阅读说明：本技术 一种无患子皂苷水提取液中蛋白质的分离方法 (Method for separating protein in soapberry saponin water extract ) 是由 熊远钦 孙拥军 李官跃 于 2021-09-14 设计创作，主要内容包括：本发明提供了一种无患子皂苷水提取液中蛋白质的分离方法,属于化工分离工程领域,涉及日化原料中杂质分离技术问题。该分离方法包括以下步骤：⑴在无患子皂苷的水提取液中,加入葡萄糖酸-δ-内酯溶液、熟石膏溶液,充分搅拌、分散均匀；⑵加热处理液至50℃～60℃、保温,调慢搅拌速度,继续缓慢搅拌10～60分钟,⑶冷却至室温,同时静置150～180min后,用虹吸法抽取上部澄清液,再做离心沉降；二次沉降静置后虹吸的澄清液即为成品。⑷将各次虹吸上清液后剩余的“沉降渣”掏出,用板框压滤机压榨,此压滤液作为第⑴步水提取液复用。⑸压榨得到的滤饼,可作为有机肥料或土壤改良剂,提供给农资、园艺部门有效利用。(The invention provides a method for separating protein from a soapberry saponin water extracting solution, belongs to the field of chemical separation engineering, and relates to the technical problem of impurity separation in daily chemical raw materials. The separation method comprises the following steps: the preparation method comprises the steps of adding a gluconic acid-delta-lactone solution and a plaster of paris solution into a water extracting solution of sapindus saponin, fully stirring and uniformly dispersing; heating the treatment fluid to 50-60 ℃, preserving heat, slowing down the stirring speed, continuously and slowly stirring for 10-60 minutes, cooling to room temperature, standing for 150-180 minutes, extracting upper clear liquid by a siphoning method, and performing centrifugal sedimentation; standing for the second time, and siphoning to obtain the final product. And fourthly, taking out residual 'settled slag' after each siphoning of the supernatant, and squeezing the residual 'settled slag' by using a plate-and-frame filter press, wherein the pressure filtrate is reused as the water extracting solution in the first step. The filter cake obtained by the squeezing can be used as an organic fertilizer or a soil conditioner and is provided for effective utilization by agricultural and horticultural departments.)

1. A method for separating protein from a sapindus saponin water extract comprises the following steps of adding a gluconic acid-delta-lactone solution and a plaster of paris solution into the sapindus saponin water extract, and fully stirring and uniformly dispersing the mixture; heating the treatment fluid to 50-60 ℃, preserving heat, slowing down the stirring speed, continuously and slowly stirring for 10-60 minutes, cooling to room temperature, standing for 150-180 minutes, extracting upper clear liquid by a siphoning method, and performing centrifugal sedimentation; standing for the second time, and siphoning to obtain the final product. And fourthly, taking out residual 'settled slag' after each siphoning of the supernatant, and squeezing the residual 'settled slag' by using a plate-and-frame filter press, wherein the pressure filtrate is reused as the water extracting solution in the first step. The filter cake obtained by the squeezing can be used as an organic fertilizer or a soil conditioner and is provided for effective utilization by agricultural and horticultural departments.

2. The method for separating protein from sapindoside aqueous extract according to claim 1, wherein the glucono-delta-lactone solution is added in an amount of 0.5% to 10% in a concentration of 20% to 50% (based on the weight of sapindoside aqueous extract, the same applies hereinafter).

3. The method for separating protein from sapindus saponin aqueous extract according to claim 1, wherein the added aqueous solution of plaster of paris is 10% to 40% in an amount of 0.2% to 5%.

4. The method for separating protein from sapindus saponin aqueous extract according to claim 1, wherein the coagulant and the coagulant aid are added, the temperature of the heat-preservation treatment solution is 50 ℃ to 60 ℃, and the mixture is slowly stirred for 10 to 60 minutes, so that the protein in the sapindus saponin aqueous extract is in full contact with the gluconic acid-delta-lactone and the plaster of paris to form a flocculated form and aggregate.

5. The method for separating protein from sapindoside water extract according to claim 1, wherein after flocculation and aggregation of protein, the treated solution is cooled to room temperature, left to stand for 150-180 min, and then upper clear solution is siphoned off and settled (centrifuged); and (4) siphoning the supernatant liquid after secondary sedimentation to obtain a finished product.

Technical Field

The invention provides a method for separating protein in soapberry saponin water extract. Belongs to the field of biological resource utilization and light chemical engineering application, and relates to the technical problems of chemical engineering separation engineering and protein separation.

Technical Field

Sapindus mukorossi is also known as "hand washing fruit", taro, soapberry. The wood stick made of the disease-free tree can repel the evil and kill the ghost, so the disease-free wood stick is named as disease-free. The plant is cultivated in Fujian, Guangdong, Guangxi, Jiangxi, Zhejiang and other areas in China, has the advantages of fast growth, strong wind resistance, low requirement on soil, water and soil conservation function and strong resistance to sulfur dioxide, and is frequently used as an urban greening tree species. The soapberry saponin extract takes triterpenoid structural saponin as a main component, is from nature, is easy to degrade by natural microorganisms, and has the advantages of effective utilization of resources and short cycle regression period. As a natural surfactant, the surfactant has wide application in a plurality of fields such as daily chemical industry, biological pesticide, tertiary oil recovery, industrial cleaning and the like.

The extraction process of the soapberry saponin mainly takes ethanol, water, methanol and butanol as solvents for extraction at present. Due to the difference in solubility, the alcohol-extracted product has lower protein and carbohydrate impurities and lighter color than the water-extracted product; but the yield of the total saponins is low, the production cost is high, and the danger and environmental protection problems in the production process are more prominent. Therefore, the trend in recent years is toward aqueous extraction.

The molecular structures of the sapindoside, the protein and the carbohydrate all contain polar hydrophilic groups, and the solubility in water is very close, so that the contents of the three substances in the sapindoside solution stock solution extracted by the water method are basically close (about 7%, 6% and 8% in sequence). Although the coexistence introduction of protein and saccharide is tolerable in many sapindoside applications, the method is disadvantageous in terms of product storage, formula design, antisepsis, bacteria prevention and pollution prevention, and the content of protein and saccharide in sapindoside water extract needs to be reduced as much as possible.

Glucono-delta-lactone, lactone or GDL for short, molecular formula C₆H₁₀O₆Relative molecular mass 178.14. White crystal or white crystalline powder, almost odorless, sweet first and sour second. Easily dissolved in water (60g/100 mL). Glucono-delta-lactone can be used as a protein coagulant. The toxicological test data are as follows: LD50 rabbit intravenous injection 7.63g/kg body weight, GRAS FDA-21CFR 184.1318, ADI do not do requirements (FAO/WHO, 1994), adult volunteers take the product (dosage is 167mg/kg body weight), 7.7% -15% of metabolism is excreted by urine after 7h, and no abnormality of urine is found. As a food additive, the maximum use amount of the food additive is 0.1g/kg specified in the food additive use sanitary Standard of China (GB2760-2011), and the food additive is used for keeping fish and shrimp fresh and is used for sausages (sausages), meat paste products, grape juice and bean products (bean curd and beans)Flowers) at a maximum of 3.0 g/kg.

Calcined gypsum Ca [ SO ]₄]·0.5H₂O, also known as plaster of paris or hemihydrate. The food additive is listed as a food additive sequence by using the sanitary standard in China, can be used as a protein coagulant aid in the production of bean products, and can be added according to the normal production requirement.

Disclosure of Invention

The invention discloses a method for separating protein in soapberry saponin water extract, which is based on the flocculation of gluconic acid-delta-lactone on protein, is assisted by plaster of paris as a coagulant aid, is mixed and dispersed, and is fully contacted with protein molecules in the soapberry saponin water extract, so that the protein is aggregated in a flocculation form, and is separated by sedimentation (or centrifugation). It includes but is not limited to the following process steps: 5 to 100 parts of gluconic acid-delta-lactone aqueous solution with the concentration of 20 to 50 percent and 2 to 50 parts of plaster of paris aqueous solution with the concentration of 10 to 40 percent are added into 1000 parts of soapberry saponin aqueous extract which is primarily settled (or centrifugally) separated (parts by weight, the same below), and the materials are added and stirred. After the coagulant and the coagulant aid are added, the treated solution is heated to 50-60 ℃, the temperature is kept, the stirring speed is slowed down, and the slow stirring is continued for 10-60 minutes, so that the protein in the soapberry saponin water extract, the gluconic acid-delta-lactone and the plaster of paris achieve full contact of molecular level, and the flocculation form is formed and is aggregated. The slow agitation favours the coagulation and aggregation of the proteins and "frees" the sapindoside, which may be entrapped in the flocculation, back into the extraction solution. Cooling the treated liquid to room temperature, standing for 150-180 min, extracting upper clear liquid by a siphon method, and performing (centrifugal) sedimentation; and (4) siphoning the supernatant liquid after secondary sedimentation to obtain a finished product. Taking out the residual settling dregs after siphoning the supernatant, squeezing with a plate-and-frame filter press, and recovering the filtrate as soapberry saponin water extract for reuse. The filter cake obtained by squeezing can be used as organic fertilizer or soil conditioner, and can be effectively utilized by gardening and agricultural material departments.

The invention has the following beneficial effects:

1. the gluconic acid-delta-lactone and the plaster belongs to food additives, are non-toxic and harmless, and can enhance the coagulation effect of protein and improve the separation efficiency of the protein under the synergistic action.

2. After the coagulant and the coagulant aid are added, the treatment liquid is heated and slowly stirred, so that the coagulation and aggregation of the protein can be promoted, the soapnut saponin possibly wrapped in the flocculation group is enabled to be dissociated and return to the extracting solution, and the yield of the soapnut saponin is improved.

3. The later standing and siphon extraction of the treatment liquid can effectively slow down the blockage of floc to the filter cloth in the subsequent filter pressing process, and reduce the filtering workload and the entrainment loss of sapindus saponin in the filter cake.

Description of the drawings:

FIG. 1 shows a comparison of technical data of "example" and "comparative example" in this patent.

The specific implementation mode is as follows:

the technical solution of the present invention is further illustrated by the following examples.

Example 1

10kg of gluconic acid-delta-lactone aqueous solution with the concentration of 38 percent and calcined gypsum aqueous solution with the concentration of 25 percent are respectively dissolved and prepared in two batching barrels in advance for standby. Then, 800kg of the preliminarily settled (or centrifuged) soapberry saponin aqueous extract is put into a mixing kettle with the volume of 1 ton, the stirring is started, and 10kg of the gluconic acid-delta-lactone aqueous solution and 3kg of the calcined gypsum aqueous solution are added under the stirring. After the coagulant and the coagulant aid are added, the treatment solution is heated to 50 ℃ to 60 ℃, then the stirring speed is reduced, the stirring is slowly carried out for 40 minutes, and the protein in the soapberry saponin water extract, the gluconic acid-delta-lactone and the plaster of paris are in full contact at a molecular level to form a flocculation shape and aggregate. Further cooling the treated liquid to room temperature, standing for 180min, extracting upper clear liquid by a siphon method, transferring the upper clear liquid into another material tank, and performing sedimentation separation; and (4) after secondary sedimentation, siphoning to obtain a clear liquid, namely the finished product of the soapberry saponin water extract. Through detection, the finished product liquid has the protein content of 0.58 percent (the protein content of the extracting solution before separation treatment is 6.7 percent), has clear and stable color, and meets the quality requirement of the national forestry industry standard LY/T3105- "Sapindus saponin" of the people's republic of China.

Example 2

Similarly to example 1, 38% aqueous glucono-delta-lactone solution and 25% aqueous plaster solution were dissolved and prepared in two preparation tanks, respectively, and were used. 1000 parts (by weight, the same below) of soapberry saponin aqueous extract of protein to be separated, 18 parts of the gluconic acid-delta-lactone aqueous solution and 5.5 parts of calcined gypsum aqueous solution are sequentially added into a mixing kettle according to the following proportion, and the materials are added while stirring. After the materials are added, the treatment solution is heated to 50-60 ℃, the temperature is kept, the stirring speed is slowed down, and the slow stirring is continued for 60 minutes, so that the protein in the soapberry saponin water extract forms a flocculation shape and is aggregated. Then cooling the treated liquid to room temperature, standing for 150min, and extracting supernatant liquid from the upper part by a siphon method and introducing the supernatant liquid into a centrifugal machine for centrifugal sedimentation; and (4) siphoning the supernatant liquid after secondary sedimentation to obtain the soapberry saponin solution product. Taking out the residual settling dregs after siphoning the supernatant, squeezing with a plate-and-frame filter press, and recovering the filtrate as soapberry saponin water extract for reuse. The filter cake obtained by squeezing can be used as organic fertilizer or soil conditioner, and can be effectively utilized by gardening and agricultural material departments.

Through detection, the finished product liquid has the protein content of 0.76 percent (the protein content of the extracting solution before separation treatment is 6.7 percent), has clear and stable color, and meets the technical requirements of the industry standard LY/T3105- "Sapindus saponin" of forestry industry of the people's republic of China.

Comparative example 1

This comparative example is exemplified by the Chinese patent application No. 201510126843.0 entitled Sapindus saponin solution containing a clear and stable compound and its preparation method. The method comprises the following steps: (1) adding a compound clarifying agent into an enzymolysis saponin extracting solution of the soapberry peel, and standing for 30-60 min; (2) adding a compound antioxidant into the saponin extracting solution, heating to 90-100 ℃, and preserving heat for 30-40 min; (3) cooling the saponin extracting solution to room temperature, sealing, standing for 2-4 h, and obtaining a supernatant by a siphoning method; (4) making the supernatant fluid pass through a diatomite filter to prepare a primary clarified liquid; (5) and centrifuging the primary clarified liquid, and adding a clarified and stabilized compound to prepare a clarified and stabilized sapindus saponin liquid. The composite clarifying agent in the step (1) comprises the following components: 50-60 wt% of tannin, 10-20 wt% of seaweed gel and 20-30 wt% of gelatin; the addition amount of the composite clarifying agent is 5-8% of the weight of the saponin extracting solution. The composite antioxidant in the step (2) comprises the following components: 50-55 wt% of sodium erythorbate, 35-40 wt% of beta-carotene, 2-4 wt% of phytic acid and 3-6 wt% of polyvinylpyrrolidone; the adding amount of the compound antioxidant is 3-5% of the weight of the saponin extracting solution. The kieselguhr filter in the step (4) is characterized in that the upper end of the suction filtration container is connected with a Buchner funnel, filter paper is paved in the Buchner funnel, and kieselguhr with the thickness of 1-2 cm is paved on the filter paper; washing the diatomite with deionized water, and filtering to obtain diatomite filtering foam above the filter paper. The clear stable compound in the step (5) comprises the following components: 10-15 wt% of sodium erythorbate, 10-15 wt% of isomeric tridecanol polyoxyethylene ether, 105-8 wt% of polyglycerol, 40-50 wt% of dipropylene glycol, 18-25 wt% of propylene glycol and 0.001-0.005 wt% of methylisothiazolinone; the addition amount of the flocculant is 5-10% of the weight of the primary clarified liquid.

For comparison, the applicant takes the "median" of the composition range of each raw material in the above-mentioned chinese patent 201510126843.0 as the ratio, and processes the sapindoside water extract of the protein to be separated according to the preparation procedure to obtain a clear and stable sapindoside solution. The detection is carried out according to the forestry industry standard LY/T3105-.

TABLE 1 comparison of technical data of "examples" and "comparative examples" of this patent

Comparing the test technical data of the 2 embodiments and the comparative example, it can be easily seen that by adopting the technical scheme of the invention, the produced products all meet the regulations of the existing forestry industry standard LY/T3105- "Sapindus saponin" of the people's republic of China; compared with the imitation process product (namely a comparative example) of Chinese invention patent 201510126843.0, a sapindus saponin liquid containing a clear and stable compound and a preparation method thereof, the imitation process product can meet the technical requirements of the industrial standard on the indexes of total saponin content, critical micelle concentration, water insoluble content, pH value of 1% water solution and solid content, and has slight advantages in data (belonging to the measurement range of the same level). Compared with the comparative example, the patent example has the obvious advantages that the appearance of the obtained treatment fluid is light in color, the turbidity phenomenon is not easy to occur, the solid content and the protein content are lower, and the foaming power is higher.

Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.