, and placing the starch into boiling water for heating and stirring to realize gelatinization;
step 2, cooling the gelatinized starch milk in the step 1, and adding pullulanase for enzymolysis;
step 3, debranching and crystallizing the starch milk containing pullulanase in the step 2 to obtain a crude product of resistant dextrin;
step 4, carrying out boiling water bath digestion and cooling on the crude product of the resistant dextrin obtained in the step 3, and then adjusting the pH value to 6.9 to obtain a resistant dextrin sample;
step 5, adding porcine pancreatic alpha-amylase into the resistant dextrin sample obtained in the step 4 for enzymatic hydrolysis, and placing the mixture at 37 ℃ for stirring and heat preservation;
and 6, centrifuging, freeze-drying and crushing the resistant dextrin subjected to enzymolysis in the step 5 to obtain the resistant dextrin.
2. The method for preparing resistant dextrin by debranching and crystallization according to claim 1, wherein the starch in the step 1 is common corn starch or waxy corn starch; the common corn starch is starch prepared by crushing, sieving, precipitating, drying and grinding after corn is soaked by 0.3% sulfurous acid, and the content of amylopectin is 20% -35%; waxy corn starch is a polysaccharide substance obtained by wet milling and extracting waxy corn, and the content of amylopectin is more than 95%.
3. A method for preparing resistant dextrins by debranching and crystallization according to claim 1, characterized in that the concentration of starch in the buffer solution is 0.2g/ml.
4. The method for preparing resistant dextrin by debranching and crystallization according to claim 1, characterized in that the sodium acetate buffer solution in the step 1 is a buffer solution containing 0.02% sodium azide; the gelatinization time was 30min.
5. The method for preparing resistant dextrins by debranching and crystallization according to claim 1, characterized in that the cooling in step 2 is carried out at room temperature and to a temperature of 50 ℃; the amount of pullulanase added was 1.25mg.
6. The method for preparing resistant dextrin by debranching and crystallization according to claim 1, characterized in that the method conditions of debranching and crystallization in the step 3 are any one of the following: (1) Heating at 50deg.C for 24 hr, and standing in refrigerator at 4deg.C for 7 days; (2) continuously magnetically stirring for 7 days in a water bath kettle at 50 ℃; (3) The mixture was heated and stirred continuously in a homogeneous reactor at 50℃for 7 days.
7. The method for preparing resistant dextrins by debranching and crystallization according to claim 1, characterized in that the boiling water bath cooking time in step 4 is 10min, cooling to 60 ℃.
8. The method for preparing resistant dextrin by debranching and crystallization according to claim 1, wherein the stirring in the step 5 is magnetic stirring, and the time for heat preservation is 3-12 hours.
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