Preparation Method of Amylodextrin

Abstract

The disclosure relates to a preparation method of a amylodextrin and belongs to the technical field of starch chemical modification. According to the method, de-clustering and complexation effects of ultrasonic waves are used to achieve de-clustering of a starch chain and complexation of an amorphous region and an emulsifier, and then -amylase and pullulanase are used to achieve complex enzymolysis. Because the amorphous region and the emulsifier form a complex which is resistant to enzymolysis, the amorphous region is prevented from being destroyed. Finally, dextrins of different molecular weights are separated by a membrane separation method, so as to obtain a amylodextrin product with low polydispersity coefficient and narrow molecular weight distribution, and the starch comprehensive utilization efficiency is increased to 70% or above.

Claims

1. A method of preparing amylodextrin, comprising directly de-clustering starch with ultrasonic waves, adding an emulsifier for ultrasonic complexation, enzymolysis and membrane separation to obtain the amylodextrin.

2. The method according to claim 1, comprising: (1) ultrasonic de-clustering: adding water into waxy corn starch for blending and ultrasonic treatment; (2) ultrasonic complexation: adding an emulsifier into a raw material obtained in step (1) for ultrasonic treatment; (3) enzymolysis: adding a product obtained in step (2) into a buffer for blending, and adding an enzyme for enzymolysis; (4) ethanol extraction: adding an enzymolysis product obtained in step (3) into absolute ethanol to dissolve a complex and the residual emulsifier; (5) membrane separation: performing membrane separation on a product obtained in step (4) to obtain the amylodextrin; and (6) drying: spray drying a product obtained in step (5) to obtain a finished product.

3. The method according to claim 2, wherein in the step (1), water is added into the starch to prepare 5%-10% starch slurry, and the ultrasonic treatment is performed at an ultrasonic treatment power of 500-600 W, and an action time of 5-15 minutes.

4. The method according to claim 2, wherein the emulsifier added in the step (2) is glyceryl monostearate or palmitic acid, and an addition amount of the emulsifier is 3%-5% mass of the starch.

5. The method according to claim 2, wherein in the step (2), the ultrasonic treatment is performed at an ultrasonic treatment power of 500-600 W, a temperature of 40-50 C., and an action time of 20-30 minutes.

6. The method according to claim 2, wherein in the step (3), -amylase and pullulanase are adopted for enzymolysis, an addition amount of pullulanase is 30-40 ASPU/g dry starch, and an addition amount of -amylase is 20-30 ASPU/g dry starch.

7. The method according to claim 2, wherein in the step (5), ultrafiltration membranes used for membrane separation are hollow fiber membranes with molecular weight cutoffs of 5000 Da and 2000 Da for membrane classification.

8. The method according to claim 2, wherein in step (6), an inlet air temperature during spray drying is 140-160 C.

9. The method according to claim 2, comprising: (1) ultrasonic de-clustering: preparing 5%-10% waxy corn starch slurry and stirring evenly for ultrasonic treatment, wherein an ultrasonic working frequency is 25 kHz, a treatment power is 600 W, a temperature is 25 C., an action time is 3 seconds at an interval of 5 seconds, and a total treatment time is 10 minutes; (2) ultrasonic complexation: adding glyceryl monostearate which is 5% mass of the waxy corn starch into the starch slurry after ultrasonic treatment and stirring evenly, wherein the ultrasonic working frequency is 20 kHz, the treatment power is 500 W, the temperature is 40 C., the action time is 3 seconds at an interval of 5 seconds, and the total ultrasonic treatment time is 20 minutes; (3) complex enzymolysis: adjusting a pH value of a starch-glyceryl monostearate complex solution to 6.0 with a phosphate buffer; heating the solution to 50 C. in a water bath; adding medium-temperature -amylase and pullulanase, wherein an addition amount of pullulanase is 30 ASPU/g dry starch, and an addition amount of -amylase is 30 ASPU/g dry starch; hydrolyzing a mixture in a water bath at 50 C. for 3-4 hours; putting the mixture into a boiling water bath for 5 minutes to inactivate medium-temperature -amylase and pullulanase so as to terminate a reaction; and centrifuging to obtain a supernatant; (4) ethanol extraction: adding an enzymolysis product into absolute ethanol to dissolve glyceryl monostearate; (5) membrane separation: performing membrane separation by using hollow fiber membranes with molecular weight cutoffs of 5000 Da and 2000 Da under conditions that a membrane effective region is 0.4 m.sup.2, an operation temperature is 40 C. and an operation pressure is 15 kPa; and (6) drying: spray drying a component obtained after membrane separation to obtain a finished amylodextrin product.

10. The method according to claim 2, comprising: (1) ultrasonic de-clustering: preparing 5%-10% waxy corn starch slurry and stirring evenly for ultrasonic treatment, wherein an ultrasonic working frequency is 25 kHz, a treatment power is 600 W, a temperature is 25 C., an action time is 3 seconds at an interval of 5 seconds, and a total treatment time is 10 minutes; (2) ultrasonic complexation: adding palmitic acid which is 3% mass of the waxy corn starch into the starch slurry after ultrasonic treatment and stirring evenly, wherein the ultrasonic working frequency is 20 kHz, the treatment power is 500 W, the temperature is 40 C., the action time is 3 seconds at an interval of 5 seconds, and the total ultrasonic treatment time is 20 minutes; (3) complex enzymolysis: adjusting a pH value of a starch-palmitic acid complex solution to 6.0 with a phosphate buffer; heating the solution to 50 C. in a water bath; adding medium-temperature -amylase and pullulanase, wherein an addition amount of pullulanase is 40 ASPU/g dry starch, and an addition amount of -amylase is 20 ASPU/g dry starch; hydrolyzing a mixture in a water bath at 50 C. for 3-4 hours; putting the mixture into a boiling water bath for 5 minutes to inactivate medium-temperature -amylase and pullulanase so as to terminate a reaction; and centrifuging to obtain a supernatant; (4) ethanol extraction: adding an enzymolysis product into absolute ethanol to dissolve a complex and residual palmitic acid; (5) membrane separation: performing membrane separation by using hollow fiber membranes with molecular weight cutoffs of 5000 Da and 2000 Da under conditions that a membrane effective region is 0.4 m.sup.2, an operation temperature is 40 C. and an operation pressure is 15 kPa; and (6) drying: spray drying a component obtained after membrane separation to obtain a finished amylodextrin product.

11. An amylodextrin prepared by the method according to claim 1.

12. The amylodextrin according to claim 11, wherein a content of DP10-30 in the amylodextrin is higher than 70%.

Description

DETAILED DESCRIPTION

[0028] The preferred embodiments of the disclosure will be described below. It should be understood that the embodiments are for better explaining the disclosure and are not intended to limit the disclosure.

[0029] 1. The Starch Utilization Rate

[0030] Namely, the yield of amylodextrin is expressed by the ratio of the total mass of all dextrin components after membrane separation to the mass of waxy corn starch used. A calculation formula is as follows:

[0031] Starch utilization rate

[00001]$\left(\%\right)=\frac{\begin{array}{c}\mathrm{Total}.Math..Math.\mathrm{mass}.Math..Math.\mathrm{of}.Math..Math.\mathrm{linear}-\\ \mathrm{chain}.Math..Math.\mathrm{dextrin}.Math..Math.\mathrm{after}.Math..Math.\mathrm{membrane}.Math..Math.\mathrm{separation}.Math..Math.\left(\mathrm{mg}\right)\end{array}}{\mathrm{Mass}.Math..Math.\mathrm{of}.Math..Math.\mathrm{waxy}.Math..Math.\mathrm{corn}.Math..Math.\mathrm{starch}.Math..Math.\left(\mathrm{mg}\right)}*1.Math.0.Math.0.Math.\%$

[0032] 2. Measurement of the Waxy Corn Starch-Emulsifier Complexation Rate

[0033] The complexation rate is expressed by the utilization ratio of a complex to an emulsifier. The content of the emulsifier in the complex is expressed by the amount of the emulsifier washed off in step (4) after the residual emulsifier is washed off in step (2), and the complexation rate is calculated according to the following formula:

[0034] Waxy Corn Starch-Emulsifier Complexation Rate

[00002]$\left(\%\right)=\frac{\mathrm{Emulsifier}.Math..Math.\mathrm{content}.Math..Math.\mathrm{of}.Math..Math.\mathrm{complex}.Math..Math.\left(\mathrm{mg}\right)}{\mathrm{Addition}.Math..Math.\mathrm{amount}.Math..Math.\mathrm{of}.Math..Math.\mathrm{emulsifier}.Math..Math.\left(\mathrm{mg}\right)}*1.Math.0.Math.0.Math.\%$

[0035] 3. Chain Length Distribution of Amylodextrin

[0036] The chain length distribution of amylodextrin is detected by high performance anion exchange chromatography (HPAEC-PAD) equipped with a pulse current detector. The HPAEC-PAD chromatography system is equipped with an ED40 pulsed amperometric detector. The chromatography column model is Dionex CarboPAC PA200 (250*4 mM I.D.). The eluent A is a 100 mmol/L NaOH solution, and the eluent B is a 100 mmol/L NaOH solution containing 600 mmol/L sodium acetate. Linear gradient elution is adopted: a 20% eluent B is used at 0 minute, and a 100% eluent B is used at 60 minutes. The flow rate is 1 ml/min, and the injection volume is 25 l.

[0037] -amylase and pullulanase are purchased from Sigma.

Example 1

[0038] (1) Ultrasonic de-clustering: 5%-10% waxy corn starch slurry is prepared and stirred evenly for ultrasonic treatment, the ultrasonic working frequency is 25 kHz, the treatment power is 600 W, the temperature is 25 C., the action time is 3 seconds at an interval of 5 seconds, and the total treatment time is 10 minutes.

[0039] (2) Ultrasonic complexation: glyceryl monostearate which is 5% mass of the waxy corn starch is added into the starch slurry after ultrasonic treatment and stirred evenly, the ultrasonic working frequency is 20 kHz, the treatment power is 500 W, the temperature is 40 C., the action time is 3 seconds at an interval of 5 seconds, and the total ultrasonic treatment time is 20 minutes.

[0040] (3) Complex enzymolysis: the pH of a starch-glyceryl monostearate complex solution is adjusted to 6.0 with a phosphate buffer, the solution is heated to 50 C. in a water bath, medium-temperature -amylase and pullulanase are added, the addition amount of pullulanase is 30 ASPU/g dry starch, the addition amount of -amylase is 30 ASPU/g dry starch, the mixture is hydrolyzed in a water bath at 50 C. for 3-4 hours, the mixture is put into a boiling water bath for 5 minutes to inactivate medium-temperature -amylase and pullulanase so as to terminate a reaction, and centrifugation is carried out to obtain a supernatant.

[0041] (4) Ethanol extraction: an enzymolysis product is added into absolute ethanol to dissolve glyceryl monostearate.

[0042] (5) Membrane separation: hollow fiber membranes with molecular weight cutoffs of 5000 Da and 2000 Da are used for membrane separation under the conditions that the membrane effective region is 0.4 m.sup.2, the operation temperature is 40 C. and the operation pressure is 15 kPa.

[0043] (6) Drying: a component obtained after membrane separation is spray dried to obtain a finished amylodextrin product.

Example 2

[0044] (1) Ultrasonic de-clustering: 5%-10% waxy corn starch slurry is prepared and stirred evenly for ultrasonic treatment, the ultrasonic working frequency is 25 kHz, the treatment power is 600 W, the temperature is 25 C., the action time is 3 seconds at an interval of 5 seconds, and the total treatment time is 10 minutes.

[0045] (2) Ultrasonic complexation: palmitic acid which is 3% mass of the waxy corn starch is added into the starch slurry after ultrasonic treatment and stirred evenly, the ultrasonic working frequency is 20 kHz, the treatment power is 500 W, the temperature is 40 C., the action time is 3 seconds at an interval of 5 seconds, and the total ultrasonic treatment time is 20 minutes.

[0046] (3) Complex enzymolysis: the pH of a starch-palmitic acid complex solution is adjusted to 6.0 with a phosphate buffer, the solution is heated to 50 C. in a water bath, medium-temperature -amylase and pullulanase are added, the addition amount of pullulanase is 40 ASPU/g dry starch, the addition amount of -amylase is 20 ASPU/g dry starch, the mixture is hydrolyzed in a water bath at 50 C. for 3-4 hours, the mixture is put into a boiling water bath for 5 minutes to inactivate medium-temperature -amylase and pullulanase so as to terminate a reaction, and centrifugation is carried out to obtain a supernatant.

[0047] (4) Ethanol extraction: an enzymolysis product is added into absolute ethanol to dissolve palmitic acid.

[0048] (5) Membrane separation: hollow fiber membranes with molecular weight cutoffs of 5000 Da and 2000 Da are used for membrane separation under the conditions that the membrane effective region is 0.4 m.sup.2, the operation temperature is 40 C. and the operation pressure is 15 kPa.

[0049] (6) Drying: a component obtained after membrane separation is spray dried to obtain a finished amylodextrin product.

Comparative Example 1

[0050] Ultrasonic de-clustering (1) and ultrasonic complexation (2) in Example 1 are omitted, and other conditions or parameters are the same as those in Example 1. This example is equivalent to a blank control. The complexation rate and the starch utilization rate are extremely low, indicating that the waxy corn starch and the emulsifier can hardly form a complex without any external force, and subsequent enzymolysis is not facilitated.

Comparative Example 2

[0051] Ultrasonic de-clustering (1) in Example 1 is omitted, and other conditions or parameters are the same as those in Example 1. Compared with Example 1, the complexation rate and the starch utilization rate are reduced to 32% and 37% respectively, because the starch chain is not fully stretched but is still in an aggregated state, which hinders complexation of the amorphous region and the emulsifier. Therefore, ultrasonic treatment in step (1) has the effect of de-clustering the starch chain.

Comparative Example 3

[0052] Ultrasonic complexation (2) in Example 1 is omitted, and other conditions or parameters are the same as those in Example 1. Compared with Example 1, after an emulsifier is added, ultrasonic treatment is not used to promote complexation between the amorphous region and the emulsifier, and the final complexation rate and the starch utilization rate are as low as 36% and 41% respectively. Therefore, it can be explained that ultrasonic treatment in step (2) has the effect of promoting complexation between the amorphous region of the starch and the emulsifier.

Comparative Example 4

[0053] Ultrasonic de-clustering (1) in Example 1 is changed into gelatinization: 5%-10% waxy corn starch slurry is prepared, put into a boiling water bath for 40 minutes, stirred while boiling and cooled to 60 C. for heat preservation. Compared with Example 1, the complexation rate and the starch utilization rate are reduced to 34% and 36% respectively.

TABLE-US-00001 TABLE 1 Complexation Starch utilization Sample rate/% rate/% Example 1 65 72 Example 2 58 69 Comparative Example 1 0.75 13 Comparative Example 2 32 37 Comparative Example 3 36 41 Comparative Example 4 34 36

[0054] It can be seen from Table 1 that through ultrasonic de-clustering and ultrasonic induced emulsifier complexation (namely Example 1), the starch amorphous region-emulsifier complexation rate and the starch utilization rate in a water reaction system can reach up to 65% and 72% respectively. Comparative Example 1 is equivalent to a blank control. The waxy corn starch is directly enzymatically hydrolyzed after an emulsifier is added. Because the starch chain is not stretched, it is almost impossible to achieve complexation, and only the surface parts of starch granules can be enzymatically hydrolyzed. Without ultrasonic de-clustering or ultrasonic complexation (Comparative Example 2 and Comparative Example 3), the starch-emulsifier complex rates in the amorphous region are 32% and 36% separately, and the starch utilization rates are 37% and 41% respectively, which are increased by 24% and 28% respectively in comparison with those of the blank group. On the other hand, Comparative Examples 2 to 3 are compared with Example 1, the increase effect (increased by 59%) of the starch utilization rate after combined treatment of ultrasonic de-clustering and ultrasonic complexation is better than the total effect (24%+28%=52%) of separate ultrasonic de-clustering and separate ultrasonic complexation, indicating that ultrasonic de-clustering and ultrasonic complexation support each other in increasing the starch utilization rate and has a certain synergistic effect during combined treatment. Ultrasonic de-clustering (1) in Example 1 is replaced with gelatinization in Comparative Example 4, which is also the common method of preparing amylodextrin currently. In this step, the starch crystallization region is destroyed at a high temperature, the starch is decomposed into many small molecule sugars and other fragments (see Table 2 for details) in the enzymolysis process, and the total amylodextrin yield is greatly reduced; compared with Example 1, the complexation rate and the starch utilization rate are reduced to 34% and 36% respectively, indicating that ultrasonic de-clustering can protect the starch crystallization region when performed at a temperature lower than the starch gelatinization temperature, so as to avoid the situation that the starch crystallization region is degraded into small molecular sugars and other fragments in subsequent enzymolysis, thereby obtaining amylodextrin with uniform molecular weight distribution to increase the complexation rate and the starch utilization rate. It can be explained that through the combined effects of ultrasonic de-clustering and ultrasonic accelerated emulsifier complexation, not only is the destruction of the starch crystallization region avoided, but also the starch amorphous region-emulsifier complex rate and the starch utilization rate are increased.

TABLE-US-00002 TABLE 2 Chain length distribution of amylodextrin Membrane molecular weight cutoff DP < 10 DP10-30 DP > 30 Content (Da) (%) (%) (%) (%) Example 1 <2000 86.7 0.8 12.6 0.4 0.7 0.5 6.1 0.1 2000-5000 0.4 0.3 92.4 0.1 7.2 0.7 81.9 0.3 >5000 16.5 0.4 15.6 0.6 67.9 0.2 12.0 0.8 Example 2 <2000 84.8 0.5 11.9 0.2 3.3 0.6 5.4 0.4 2000-5000 1.8 0.1 91.7 0.3 6.5 0.7 78.7 0.5 >5000 16.5 0.6 13.2 0.7 70.3 0.1 15.9 0.6 Compara- <2000 91.5 0.8 6.4 0.6 2.1 0.3 25.3 0.1 tive 2000-5000 15.6 0.4 80.4 0.8 4.0 0.5 69.8 0.6 Example 4 >5000 24.6 0.7 16.9 0.3 58.5 0.3 4.9 0.7 Note: DP refers to degree of polymerization

[0055] It can be seen from Table 2 that the amylodextrin obtained by complex enzymolysis of the waxy corn starch is divided into three components through membrane separation in Examples 1 and 2, main ingredients of the component with a molecular weight of lower than 2000 Da include small molecular glucose, ethanol, an emulsifier and other substances, and the content is relatively low, only 5.4%-6.1%; the component with a molecular weight higher than 2000 Da is the amylodextrin with high purity, the content of the amylodextrin with the molecular weight cutoff of 2000-5000 Da, i.e., DP10-30 is the highest, accounting for 78.7%-81.9%; the content of the amylodextrin with the molecular weight cutoff higher than 5000 Da, i.e., DP>30 is the second, accounting for 12%-15.9%. It can be explained that the degree of polymerization of the amylodextrin obtained in the disclosure is mostly distributed between 10 and 30. In Comparative Example 4, the starch is gelatinized at a high temperature first. This step destroys the starch crystallization region, and starch molecules are degraded into many small molecule sugars and other fragments during complex enzymolysis. Therefore, the content of components with molecular weight lower than 2000 Da is higher than that in Example 1 and Example 2, accounting for about 25%.