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Method for extracting auricularia auricular polysaccharide by using biological enzyme method

A black fungus polysaccharide and bio-enzyme technology, which is applied in the field of enzymatic extraction of black fungus polysaccharides, can solve the problems that affect the sustainable development of the industry and the difference in wall breaking effect, and achieve the effect of improving biological activity and good sensory properties

Active Publication Date: 2021-01-29
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2017, the total output of black fungus in my country reached 6.3884 million tons, but only about 6% of the output was used for deep processing of products, which affected the sustainable development of the industry
Due to the different breaking sites of different types of enzymes, there are some differences in the breaking effect

Method used

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  • Method for extracting auricularia auricular polysaccharide by using biological enzyme method
  • Method for extracting auricularia auricular polysaccharide by using biological enzyme method
  • Method for extracting auricularia auricular polysaccharide by using biological enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1 Materials and reagents

[0049] Black fungus (produced in Xiaoxing'an Mountains, Yanjun Farm, Luobei County, Heilongjiang Province). Before use, place the black fungus powder in a 50°C oven for drying, pulverize, and screen the black fungus powder with a mesh number of 150-200 mesh for future use.

[0050] Phenol, concentrated sulfuric acid, sodium hydroxide, ethanol, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, potassium persulfate, chloroform, n-butanol, acetone, ferrous sulfate, ferrous chloride, ferric chloride , potassium ferricyanide, ethylenediaminetetraacetic acid (EDTA), glucose, 1,1-diphenyl-2-picryhydrazyl (DPPH), hydrogen peroxide, phenanthrenolazine (Ferrozine), Nile red fluorescent dye, ascorbic acid (Vc), atorvastatin calcium, 2,2-azino-bis(3-ethyl-benzothiazoline sulfonic acid-6) ammonium salt (ABTS), and the water used in the experiment was deionized water.

[0051] N2 wild-type Caenorhabditis elegans and...

Embodiment 2

[0201] Black fungus pretreatment: Take 10g of black fungus powder (passed through a 150-200 mesh sieve) and add it to 600mL of deionized water, swell at 25°C for 12h, then freeze in a -20°C refrigerator for 60min, and thaw in a constant temperature water bath at 20°C for 3 times Freeze-thaw cycle pretreatment, and then freeze-drying (initial temperature is -30 ° C, vacuum degree is 80 Pa), to obtain 10 g of pretreated black fungus powder.

[0202] Black fungus polysaccharide extraction: take 10g of pretreated black fungus powder, add 600mL deionized water, then add 0.4g cellulase and 0.6g papain (that is, half of the optimal enzyme amount), and Under the conditions of 55°C, pH 5.3, and solid-liquid ratio of 1g / 60mL, act for 3.0h to fully and effectively break the cell wall of black fungus; add 0.6g of mild-acting β-glucanase according to E / S 6.0%, and under other conditions Under the same conditions (55°C, pH 5.3), act for 2 hours. Then heat to boil for 15min to inactivate th...

Embodiment 3

[0234] Accurately weigh 20 g of black fungus powder (150-200 mesh sieve) and dissolve in 560 mL of deionized water, swell for 16 h, then freeze in a -20 °C refrigerator for 50 min, thaw in a constant temperature water bath at 20 °C, perform 4 freeze-thaw cycles, and then After freeze-drying (the initial temperature is -30° C., and the vacuum degree is 80 Pa), 20 g of pretreated black fungus powder is obtained.

[0235] Add 20g of black fungus powder after pretreatment, add 0.8g cellulase and 1.2g papain (that is, according to half of the optimal enzyme amount) and 1200mL deionized water, and then at an action temperature of 45°C, pH 4.0, Act for 1.0h under the condition of ratio 1 / 60 to fully and effectively break the cell wall of black fungus; then add 0.8g of mild-acting β-glucanase according to E / S 4.0%, and act at 45°C and pH 4.0 2h. Prepare 6.54g of black fungus polysaccharides with the method of Example 2, the average polysaccharide yield is 35.06%, ABTS + The scavengi...

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Abstract

The invention discloses a method for extracting auricularia auricular polysaccharide by using a biological enzyme method, which comprises the following steps: adding auricularia auricular powder intodeionized water, adding cellulase and papain, and extracting for 1-5h at 45-80 DEG C under the condition that the pH value is 4.0-8.0; adding one of beta glucanase or mannase or neutral protease, andextracting for 1-5h under the conditions that the temperature is 45-80 DEG C and the pH value is 2.0-9.0; then heating to deactivate enzyme, centrifuging, taking supernatant, adding 95% ethanol, standing for 10 hours at 4 DEG C, and centrifuging; taking the precipitate, dissolving the precipitate in deionized water, deproteinizing by a sevag method until the upper layer has no absorption peak at 280nm, concentrating, and freeze-drying to obtain the auricularia auricular polysaccharide. The extraction yield of the auricularia auricula polysaccharide reaches 36.43%, the removal rate of ABTS < +> is 98.32%, the removal rate of DPPH free radicals is 92.74%, and the removal rate of hydrogen peroxide is 66.56%.

Description

[0001] (1) Technical field [0002] The invention relates to an enzymatic extraction method of black fungus polysaccharide. [0003] (2) Background technology [0004] Black fungus (Auricularia auricula) is an important medicinal and edible colloid fungus in my country. More than 60% of its fruiting body dry weight is polysaccharides, which have multiple functions such as lowering blood sugar, lowering blood fat, immune regulation, and anti-oxidation. In 2017, the total output of black fungus in my country reached 6.3884 million tons, but only about 6% of the output was used for deep processing of products, which affected the sustainable development of the industry. The main reason restricting the extraction and further processing of black fungus polysaccharides is that the cell wall of black fungus is different from the plant cell wall, and the multi-layer compact and complex network cell wall structure makes it extremely complex and tough. [0005] At present, the traditiona...

Claims

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Application Information

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IPC IPC(8): C08B37/00
CPCC08B37/0003
Inventor 张拥军黄齐齐陈双肖斌王为民
Owner CHINA JILIANG UNIV
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