阅读说明：本技术 一种提高糖液中一糖含量的生产工艺 (Production process for increasing content of monosaccharide in sugar solution ) 是由 孙钦波 侍述志 张婷婷 刘世周 刘庆波 赵杰 于 2021-10-18 设计创作，主要内容包括：本发明属于生物技术领域,公开了一种提高糖液中一糖含量的生产工艺,其包括如下步骤：取DE值为20%的淀粉液化液,调节pH为4-5,维持温度为60℃,添加葡萄糖淀粉酶制剂,用量为200U/g淀粉,搅拌反应30h,得到糖化液。本发明通过对酶固定化处理,提高了酶的稳定性,节约酶制剂使用量,提高了葡萄糖含量。(The invention belongs to the technical field of biology, and discloses a production process for increasing the content of monosaccharide in sugar liquid, which comprises the following steps: taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4-5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid. The invention improves the stability of enzyme, saves the using amount of enzyme preparation and improves the glucose content by carrying out enzyme immobilization treatment.)

1. A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

taking starch liquefaction liquid, adjusting pH to 4-5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

2. The process of claim 1, wherein the glucoamylase preparation is prepared according to the following process:

mixing locust bean gum and gelatin, adding into a container containing purified water, heating to 85 deg.C, stirring for dissolving, adding chitosan and sodium alginate under heat preservation, mixing, rapidly defoaming with ultrasonic wave until no bubble is formed, dripping into calcium chloride solution, curing at 4 deg.C for 1-5 hr, washing off surface calcium chloride, adsorbing in glucoamylase for 0.5-2 hr, drying at 20-30 deg.C for 30-50min in a vacuum low-temperature dryer to obtain granule microsphere, and storing at O deg.C to obtain glucoamylase preparation.

3. The process according to claim 1, wherein the starch liquefaction liquor has a DE of 20%.

4. The process of claim 1, wherein the starch liquefaction liquor is prepared by enzymatic hydrolysis of corn starch with amylase.

5. The production process according to claim 2, wherein the mass ratio of the locust bean gum, the gelatin, the chitosan, the sodium alginate and the purified water is 4:2:1:5: 300.

6. The process according to claim 2, wherein the calcium chloride solution is a calcium chloride solution

The concentration of (2) is 50 g/L.

7. The production process according to claim 2, characterized in that the curing time is 2 h.

8. The production process according to claim 2, characterized in that the adsorption time is 1 h.

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a production process for increasing the content of monosaccharide in sugar liquid.

Background

Glucose is one of the most commonly used medical agents and is also an industrial raw material in the fine chemical industry, food industry, fermentation industry and the like. It is well known that glucose is one of the most important downstream products of starch and is also an important further processed product of corn. In the United states, sucrose has been replaced comprehensively by starch sugar produced from corn as a raw material, and the starch sugar is widely used in various fields such as food processing and household eating. The corn has abundant corn resources, and meanwhile, the corn is easy to transport and store, is not limited by seasons, has high starch content, multiple byproduct types and high utilization value, so the corn gradually becomes a main raw material for producing the starch in China, and the corn starch accounts for about 90 percent of the total starch yield in China at present. Therefore, the preparation of glucose by using corn starch becomes an ideal choice for preparing glucose.

An enzyme method is generally used for converting starch into glucose, but free enzyme has poor stability in industrial application, is easily influenced by external factors and is denatured and inactivated; and after the reaction is finished, even if the enzyme has high activity, the enzyme is difficult to recycle, and simultaneously, the enzyme and the product are mixed together after the reaction, which brings certain difficulty to the separation and purification of the product, thereby limiting the industrial application of the tyrosinase to a certain extent. The immobilization of the enzyme can solve the problems, and compared with free enzyme, the immobilized enzyme can repeatedly carry out batch reaction and column-packed continuous reaction for a longer time, so that the use efficiency of the enzyme can be improved, the yield of the product can be increased, and the production cost can be reduced; after the reaction is finished, the immobilized enzyme is easily separated from a substrate and a product, the purification process is simplified, and the quality of the product is improved, so that more and more researches on the immobilization of the enzyme are carried out at home and abroad, but the immobilized enzyme which is really put into industrial application is few, and the main reasons are high immobilization cost, low efficiency and poor stability.

The enzyme immobilization methods reported at present mainly include adsorption, embedding, crosslinking and the like. The enzyme may be immobilized by various immobilization methods, but any one of the immobilization carrier and the immobilization method cannot be applied to all enzymes. For different enzymes, in order to obtain a better immobilization effect and prepare an immobilized enzyme with optimal performance, a proper immobilization method and a proper carrier are selected to improve the activity and the stability of the enzyme according to specific enzymes, substrates and catalytic reaction types, and specifically, which immobilization method is adopted is still based on the property of the enzyme, has no ready rule and can be circulated, only depends on the accumulation of experimental experience, and finally, the evaluation standard of whether the activity of the enzyme can be furthest reserved and the stability of the enzyme can be improved is taken as an evaluation standard.

Disclosure of Invention

In order to overcome the technical defects of the enzyme preparation in the prior art and improve the yield of glucose to the maximum extent, the invention provides a production process for improving the content of monosaccharide in sugar liquid.

In order to realize the purpose of the invention, the following technical scheme is adopted:

a production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

taking starch liquefaction liquid, adjusting pH to 4-5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Further, the glucoamylase preparation is prepared according to the following process:

mixing locust bean gum and gelatin, adding into a container containing purified water, heating to 85 deg.C, stirring for dissolving, adding chitosan and sodium alginate under heat preservation, mixing, rapidly defoaming with ultrasonic wave until no bubble is formed, dripping into calcium chloride solution, curing at 4 deg.C for 1-5 hr, washing off surface calcium chloride, adsorbing in glucoamylase for 0.5-2 hr, drying at 20-30 deg.C for 30-50min in a vacuum low-temperature dryer to obtain granule microsphere, and storing at O deg.C to obtain glucoamylase preparation.

Preferably, the DE value of the starch liquefaction liquor is 20%.

Preferably, the starch liquefaction liquid is prepared by enzymolysis of corn starch by amylase.

Preferably, the mass ratio of the locust bean gum to the gelatin to the chitosan to the sodium alginate to the purified water is 4:2:1:5: 300.

Preferably, the concentration of the calcium chloride solution is 50 g/L.

Preferably, the curing time is 2 h.

Preferably, the adsorption time is 1 h.

The beneficial effects of the invention mainly comprise:

the invention improves the stability of enzyme, saves the using amount of enzyme preparation and improves the glucose content by carrying out enzyme immobilization treatment.

In the process of immobilizing the enzyme, the chitosan has stable chemical property and good heat resistance, and amino groups existing in molecules of the chitosan are easy to be covalently bonded with the enzyme and can be complexed with metal ions, so that the enzyme is prevented from being inhibited by the metal ions and is easy to modify through grafting.

Locust bean gum interacts with gelatin to form a complex in solution, so that the effect of the gel is enhanced; the sodium alginate and the composite colloid are cooperatively immobilized to prepare the microcapsule, and compared with the method for preparing the microcapsule by singly utilizing the sodium alginate, the method has the advantages of providing larger immobilization space and being more beneficial to the diffusion of internal and external substances.

The interaction of the locust bean gum and the gelatin is obviously superior to the compatibility of other colloids, and the chitosan is combined, so that the stability and the catalytic efficiency of the enzyme are greatly improved.

Drawings

FIG. 1: the DE values of different groups are compared.

Detailed Description

Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the products and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations and modifications, or appropriate alterations and combinations, of the products and methods described herein may be made and utilized without departing from the spirit, scope, and spirit of the invention. For a further understanding of the present invention, reference will now be made in detail to the following examples.

Example 1

A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

mixing locust bean gum and gelatin, adding into a container containing purified water, heating to 85 ℃, stirring for dissolving, adding chitosan and sodium alginate under the condition of heat preservation, uniformly mixing, quickly defoaming by ultrasonic until no bubbles exist, then dripping into 50g/L calcium chloride solution with twice volume, curing for 2h at 4 ℃, washing off calcium chloride on the surface, then placing into glucoamylase (10 ten thousand U/ml) for adsorption for 1h, then placing into a vacuum low-temperature dryer for drying for 50min at 20 ℃ to obtain particle microspheres, and storing at O ℃ to obtain the glucoamylase preparation.

Taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4.5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Comparative example 1

A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

mixing locust bean gum and gelatin, adding into a container containing purified water, heating to 85 ℃, stirring for dissolving, adding sodium alginate under the condition of heat preservation, wherein the mass ratio of the locust bean gum to the gelatin to the sodium alginate to the purified water is 4:2:5:300, uniformly mixing, quickly defoaming by ultrasonic waves until no bubbles exist, then dripping into 50g/L calcium chloride solution with twice volume, curing for 2h at 4 ℃, washing off calcium chloride on the surface, then placing into glucoamylase (10 ten thousand U/ml) for adsorption for 1h, then placing into a vacuum low-temperature dryer for drying for 50min at 20 ℃ to obtain particle microspheres, and storing at O ℃ to obtain the glucoamylase preparation.

Taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4.5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Comparative example 2

A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

mixing agar and gelatin, adding into a container containing purified water, heating to 85 ℃, stirring for dissolving, adding chitosan and sodium alginate under the condition of heat preservation, wherein the mass ratio of the agar to the gelatin to the purified water is 4:2:1:5:300, uniformly mixing, quickly defoaming by ultrasonic waves until no bubbles exist, then dripping into 50g/L calcium chloride solution with twice volume, solidifying for 2h at 4 ℃, washing off calcium chloride on the surface, then placing into glucoamylase (10 ten thousand U/ml) for adsorption for 1h, then placing into a vacuum low-temperature dryer for drying for 50min at 20 ℃ to obtain granular microspheres, and storing at O ℃ to obtain the glucoamylase preparation.

Taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4.5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Comparative example 3

A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

mixing locust bean gum and carrageenan, adding the mixture into a container containing purified water, heating the mixture to 85 ℃, stirring and dissolving, adding chitosan and sodium alginate under the condition of heat preservation, uniformly mixing, quickly defoaming the mixture by ultrasonic waves until no bubbles exist, dripping the mixture into 50g/L calcium chloride solution with twice volume, solidifying the mixture for 2 hours at 4 ℃, washing off calcium chloride on the surface, adsorbing the mixture in glucoamylase (10 ten thousand U/ml) for 1 hour, drying the mixture in a vacuum low-temperature dryer for 50 minutes at 20 ℃ to obtain particle microspheres, and storing the microspheres at O ℃, thereby preparing the glucoamylase preparation.

Taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4.5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Comparative example 4

A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

adding locust bean gum into a container containing purified water, heating to 85 ℃, stirring for dissolving, adding chitosan and sodium alginate under the condition of heat preservation, wherein the mass ratio of the locust bean gum to the chitosan to the sodium alginate to the purified water is 6:1:5:300, uniformly mixing, quickly defoaming by ultrasonic waves until no bubbles exist, then dripping the mixture into 50g/L calcium chloride solution with twice volume, curing for 2 hours at 4 ℃, washing off calcium chloride on the surface, then placing the mixture into glucoamylase (10 ten thousand U/ml) for adsorption for 1 hour, then placing the mixture into a vacuum low-temperature dryer for drying for 50 minutes at 20 ℃ to obtain particle microspheres, and storing the particle microspheres at O ℃ to obtain the glucoamylase preparation.

Taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4.5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Comparative example 5

A production process for increasing the content of monosaccharide in sugar liquid comprises the following steps:

adding gelatin into a container containing purified water, heating to 85 ℃, stirring for dissolving, adding chitosan and sodium alginate under the condition of heat preservation, wherein the mass ratio of the gelatin to the purified water to the chitosan to the sodium alginate to the purified water is 6:1:5:300, uniformly mixing, quickly defoaming by ultrasonic waves until no bubbles exist, then dropwise adding the mixture into a 50g/L calcium chloride solution, curing for 2 hours at 4 ℃, washing off calcium chloride on the surface, then placing the mixture into glucoamylase (10 ten thousand U/ml) for adsorption for 1 hour, then placing the mixture into a vacuum low-temperature dryer for drying for 50 minutes at 20 ℃ to obtain granular microspheres, and storing the granular microspheres at O ℃ to obtain the glucoamylase preparation.

Taking starch liquefaction liquid with DE value of 20%, adjusting pH to 4.5, maintaining temperature at 60 deg.C, adding glucoamylase preparation with dosage of 200U/g starch, stirring and reacting for 30 hr to obtain saccharified liquid.

Example 2

DE values of the saccharified liquids of different test examples were compared.

The DE values of example 1 and the respective proportions were determined by setting the saccharification reaction times at 25,30,35,40,45, 50 in h (the percentage of glucose equivalents in dry matter is referred to as DE value). As shown in FIG. 1, the DE value of the example 1 group approaches the peak value at 30h as the enzymolysis time increases, while the free enzyme group and the comparative example 4 group need longer enzymolysis time, and compared longitudinally, the DE value of the example 1 group is the highest, and compared with the comparative example 1 group, the DE value is the lowest and is the free enzyme group.

Example 3

The enzyme activity preservation rates of the enzyme preparations of different experimental examples are compared.

Under the same conditions, the enzyme activity preservation rates of the respective test examples were compared. The first enzyme activity is defined as 100%, and the preservation rate of the subsequent reaction enzyme activity refers to the preservation rate relative to the first enzyme activity.

TABLE 1

Group of	Preservation Rate of 2 nd reaction%	The preservation ratio of the 3 rd reaction%	Preservation Rate of 4 th reaction%
				Example 1	96.4	88.7	83.1
Comparative example 1	93.9	85.2	80.8
				Comparative example 2	91.3	82.5	77
Comparative example 3	92.6	81.4	73.2
				Comparative example 4	85.1	71.7	60.3
Comparative example 5	80.6	68.5	61.9

As shown in Table 1, the enzymes all have certain reaction stability after being immobilized, but different immobilization methods have great difference, the preservation rate of the enzyme activity of example 1 is the highest, the preservation rate of the 2 nd reaction reaches more than 96%, and the preservation rate of the 4 th reaction is still maintained at 83%.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.