A kind of copper algae polysaccharide enzymatic hydrolyzate and its application
A copper algae polysaccharide and enzymatic hydrolyzate technology, which is applied in the field of seaweed polysaccharide extract, can solve the problems of ineffectiveness and achieve the effects of mild process conditions, lower intestinal pH, and change the structure of intestinal flora
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Embodiment 1
[0031] (1) Enzymatic hydrolysis pretreatment:
[0032] Wash the copper algae, control the water, dry in an oven at 60°C, and pulverize. Choose the copper algae that crosses 100 mesh sieves after the crushing, get 100.0g powder, add in the water of 1500g, adjust pH to be 5.5, add 0.5g compound enzyme, compound enzyme is papain (Shanghai Blue Season Science and Technology Development Co., Ltd., 1,000,000 U / g) mixed with cellulase (Zhangjiagang Jinyuan Biochemical Co., Ltd., 80,000 U / g) in a mass ratio of 1:1, placed in a constant temperature water bath at 50°C, stirred at a constant speed for 60 minutes, centrifuged at 4000rpm for 10 minutes, and taken The clear liquid is the enzymatic hydrolysis pretreatment liquid;
[0033] (2) Alcohol precipitation:
[0034] The enzymatic hydrolysis pretreatment solution was concentrated to 1 L by rotary evaporation, and 95% ethanol solution was added until the final volume fraction of ethanol was 60%, and stood overnight. The sample solu...
Embodiment 2
[0045] The reaction steps and conditions are the same as those in Example 1, the only difference being that in step (4) the reaction was carried out at 50° C. for 3 hours, and finally the enzymatic hydrolysis product SHPA-3 was obtained with an average molecular weight of 10.54 KDa.
Embodiment 3
[0047] Take MRS-glucose medium and 3 MRS sugar-free medium, add SHPA-2, SHPA-3 and FOS (fructo-oligosaccharide) equivalent to glucose respectively in the 3 MRS sugar-free medium (Shanghai Emmons Chemical Technology Co., Ltd.), denoted as MRS-SHPA-2, MRS-SHPA-3, MRS-FOS.
[0048] The fresh feces of the subjects were collected in sterile blue cap bottles and sent for inspection immediately. Quantitatively weigh the specimen, suspend it in ice-cold PBS buffer (pH7.3) to make the fecal sample concentration 50 mg / mL, add sterile glass beads and vortex for 1 min, and let it stand for later use.
[0049] Inoculate the prepared feces suspension into MRS-SHPA-2, MRS-SHPA-3, glucose, and MRS-FOS culture fluid in an amount of 5%, culture on a shaker at 37°C (50rpm), and measure the concentration of the culture fluid after 24h. pH value, see figure 1 . The fermentation broth was serially diluted with PBS buffer solution (pH7.3), and the gradient was set to 10 3 -10 6 Take different g...
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