阅读说明：本技术 一种高效提取玛咖多糖的方法 (Method for efficiently extracting maca polysaccharide ) 是由 李爱民 吴晓磊 李永强 李颖 邢岩 于 2020-06-29 设计创作，主要内容包括：一种高效提取玛咖多糖的方法,该方法将玛咖粉碎过筛,经超声波辅助热水浸提后,过滤得到的上清液,经过酶解去淀粉,Sevage法去除蛋白,冻干后得到玛咖多糖。该方法操作简单、提取温度低时间短、提取率高、纯度高,获得的三个多糖成分具有较好的抗氧化性和降血脂功效。(A method for efficiently extracting maca polysaccharide comprises the steps of crushing and sieving maca, carrying out ultrasonic-assisted hot water extraction, filtering to obtain supernatant, carrying out enzymolysis to remove starch, carrying out Sevage method to remove protein, and carrying out freeze-drying to obtain the maca polysaccharide. The method has the advantages of simple operation, low extraction temperature, short extraction time, high extraction rate, and high purity, and the obtained three polysaccharide components have good oxidation resistance and blood lipid reducing effect.)

1. A method for efficiently extracting maca polysaccharide is characterized by comprising the following steps:

I. crushing and sieving maca roots and stems, and extracting water-soluble substances in maca powder by adopting an ultrasonic-assisted hot water extraction method;

II, filtering the mixture obtained in the step I to obtain a maca water extract, carrying out enzymolysis by sequentially adopting amylase and glucoamylase to remove starch, then inactivating the enzymes, and centrifuging to obtain a supernatant;

III, adding ethanol into the supernatant obtained in the step II, and standing at a low temperature to precipitate polysaccharide;

and IV, re-dissolving the precipitate obtained in the step III with water, removing protein from the re-dissolved solution by adopting a Sevage method, and freeze-drying to obtain the maca crude polysaccharide.

2. The method of claim 1, wherein the specific operation of step I is: the extraction temperature is 40-60 ℃, the extraction time is 30-60min, the ultrasonic power is 150-300W, and the material-liquid ratio (weight/volume) is 1: 20-40 g/mL.

3. The method according to claim 1, wherein the specific operation of step II is: adding amylase into a maca water extract, wherein the mass ratio of the amylase to the raw maca powder is 1: 100-2000, and reacting for 2-5 hours at 30-80 ℃; and adding saccharifying enzyme, wherein the mass ratio of the saccharifying enzyme to the raw material maca powder is 1: 50-1000, reacting for 1-4 h at 30-80 ℃, then heating at 90-100 ℃ for 5-30 min to inactivate the enzyme, and centrifuging to obtain supernatant.

4. The method according to claim 3, wherein the specific operation of step II is: adding amylase into a maca water extract, wherein the mass ratio of the amylase to the raw maca powder is 1: 1000; reacting at 60 ℃ for 3h, adding saccharifying enzyme, reacting at 60 ℃ for 2h, heating at 100 ℃ for 10min to inactivate enzyme, and centrifuging to obtain supernatant.

5. The method of claim 1, wherein the amylase of step II is an alpha amylase.

6. The method of claim 1, wherein in step III, the polysaccharide solution and ethanol are placed at 4-10 ℃ for 10-14 h at a volume ratio of 1: 2-4, so that the polysaccharide precipitates and separates.

7. The method of claim 1, wherein the specific operation of step IV is: preparing a Sevage reagent, wherein the volume ratio of n-butyl alcohol to chloroform is 1: 2-5; mixing and shaking the Sevage reagent and the polysaccharide solution according to the volume fraction of 1: 1-4, centrifuging and layering, reserving the upper layer solution, and then adding the Sevage reagent again for repeated operation until no protein is detected in the upper layer solution.

8. The method of any one of claims 1-7, further comprising:

step V: and (3) separating the maca crude polysaccharide obtained in the step (IV) by using an anion exchange column to obtain three polysaccharide components, namely MPS1, MPS2 and MPS 3.

9. The method of claim 8, wherein the anion exchange column separation conditions are:

the ion exchange column adopts polymethyl methacrylate resin as the filler and ddH as the solvent₂O, 0.1M NaCl and 0.2M NaCl; ddH₂O elution gave MPS1, 0.1M NaCl elution gave MPS2, and 0.2M NaCl elution gave MPS 3.

10. The method according to claim 9, wherein the MPS1 has a monosaccharide composition and mass ratio of arabinose galactose glucose mannose 1.9: 1.4: 3.8: 1; the monosaccharide composition and the mass ratio of MPS2 are arabinose, galactose and glucose which are 1.8: 1.3: 1; the monosaccharide composition and mass ratio of MPS3 are rhamnose, arabinose, galactose and glucose 1: 15.6: 24.3: 5.3.

Technical Field

The invention belongs to the technical field of extraction of effective components of plants, and particularly relates to a method for efficiently extracting maca polysaccharide.

Background

Maca (with the scientific name of Lepidium meyenii Walp, also called maca) is a cruciferous plant native to the mountain of Peru Andes, has oval leaves and roots similar to that of radish, is edible, is a pure natural food, has rich nutrient components and is reputed by Nanmei ginseng. The hypocotyl of maca may be gold or yellowish, red, purple, blue, black or green. China successfully introduced in 2003, nowadays, planting industries of a certain scale have been formed in Yunnan, Xinjiang and other places in China, and the Ministry of health approves maca as a new resource food in 2011, and related products develop and design functional foods, cosmetics and other aspects. The literature reports that macamides, glucosinolates and maca polysaccharides have various effects of resisting fatigue, resisting tumors, regulating immunity, regulating endocrine and the like. Maca as a new resource food has widely received attention of health care industries at home and abroad.

Polysaccharides are polymeric carbohydrate macromolecules composed of glycosidically bonded sugar chains and at least more than 10 monosaccharides. The polysaccharide is one of biological macromolecules, the research is more and more intensive at present, and a large number of literature reports exist, and many natural polysaccharides have various biological activity functions, such as antioxidant activity, blood sugar and blood fat reducing efficacy, cancer cell growth inhibition and the like. The research on the function of polysaccharide and the development of products are more and more, and the research becomes a leading hot spot of food science.

Disclosure of Invention

The aim of the part is to provide a method for efficiently extracting maca polysaccharide. The method uses maca as raw material, and separates three polysaccharide components with high purity, wherein the three polysaccharide components have good oxidation resistance and blood fat reducing effect.

In order to achieve the purpose, the invention comprises the following technical scheme:

a method for efficiently extracting maca polysaccharide comprises the following steps:

I. crushing and sieving maca roots and stems, and extracting water-soluble substances in maca powder by adopting an ultrasonic-assisted hot water extraction method;

II, filtering the mixture obtained in the step I to obtain a maca water extract, carrying out enzymolysis by sequentially adopting amylase and glucoamylase to remove starch, then inactivating the enzymes, and centrifuging to obtain a supernatant;

III, adding ethanol into the supernatant obtained in the step II, and standing at a low temperature to precipitate polysaccharide;

and IV, re-dissolving the precipitate obtained in the step III with water, removing protein from the re-dissolved solution by adopting a Sevage method, and freeze-drying to obtain the maca crude polysaccharide.

In the method, preferably, the specific operation of step I is: the extraction temperature is 40-60 ℃, the extraction time is 30-60min, the ultrasonic power is 150-300W, and the material-liquid ratio (weight/volume) is 1: 20-40 g/mL.

The method as described above, preferably, the specific operation of step II is: adding amylase into a maca water extract, wherein the mass ratio of the amylase to the raw maca powder is 1: 100-2000, and reacting for 2-5 hours at 30-80 ℃; and adding saccharifying enzyme, wherein the mass ratio of the saccharifying enzyme to the raw material maca powder is 1: 50-1000, reacting for 1-4 h at 30-80 ℃, then heating at 90-100 ℃ for 5-30 min to inactivate the enzyme, and centrifuging to obtain supernatant.

The method as described above, preferably, the specific operation of step II is: adding amylase into a maca water extract, wherein the mass ratio of the amylase to the raw maca powder is 1: 1000; reacting at 60 ℃ for 3h, adding saccharifying enzyme, reacting at 60 ℃ for 2h, heating at 100 ℃ for 10min to inactivate enzyme, and centrifuging to obtain supernatant.

The method as described above, preferably, the amylase of step II is an alpha amylase.

In the method, in the step III, the polysaccharide solution and the ethanol are preferably placed at 4-10 ℃ for 10-14 h at a volume ratio of 1: 2-4, so that the polysaccharide is precipitated and separated.

The method as described above, preferably, the specific operation of step IV is: preparing a Sevage reagent, wherein the volume ratio of n-butyl alcohol to chloroform is 1: 2-5; mixing and shaking the Sevage reagent and the polysaccharide solution according to the volume fraction of 1: 1-4, centrifuging and layering, reserving the upper layer solution, and then adding the Sevage reagent again for repeated operation until no protein is detected in the upper layer solution.

The method as described above, preferably, the method further comprises:

step V: and (3) separating the maca crude polysaccharide obtained in the step (IV) by using an anion exchange column to obtain three polysaccharide components, namely MPS1, MPS2 and MPS 3.

The method as described above, preferably, the anion exchange column separation conditions are:

the ion exchange column adopts polymethyl methacrylate resin as the filler and ddH as the solvent₂O, 0.1M NaCl and 0.2M NaCl; ddH₂O elution gave MPS1, 0.1M NaCl elution gave MPS2, and 0.2M NaCl elution gave MPS 3.

As in the above method, preferably, the MPS1 has a monosaccharide composition and mass ratio of arabinose, galactose, glucose, and mannose of 1.9: 1.4: 3.8: 1; the monosaccharide composition and the mass ratio of MPS2 are arabinose, galactose and glucose which are 1.8: 1.3: 1; the monosaccharide composition and mass ratio of MPS3 are rhamnose, arabinose, galactose and glucose 1: 15.6: 24.3: 5.3.

The invention has the beneficial effects that: the method has the advantages that the polysaccharide separated from the maca serving as the raw material mainly comprises three polysaccharide components, the method is simple to operate, the extraction temperature is low, the extraction time is short, the extraction rate is high, the purity is high, and the obtained three polysaccharide components have good oxidation resistance and blood fat reducing effects.

Drawings

FIG. 1 is a diagram showing the procedure of gradient elution of crude polysaccharide with an aqueous solution of sodium chloride using an anion exchange column.

FIG. 2 shows the results of absorbance measurements of three polysaccharide components.

FIG. 3 shows the result of DPPH radical scavenging experiment.

FIG. 4 shows the results of hydroxyl radical scavenging experiments.

Fig. 5 is an infrared spectrum of three maca polysaccharides.

Figure 6a is the monosaccharide composition of MPS 1;

figure 6b is the monosaccharide composition of MPS 2;

figure 6c is the monosaccharide composition of MPS 3.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof are described in detail below with reference to examples of the specification.

In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.