An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan

Abstract

This application relates to the field of dextran, specifically disclosing an industrial-grade continuous preparation process for producing high-purity highland barley β-glucan, comprising the following steps: S1, pulverization; S2, extraction and enzymatic hydrolysis: extraction at 65-85℃ for 2-6 hours, solid-liquid ratio of 1:(12-20), using 0.4-1.0 wt% α-amylase for 2-3 times; S3, alcohol precipitation: precipitating the supernatant obtained in step S2 with a 40-60 wt% ethanol solution; S4, purification: purification at 30-50℃ for 2-12 hours, solid-liquid ratio of 1:(60-90), using 0.15-0.25 wt% α-amylase to obtain highland barley β-glucan. The preparation process of this application yields highland barley β-glucan with a purity of up to 80.35%, achieving a significant improvement in cost reduction and efficiency.

Description

Technical Field

[0001] This application relates to the technical field of dextran, and in particular to an industrial-grade continuous preparation process for producing high-purity highland barley β-glucan. Background Technology

[0002] Barley, a variety of wheat in the Poaceae family, is one of the world's oldest crops, boasting a long history of cultivation. Thousands of years ago, barley was cultivated and utilized by humans. Over its long development, barley became a staple food, with a history of consumption exceeding 3,500 years. Barley is rich in protein, beta-glucan, fiber, vitamins, and other nutrients, containing 18 amino acids, various unsaturated fatty acids, and certain minerals. These nutrients not only give barley high nutritional value but also provide ample room for the development of its derivative products. Since the commencement of barley hybridization breeding in 1965, through the unremitting efforts of breeders, a series of main varieties such as "Zangqing 320" and "Zangqing 2000" have been developed. Simultaneously, products such as barley cordyceps wine and barley rhodiola wine have been developed around barley, focusing on their effects of regulating the immune system and combating fatigue, further promoting the development of the barley industry. β-glucan from highland barley, as an important extract of highland barley, plays a key role in fully realizing the nutritional value of highland barley.

[0003] In existing technologies for preparing β-glucan from highland barley, the extraction process typically involves first adding the raw material to water to allow a reaction, followed by the addition of a specific enzyme for enzymatic hydrolysis. This step-by-step process is relatively complex. Alternatively, under certain conditions, other common enzymes may be used to extract or enzymatically hydrolyze the crushed highland barley separately, followed by further processing of the products. Purification primarily uses ethanol, and in large quantities. Some processes, after extraction and enzymatic hydrolysis, involve multiple separation and concentration steps followed by the addition of a large amount of ethanol to precipitate crude β-glucan extract, which is then further purified. Others involve multi-stage ethanol purification of the crude extract to improve the purity of the β-glucan.

[0004] Existing technologies for preparing β-glucan from highland barley have high costs, making mass production difficult. Even if mass production is achieved, the purity of the resulting β-glucan from highland barley can only reach around 60%, which cannot meet practical requirements. Summary of the Invention

[0005] To address the aforementioned technical problems, this application provides an industrial-grade continuous preparation process for producing high-purity highland barley β-glucan.

[0006] This application provides an industrial-grade continuous preparation process for producing high-purity highland barley β-glucan, comprising the following steps:

[0007] S1. Grinding: Grind the highland barley into powder;

[0008] S2. Extraction and enzymatic hydrolysis: Under the conditions of a temperature of 65-85℃, an extraction time of 2-6h, and a solid-liquid ratio of 1:(12-20), the pulverized highland barley rice in step S1 is extracted and enzymatically hydrolyzed using an α-amylase solution to obtain a mixed solution. The amount of α-amylase in the α-amylase solution is 0.4-1.0wt% of the total amount of pulverized highland barley rice, and the extraction and enzymatic hydrolysis are performed 2-3 times.

[0009] S3. Alcohol precipitation: After the mixture obtained in step S2 is allowed to stand, the supernatant is obtained, filtered, concentrated, and precipitated with an ethanol solution with a concentration of 40-60 wt% to obtain a suspension of crude β-glucan extract. Then, it is filtered to obtain a solid substance, dried, and pulverized to obtain crude β-glucan extract.

[0010] S4. Purification: Under the conditions of a temperature of 30-50℃, a purification time of 2-12h, and a solid-liquid ratio of 1:(60-90), the crude β-glucan extract in step S3 is purified using an α-amylase solution. The extract is filtered to obtain a solid substance, which is then dried and pulverized to obtain barley β-glucan. The amount of α-amylase in the α-amylase solution is 0.15-0.25wt% of the total amount of crude β-glucan extract.

[0011] Preferably, in step S2, the temperature is 75°C.

[0012] Preferably, in step S2, the extraction time is 2 hours.

[0013] Preferably, in step S2, the solid-liquid ratio is 1:14.

[0014] Preferably, in step S2, the amount of α-amylase used is 0.5 wt% of the total amount of crushed highland barley.

[0015] Preferably, in step S3, the concentration of the ethanol solution is 60 wt%.

[0016] Preferably, in step S4, the temperature is 50°C.

[0017] Preferably, in step S4, the purification time is 4 hours.

[0018] Preferably, in step S4, the solid-liquid ratio is 1:80.

[0019] Preferably, in step S4, the amount of α-amylase used is 0.2 wt% of the total amount of the pulverized crude β-glucan extract.

[0020] By adopting the above technical solution, this application first pulverizes highland barley rice to 80 mesh for easy subsequent processing. Then, only α-amylase is used in the extraction and enzymatic hydrolysis process. By strictly setting the temperature, time, solid-liquid ratio, number of times, and dosage of α-amylase, and by carrying out the extraction and enzymatic hydrolysis reactions simultaneously, time is saved while ensuring the quality of extraction and enzymatic hydrolysis. The highland barley β-glucan content in the crude β-glucan extract is still high, resulting in a mixture containing a large amount of crude β-glucan extract. After standing for a period of time, the supernatant is obtained and filtered. Two-stage filtration is performed using plate and frame centrifugation and tubular centrifugation in series, which is low-cost, suitable for mass production, and improves filtration efficiency. Then, the mixture is concentrated to 1 / 3 of the original volume and then ethanol is added to the system until the ethanol concentration reaches a certain value. Alcohol precipitation is then performed to obtain a suspension of crude β-glucan extract, which is then filtered to obtain a solid substance (wet precipitate). This solid substance is then dried in a vacuum environment and pulverized to 50 mesh to obtain the crude β-glucan extract. Subsequently, this application purified the crude β-glucan extract, strictly controlling the temperature, solid-liquid ratio, time, and α-amylase dosage. α-amylase was still used to treat the crude extract. Unlike existing purification methods that use large amounts of ethanol, this application uses α-amylase, significantly reducing the amount of ethanol used in the overall process. After purification, the α-amylase was inactivated by heating. The precipitate was then collected, dried under vacuum, and pulverized to 80 mesh to obtain barley β-glucan.

[0021] In summary, this application has the following beneficial technical effects: the preparation process of this application has low cost, is easy to operate, and is convenient for mass production. Moreover, the purity of the final barley β-glucan can reach up to 80.35%, which has achieved a significant progress in cost reduction and efficiency improvement. It has the dual advantages of large-scale operation and good product quality. Detailed Implementation

[0022] The present application will be further described in detail below with reference to embodiments and comparative examples.

[0023] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan includes the following steps:

[0024] S1. Grinding: Grind the highland barley into powder;

[0025] S2. Extraction and enzymatic hydrolysis: Under the conditions of a temperature of 65-85℃, an extraction time of 2-6h, and a solid-liquid ratio of 1:(12-20), the pulverized highland barley rice in step S1 is extracted and enzymatically hydrolyzed using an α-amylase solution to obtain a mixed solution. The amount of α-amylase in the α-amylase solution is 0.4-1.0wt% of the total amount of pulverized highland barley rice, and the extraction and enzymatic hydrolysis are performed 2-3 times.

[0026] S3. Alcohol precipitation: After the mixture obtained in step S2 is allowed to stand, the supernatant is obtained, filtered, concentrated, and precipitated with an ethanol solution with a concentration of 40-60 wt% to obtain a suspension of crude β-glucan extract. Then, it is filtered to obtain a solid substance, dried, and pulverized to obtain crude β-glucan extract.

[0027] S4. Purification: Under the conditions of a temperature of 30-50℃, a purification time of 2-12h, and a solid-liquid ratio of 1:(60-90), the crude β-glucan extract in step S3 is purified using an α-amylase solution. The extract is filtered to obtain a solid substance, which is then dried and pulverized to obtain barley β-glucan. The amount of α-amylase in the α-amylase solution is 0.15-0.25wt% of the total amount of crude β-glucan extract.

[0028] In a preferred embodiment, the temperature in step S2 is 75°C.

[0029] In a preferred embodiment, the extraction time in step S2 is 2 hours.

[0030] In a preferred embodiment, in step S2, the solid-liquid ratio is 1:14.

[0031] In a preferred embodiment, in step S2, the amount of α-amylase used is 0.5 wt% of the total amount of crushed highland barley.

[0032] In a preferred embodiment, in step S3, the concentration of the ethanol solution is 60 wt%.

[0033] In a preferred embodiment, the temperature in step S4 is 50°C.

[0034] In a preferred embodiment, the purification time in step S4 is 4 hours.

[0035] In a preferred embodiment, in step S4, the solid-liquid ratio is 1:80.

[0036] In a preferred embodiment, in step S4, the amount of α-amylase used is 0.2 wt% of the total amount of the pulverized crude β-glucan extract.

[0037] Example 1

[0038] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan includes the following steps:

[0039] S1. Grinding: Put the barley into a grinder and grind it. Pass it through an 80-mesh sieve and weigh it. Record the total weight as 50kg. Set aside for later use.

[0040] S2. Extraction and enzymatic hydrolysis: Mix all the crushed highland barley rice from step S1, 250g of α-amylase, and 700L of water, and put them into an extraction tank with a capacity of 2 tons. Control the temperature at 75℃ and carry out extraction and enzymatic hydrolysis for 2 hours. Then, centrifuge and filter, collect the filter residue, add water at a solid-liquid ratio of 1:14, and continue extraction and enzymatic hydrolysis for 2 hours to achieve two extractions and enzymatic hydrolysis.

[0041] S3. Alcohol Precipitation: After the mixture obtained in step S2 is allowed to stand, the supernatant is obtained. It is then filtered in two stages using a plate and frame centrifuge and a tubular centrifuge in series. The mixture is concentrated by heating until it reaches 1 / 3 of its original volume. The supernatant is then placed in an explosion-proof alcohol precipitation tank. 95wt% edible ethanol is then slowly added to the tank. Precipitation gradually occurs in the system. When the ethanol concentration in the system reaches 60wt%, a large amount of precipitate has accumulated. After standing, all the precipitate tends to settle to the bottom of the explosion-proof alcohol precipitation tank. Alcohol precipitation is stopped when no more precipitate falls to the bottom of the explosion-proof alcohol precipitation tank, resulting in a suspension of crude β-glucan extract. The suspension is then filtered to collect the wet precipitate, which is then dried in a vacuum environment at 60℃. The solid precipitate is then pulverized in a pulverizer and passed through a 50-mesh sieve to obtain crude β-glucan extract.

[0042] S4. Purification: Take 10 kg of crude β-glucan extract obtained in step S3, mix it with 20 g of α-amylase and 800 L of water, put it into a purification tank with a capacity of 2 tons, control the temperature at 50℃, and purify it for 4 hours. After the purification, use plate and frame centrifugation and tubular centrifugation in series for two-stage filtration, collect the wet precipitate, and obtain solid material. Dry it in a vacuum environment at a temperature of 60℃, put it into a pulverizer for pulverization, and pass it through an 80-mesh sieve to obtain barley β-glucan.

[0043] Example 2

[0044] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, the temperature is controlled at 65°C, while the rest is the same as in Example 1.

[0045] Example 3

[0046] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, the temperature is controlled at 85°C, while the rest is the same as in Example 1.

[0047] Example 4

[0048] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, the extraction time is controlled to be 4 hours, while the rest is the same as in Example 1.

[0049] Example 5

[0050] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, the extraction time is controlled to be 6 hours, while the rest is the same as in Example 1.

[0051] Example 6

[0052] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice, 250g of α-amylase, and 600L of water from step S1 are mixed together, and the solid-liquid ratio is controlled to be 1:12. The rest is the same as in Example 1.

[0053] Example 7

[0054] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice, 250g of α-amylase, and 800L of water from step S1 are mixed together, and the solid-liquid ratio is controlled to be 1:16. The rest is the same as in Example 1.

[0055] Example 8

[0056] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice, 250g of α-amylase, and 900L of water from step S1 are mixed together, and the solid-liquid ratio is controlled to be 1:18. The rest is the same as in Example 1.

[0057] Example 9

[0058] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice, 250g of α-amylase, and 1000L of water from step S1 are mixed together, and the solid-liquid ratio is controlled to be 1:20. The rest is the same as in Example 1.

[0059] Example 10

[0060] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice from step S1, 200g of α-amylase, and 700L of water are mixed together, and the amount of α-amylase is controlled to be 0.4wt% of the total amount of crushed highland barley rice. The rest is the same as in Example 1.

[0061] Example 11

[0062] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice from step S1, 400g of α-amylase, and 700L of water are mixed together, and the amount of α-amylase is controlled to be 0.8wt% of the total amount of crushed highland barley rice. The rest is the same as in Example 1.

[0063] Example 12

[0064] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S2, all the crushed highland barley rice from step S1, 500g of α-amylase, and 700L of water are mixed together, and the amount of α-amylase is controlled to be 1.0wt% of the total amount of crushed highland barley rice. The rest is the same as in Example 1.

[0065] Example 13

[0066] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S3, 95wt% edible ethanol is added to the tank until the ethanol concentration in the system is 40wt%, while the rest is the same as in Example 1.

[0067] Example 14

[0068] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, the temperature is controlled at 30°C, while the rest is the same as in Example 1.

[0069] Example 15

[0070] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, the temperature is controlled at 40°C, while the rest is the same as in Example 1.

[0071] Example 16

[0072] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, the purification time is controlled to be 2 hours, while the rest are the same as in Example 1.

[0073] Example 17

[0074] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, the purification time is controlled to be 6 hours, while the rest are the same as in Example 1.

[0075] Example 18

[0076] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, the purification time is controlled to be 8 hours, while the rest are the same as in Example 1.

[0077] Example 19

[0078] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, the purification time is controlled at 12 hours, while the rest is the same as in Example 1.

[0079] Example 20

[0080] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, 10 kg of crude β-glucan extract obtained in step S3 is taken and mixed with 20 g of α-amylase and 600 L of water, with the solid-liquid ratio controlled at 1:60. The rest is the same as in Example 1.

[0081] Example 21

[0082] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, 10 kg of crude β-glucan extract obtained in step S3 is taken and mixed with 20 g of α-amylase and 700 L of water, with the solid-liquid ratio controlled at 1:70. The rest is the same as in Example 1.

[0083] Example 22

[0084] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, 10 kg of crude β-glucan extract obtained in step S3 is taken and mixed with 20 g of α-amylase and 900 L of water, with the solid-liquid ratio controlled at 1:90. The rest is the same as in Example 1.

[0085] Example 23

[0086] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, 10 kg of the crude β-glucan extract obtained in step S3 is mixed with 15 g of α-amylase and 800 L of water, and the amount of α-amylase is controlled to be 0.15 wt% of the total amount of the pulverized crude β-glucan extract. The rest is the same as in Example 1.

[0087] Example 24

[0088] An industrial-grade continuous preparation process for producing high-purity highland barley β-glucan differs from Example 1 in that: in step S4, 10 kg of the crude β-glucan extract obtained in step S3 is mixed with 25 g of α-amylase and 800 L of water, and the amount of α-amylase is controlled to be 0.25 wt% of the total amount of the pulverized crude β-glucan extract. The rest is the same as in Example 1.

[0089] Comparative Example 1

[0090] 50 kg of highland barley rice was extracted by reflux with 80 wt% ethanol solution (solid-liquid volume ratio 1:15) for 2 h. The precipitate was collected, dried, and the pH was adjusted to 8. The mixture was then extracted by stirring at 65 °C for 2 h. After centrifugation at 4000 r / min for 15 min, the supernatant was adjusted to pH 4.5 with 2 mol / L HCl solution. α-amylase was added at a solid-liquid volume ratio of 1:10, and the mixture was centrifuged at 4000 r / min for 20 min. The temperature of the supernatant was adjusted to 60 °C, and 95 wt% ethanol was added at a volume ratio of 1:2 under a pressure of 0.09 MPa. The mixture was then centrifuged at 4000 r / min for 20 min to obtain highland barley β-glucan.

[0091] Comparative Example 2

[0092] 50 kg of highland barley rice was pretreated with a 75 wt% ethanol solution (solid-liquid volume ratio 1:15), then extracted twice with hot water at 80℃, precipitated with alcohol, and freeze-dried to obtain highland barley β-glucan.

[0093] Comparative Example 3

[0094] Barley grains were cleaned, washed, crushed, and passed through a 40-mesh sieve. They were then gelatinized in a water bath at a solid-liquid ratio of 1:6 and a temperature of 90℃ for 2 hours. After 2 hours, α-amylase was added for enzymatic hydrolysis for 30 minutes. The mixture was then filtered through a 40-mesh sieve to obtain the filtrate. The residue was gelatinized again in a water bath at a solid-liquid ratio of 1:3 and a temperature of 90℃ for 2 hours. The residue was then filtered through a 40-mesh sieve to obtain the second filtrate. The residue was then gelatinized again in a water bath at a solid-liquid ratio of 1:2 and a temperature of 90℃ for 2 hours. The residue was then filtered through a 40-mesh sieve to obtain the third filtrate. The filtrates were combined and centrifuged at 4000 rpm for 10 minutes to obtain the supernatant. 10% hydrogen peroxide was added for decolorization. After complete decolorization, two volumes of ethanol were added for precipitation. The mixture was left overnight and filtered again through a 40-mesh sieve. The precipitate was then centrifuged at 4000 rpm for 10 minutes to obtain the precipitate. The precipitate was then vacuum dried with silica to obtain barley β-glucan.

[0095] Performance testing

[0096] The β-glucan content of the product (crude β-glucan extract) obtained in step S3 and the final product (barley β-glucan) obtained in step S4 in the embodiments and comparative examples of this application were detected and recorded as the enzymatic extraction content and the purified content, respectively. The detection kit used was the Megazyme hybrid β-glucan detection kit, and the results are recorded in Table 1.

[0097] Table 1 Data Record Table

[0098]

[0099] Data Analysis:

[0100] As shown in Table 1, the enzymatic extraction yield of β-glucan in Example 1 reached 45.89%, and the purified content reached 80.35%. In contrast, the purified content in Comparative Example 1 was 74.31%, and the process also used HCl solution, making it inconvenient and relatively complex. The purified content in Comparative Example 2 was 76.94%, and the purified content in Comparative Example 3 was extremely low, and it also included a gelatinization step, making the process complex. Therefore, the preparation process of this application has lower costs, is simpler to operate, and is convenient for mass production. Furthermore, the purity of the final barley β-glucan obtained can reach up to 80.35%, achieving a significant improvement in cost reduction and efficiency, and possessing the dual advantages of large-scale operation and good product quality.

[0101] The embodiments described in this specific implementation are preferred embodiments of this application and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, shape and principle of this application should be covered within the scope of protection of this application.

Claims

1. An industrial grade continuous production process for producing high purity highland barley β-glucan, characterized in that, The method comprises the following steps: S1, crushing: crushing highland barley; S2, extraction and enzymolysis: under the conditions of a temperature of 65-85℃, an extraction time of 2-6h, and a solid-liquid ratio of 1: (12-20), an α-amylase solution is used for extraction and enzymolysis of the crushed highland barley in step S1, to obtain a mixed solution, the α-amylase in the α-amylase solution is used in an amount of 0.4-1.0wt% of the total amount of the crushed highland barley, and the extraction and enzymolysis is performed for 2-3 times; S3, alcohol precipitation: after the mixed solution obtained in step S2 is allowed to stand, a supernatant is obtained, filtered, concentrated, and subjected to alcohol precipitation with an ethanol solution having a concentration of 40-60wt%, to obtain a β-glucan crude extract suspension, which is then filtered to obtain solid substances, dried, crushed, and ground to obtain a β-glucan crude extract; S4, purification: under the conditions of a temperature of 30-50℃, a purification time of 2-12h, and a solid-liquid ratio of 1: (60-90), an α-amylase solution is used for purification of the β-glucan crude extract in step S3, filtered to obtain solid substances, dried in a vacuum environment at a temperature of 60℃, and crushed to obtain highland barley β-glucan, the α-amylase in the α-amylase solution is used in an amount of 0.15-0.25wt% of the total amount of the β-glucan crude extract.

2. The industrial continuous production process for producing high-purity highland barley β-glucan according to claim 1, characterized in that, In step S2, the temperature is 75℃.

3. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S2, the extraction time is 2h.

4. The industrial continuous production process for producing high-purity highland barley β-glucan according to claim 1, characterized in that, In step S2, the solid-liquid ratio is 1:

14.

5. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S2, the α-amylase is used in an amount of 0.5wt% of the total amount of the crushed highland barley.

6. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S3, the concentration of the ethanol solution is 60wt%.

7. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S4, the temperature is 50℃.

8. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S4, the purification time is 4h.

9. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S4, the solid-liquid ratio is 1:

80.

10. The industrial continuous production process for producing high purity highland barley β-glucan according to claim 1, characterized in that, In step S4, the α-amylase is used in an amount of 0.2wt% of the total amount of the crushed β-glucan crude extract.

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