阅读说明：本技术 一种提取姜黄甾体皂苷的方法 (Method for extracting steroid saponin from curcuma longa ) 是由 朱继强 于 2021-06-02 设计创作，主要内容包括：本发明公开了一种提取姜黄甾体皂苷的方法,由以下步骤制备：S1,通过清洗打浆器清洗,打浆,得黄姜浆液；S2,通过混合物短时发酵,并通过十二烷基磺酸钠,增大与水中的混合能力；S3,然后加入黑曲霉菌悬液,二次发酵；S4,加入过量铁粉,生成含有葡萄糖酸亚铁,除去十二烷基磺酸钠,然后通过超疏油滤膜分离,得到姜黄甾体皂苷粗体物和葡萄糖酸亚铁溶液,分别精制,得姜黄甾体皂苷和葡萄糖酸亚铁。该方法所提取的姜黄甾体皂苷的产量高,无酸及有机溶剂的添加,避免了对环境的污染,真正实现了绿色提取,同时将淀粉转化利用生成葡萄糖酸亚铁,实现了对淀粉的高效利用。(The invention discloses a method for extracting steroid saponin of turmeric, which comprises the following steps: s1, washing and pulping by a washing and pulping device to obtain yellow ginger pulp; s2, the mixture is fermented for a short time, and the mixing capacity of the mixture and water is increased through sodium dodecyl sulfate; s3, adding aspergillus niger suspension, and performing secondary fermentation; s4, adding excessive iron powder to generate ferrous gluconate, removing sodium dodecyl sulfate, separating with superoleophobic filter membrane to obtain Curcuma rhizome steroid saponin coarse product and ferrous gluconate solution, and refining to obtain Curcuma rhizome steroid saponin and ferrous gluconate. The method has the advantages of high yield of the extracted steroid saponins of Curcuma longa, no addition of acid and organic solvent, no environmental pollution, real realization of green extraction, and realization of high-efficiency utilization of starch by converting and utilizing starch to generate ferrous gluconate.)

1. The method for extracting the steroid saponins of turmeric is characterized by comprising the following steps:

s1, inserting and fixing the stem of a fresh turmeric rhizome into an insertion tube of a cleaning and pulping device, starting a motor, enabling the turmeric rhizome to do self circular motion, starting a spray cleaning device, cleaning the turmeric rhizome, sterilizing at the high temperature of 80-100 ℃ for 2-3min, starting a telescopic rod, driving a circular blade to cut the turmeric rhizome from the stem, enabling the turmeric rhizome to fall into the bottom of the cleaning and pulping device, closing a stop valve, starting a chopping and pulping device, and pulping to obtain turmeric pulp;

s2, adding the Bacillus belgii bacterial suspension, the Bacillus subtilis bacterial suspension and the white rot fungi bacterial suspension into the turmeric slurry obtained in the step S1, performing ventilation flow fermentation at 30-35 ℃ for 2-3d, then adding sodium dodecyl sulfate into the fermentation liquor, increasing the dissolving capacity of the turmeric steroid saponin in water, stirring for 10-20min, then filtering the fermentation liquor, washing the filter residue with deionized water (50 mL multiplied by 3), and combining the fermentation filtrate and the washing liquor to obtain a mixed liquor 1, wherein the main components of the mixed liquor 1 comprise the turmeric steroid saponin, glucose, sodium dodecyl sulfate, Bacillus belgii, the Bacillus subtilis, the white rot fungi and the deionized water;

s3, adding aspergillus niger suspension into the mixed solution 1, carrying out sealed fermentation for 4-5 days at 28-32 ℃, adding primary deionized water every 5 times in the fermentation process to obtain secondary fermentation liquor, sterilizing the secondary fermentation liquor at 80-100 ℃, and filtering thalli through an ultrafiltration membrane to obtain a mixed solution 2, wherein the main components of the mixed solution 2 comprise spiro-type curcuma longa steroid saponin, gluconic acid, sodium dodecyl sulfate and deionized water;

s4, adding excessive iron powder into the mixed solution 2, heating at 50-60 ℃, stirring at 2000-2500rpm to generate a reaction solution containing ferrous gluconate, sodium dodecyl sulfate and Fe²⁺The complex forms foam floating on the surface of the reaction liquid due to violent stirring, the foam on the surface of the reaction liquid is scraped, the excessive iron powder is removed by filtration, and then the mixture is separated by a super-oleophobic filter membrane to obtain turmeric steroid saponin coarse substances and ferrous gluconate solution which are respectively refined to obtain turmeric steroid saponin and ferrous gluconate.

2. The method for extracting curcuminoid saponin according to claim 1, wherein the mass volume ratio of fresh turmeric rhizome to deionized water in turmeric slurry is 2-2.2kg: 2-2.2L.

3. The method for extracting steroidal saponins of curcuma longa according to claim 1, wherein the mass-to-volume ratio of bacillus beijerinckii suspension, bacillus subtilis suspension, white rot fungus suspension and turmeric slurry in step S2 is 10-12mL, 18-20mL, 2.2-2.4L, and the total effective viable count of bacillus beidelukii suspension is not less than 1.0 x 10⁶cfu/mL, the total effective viable count of the bacillus subtilis suspension is not less than 1.5 multiplied by 10⁶cfu/mL, the total effective viable count of the white rot fungus suspension is not less than 1.4 multiplied by 10⁶cfu/mL; the mass volume ratio of the fermentation liquor to the sodium dodecyl sulfate is as follows: 2.2-2.4L:350-400g。

4. The method for extracting steroidal saponins of curcuma longa according to claim 1, wherein the mass-to-volume ratio of mixed solution 1 to aspergillus niger suspension to deionized water in step S3 is: 20-25mL of 2.3L-2.5L and 50-80mL of the total weight of the mixture; the total effective viable count of Aspergillus niger suspension is not less than 1.8 × 10⁶ cfu/mL。

5. The method for extracting curcuminoid saponin according to claim 1, wherein the crude curcuminoid saponin is recrystallized, dried and refined to obtain curcuminoid saponin solid.

6. The method for extracting curcuminoid steroid saponin according to claim 1, wherein the ferrous gluconate solution is refined by decoloring, crystallizing and drying to obtain ferrous gluconate solid.

7. The method for extracting steroid saponins from curcuma longa according to claim 1, wherein the cleaning beater comprises a housing, a circular temperature-controlled heating shell is sleeved on the housing, the circular temperature-controlled heating shell and the housing can be opened/sealed along any diameter, a telescopic cylinder is arranged on the top of the housing, the telescopic cylinder is connected with a circular blade, the telescopic cylinder drives the circular blade to perform vertical cutting movement, a plurality of spraying and washing devices are uniformly distributed in the middle of the housing, an outlet pipeline and a crushing and beating device are arranged at the bottom of the housing, a stop valve is arranged on the outlet pipeline, a cylindrical triangular bracket is arranged at the middle lower part of the housing, a motor is arranged on the triangular bracket, a first bevel gear is connected on the motor, a plurality of second bevel gears are engaged on the first bevel gear, an insertion tube is arranged on any second bevel gear, the motor drives the insertion tube to perform self-circular movement through the first bevel gear and the second bevel gear, the first bevel gear and the second bevel gear are provided with an upper cover plate and a lower cover plate, and the radius of the circular blade is larger than and close to the sum of the length of the insertion pipe and the radius of the upper cover plate.

Technical Field

The invention relates to the field of extraction of steroid saponins, in particular to a method for extracting steroid saponins from turmeric.

Background

Yellow ginger, known as dioscorea zingiberensis, is a unique medicinal plant resource in China. The yellow ginger is mainly used for producing yellow ginger saponin, and the yellow ginger saponin can be used for synthesizing a plurality of steroid hormone medicaments and is known as medicinal gold. The turmeric saponin exists in the rhizome of the turmeric in the form of turmeric steroid saponin, the turmeric steroid saponin is an amphiphilic compound with glycosyl groups connected at C3 and C26 of the turmeric saponin, all components in the turmeric aqueous suspension are generally acidolyzed together in industry to produce the turmeric steroid saponin, bottlenecks of large pollution, low yield, high cost and the like exist, in addition, the rhizome of the turmeric also contains a large amount of starch and lignocellulose, and the turmeric rhizome is hydrolyzed by acid to become pollutants in production wastewater, so that the treatment difficulty of the production wastewater is increased, and the serious waste of starch and lignocellulose resources is caused.

At present, China develops a great deal of work around the research and development of the clean production technology of diosgenin, and also makes some progress, and forms the following improved production processes: (1) a direct separation method; (2) a saccharification membrane separation method; (3) alcohol extract method. Although the improved process reduces the use of acid and achieves some results, the acid hydrolysis is still needed, and the green production of the diosgenin cannot be realized in a real sense.

Disclosure of Invention

In view of the above circumstances, an object of the present invention is to provide a method for extracting turmeric steroid saponins from fresh turmeric rhizome, which can extract turmeric steroid saponins from fresh turmeric rhizome in a green, efficient and sufficient manner, and can effectively utilize starch and lignocellulose resources therein.

The technical proposal for solving the problem is that,

a method for extracting steroid saponin from Curcuma rhizome comprises the following steps:

s1, inserting and fixing the stem of a fresh turmeric rhizome into an insertion tube of a cleaning and pulping device, starting a motor, enabling the turmeric rhizome to do self circular motion, starting a spray cleaning device, cleaning the turmeric rhizome, sterilizing at the high temperature of 80-100 ℃ for 2-3min, starting a telescopic rod, driving a circular blade to cut the turmeric rhizome from the stem, enabling the turmeric rhizome to fall into the bottom of the cleaning and pulping device, closing a stop valve, starting a chopping and pulping device, and pulping to obtain turmeric pulp;

s2, adding the Bacillus belgii bacterial suspension, the Bacillus subtilis bacterial suspension and the white rot fungi bacterial suspension into the turmeric slurry obtained in the step S1, performing ventilation flow fermentation at 30-35 ℃ for 2-3d, then adding sodium dodecyl sulfate into the fermentation liquor, increasing the dissolving capacity of the turmeric steroid saponin in water, stirring for 10-20min, then filtering the fermentation liquor, washing the filter residue with deionized water (50 mL multiplied by 3), and combining the fermentation filtrate and the washing liquor to obtain a mixed liquor 1, wherein the main components of the mixed liquor 1 comprise the turmeric steroid saponin, glucose, sodium dodecyl sulfate, Bacillus belgii, the Bacillus subtilis, the white rot fungi and the deionized water;

s3, adding aspergillus niger suspension into the mixed solution 1, carrying out sealed fermentation for 4-5 days at 28-32 ℃, adding primary deionized water every 5 times in the fermentation process to obtain secondary fermentation liquor, sterilizing the secondary fermentation liquor at 80-100 ℃, and filtering thalli through an ultrafiltration membrane to obtain a mixed solution 2, wherein the main components of the mixed solution 2 comprise spiro-type curcuma longa steroid saponin, gluconic acid, sodium dodecyl sulfate and deionized water;

s4, adding excessive iron powder into the mixed solution 2, heating at 50-60 ℃, stirring at 2000-2500rpm to generate a reaction solution containing ferrous gluconate, sodium dodecyl sulfate and Fe²⁺The complex forms foam floating on the surface of the reaction liquid due to violent stirring, the foam on the surface of the reaction liquid is scraped, the excessive iron powder is removed by filtration, and then the mixture is separated by a super-oleophobic filter membrane to obtain turmeric steroid saponin coarse substances and ferrous gluconate solution which are respectively refined to obtain turmeric steroid saponin and ferrous gluconate.

Furthermore, the mass volume ratio of the fresh turmeric rhizome to the deionized water in the turmeric slurry is 2-2.2kg: 2-2.2L.

Further, in the step S2, the mass-volume ratio of the Bacillus belgii bacterial suspension to the Bacillus subtilis bacterial suspension to the white rot fungi bacterial suspension to the turmeric slurry is 10-12mL to 18-20mL to 2.2-2.4L, and the total effective viable count of the Bacillus belgii bacterial suspension is not less than 1.0 multiplied by 10⁶cfu/mL, total effective activity of Bacillus subtilis suspensionThe number of bacteria is not less than 1.5 × 10⁶cfu/mL, the total effective viable count of the white rot fungus suspension is not less than 1.4 multiplied by 10⁶cfu/mL; the mass volume ratio of the fermentation liquor to the sodium dodecyl sulfate is as follows: 2.2-2.4L:350-400 g.

Further, in the step S3, the mass-to-volume ratio of the mixed solution 1 to the aspergillus niger suspension to the deionized water is as follows: 20-25mL of 2.3L-2.5L and 50-80mL of the total weight of the mixture; the total effective viable count of Aspergillus niger suspension is not less than 1.8 × 10⁶ cfu/mL。

Further, the crude curcuma steroid saponin is recrystallized, dried and refined to obtain solid curcuma steroid saponin.

Further, the ferrous gluconate solution is decolored, crystallized, dried and refined to obtain ferrous gluconate solid.

Furthermore, the cleaning beater comprises a casing, a circular temperature-control heating casing positioned outside the casing is sleeved on the casing, the circular temperature-control heating casing and the casing can be opened/sealed along any diameter, a telescopic cylinder is arranged at the top of the casing and connected with a circular blade, the telescopic cylinder drives the circular blade to do vertical cutting motion, a plurality of uniformly-distributed spray washing devices are arranged in the middle of the casing, an outlet pipeline and a crushing beater are arranged at the bottom of the casing, a stop valve is arranged on the outlet pipeline, a cylindrical triangular support is arranged at the middle lower part of the casing, a motor is arranged on the triangular support, a first bevel gear is connected onto the motor, a plurality of second bevel gears are meshed onto the first bevel gear, an insert pipe is arranged on any second bevel gear, the motor drives the insert pipe to do self circular motion through the first bevel gear and the second bevel gears, and an upper cover plate and a lower cover plate are arranged on the first bevel gear and the second bevel gears, the radius of the circular blade is larger than and close to the sum of the length of the insertion tube and the radius of the upper cover plate.

According to the invention, the turmeric steroid saponin is fully dispersed in a water phase, and then a target product is collected through a water-oil separation membrane, and meanwhile, a byproduct is collected and utilized.

The Curcuma rhizome steroid saponin includes furostan type saponin and spirostan type saponin. The furostane saponin is water soluble saponin, most of the spirostane saponin is insoluble in water, free furostane saponin and water soluble spirostane saponin are all present in the water phase of the water suspension, and unreleased furostane saponin and all water insoluble spirostane saponin are present in the solid phase of the water suspension. Therefore, part of the saponins in the rhizome of turmeric cannot be dissolved or dispersed in the water phase, so that a large amount of saponins exists in the solid phase, and a large amount of organic solvents are still needed for extraction in the subsequent step, so that the production cost is high, and the implementation of the process of efficiently converting the saponins into the saponins by the microorganisms in the next step is not facilitated. Therefore, how to ensure the stability of the content of the water-soluble saponins in the yellow ginger and how to efficiently promote the saponin in the solid phase of the yellow ginger to be released into the water phase are one of the key problems for improving the yield of the steroid saponins of the yellow ginger.

The interaction of endogenous enzyme and surface microorganism of the rhizome of yellow ginger can affect the stability of saponin in the rhizome of yellow ginger, and convert furostane type saponin into spirostane type saponin, so when soil impurities on the surface of the rhizome of yellow ginger are fully removed by the cleaning beater, the epidermis integrity of the rhizome of yellow ginger can be protected as much as possible, the invasion of the surface microorganism is prevented, meanwhile, the activity of endogenous enzyme and surface microorganism of the rhizome of yellow ginger can be further reduced by high-temperature sterilization, the cleaning, crushing and pulping operations are finished in the same space, the opportunity of external microorganism entering is further reduced, and the stability of saponin in the rhizome of yellow ginger is maintained to the maximum extent.

The turmeric rhizome is mainly composed of periderm, basic tissue and scattered vascular bundles, turmeric steroid saponin is mainly present in parenchyma cells of the basic tissue, living cells of phloem and vascular bundle sheath cells also have accumulation of the turmeric steroid saponin, the saponin is present in specific tissue cells of plants and is wrapped by lignocellulose and starch components in the turmeric rhizome, therefore, in order to fully release the steroid saponin in the turmeric rhizome, the invention adopts Bacillus belgii, Bacillus subtilis and white rot fungi to ferment turmeric pulp for a short time, the Bacillus belgii and the white rot fungi degrade lignin cellulose in the turmeric rhizome to form a loose structure, and the Bacillus subtilis decomposes the starch in the turmeric rhizome into glucose.

The saponin in the fresh yellow ginger rhizome mainly comprises furostane-type saponin and spirostane-type saponin, so that the saponin in the water phase comprises all water-soluble saponin and a part of saponin which can form saponin microcapsules, namely the furostane-type saponin and a part of spirostane-type saponin, and a part of the yellow ginger spirostane-type saponin is not in the water phase.

According to the method, glucose in the mixed solution 1 is converted into gluconic acid through secondary fermentation of aspergillus niger, furostane-type saponin can be converted into spirostane-type saponin through acidity of the solution, water solubility of turmeric steroid saponin is reduced, ferrous gluconate is obtained through reduction of gluconic acid and iron powder, generated ferrous ions are complexed with sodium dodecyl sulfate, and foam is formed to scrape off the complex, so that dispersity of the turmeric steroid saponin and water is further reduced, an oil phase and a water phase are separated through a super-oleophobic filter membrane, loss of oil-water separation operation is reduced, high-yield spiro-type turmeric steroid saponin is obtained finally, and starch byproducts are effectively utilized to convert to obtain the ferrous gluconate.

Due to the adoption of the technical scheme, compared with the prior art, the invention has the following advantages;

1. the turmeric steroid saponin extracted by the method for extracting the turmeric steroid saponin has high yield.

2. According to the method for extracting the steroid saponins of turmeric, the addition of acid and the addition of an organic solvent in the traditional production are avoided, the pollution to the environment is avoided, and the green extraction is really realized.

3. The method for extracting the steroid saponins of turmeric not only realizes the high-yield extraction of the steroid saponins of turmeric, but also converts and utilizes starch to generate ferrous gluconate, thereby realizing the high-efficiency utilization of the starch.

Drawings

FIG. 1 is a view of the cleaning beater of the present invention;

FIG. 2 is a cross-sectional view of the cleaning beater of the present invention;

FIG. 3 is a view of the cleaning beater of the present invention;

fig. 4 is a preparation diagram of the invention for extracting the steroid saponin of turmeric.

Detailed Description

The foregoing and other technical and scientific aspects, features and utilities of the present invention will be apparent from the following detailed description of the embodiments, which is to be read in connection with the accompanying drawings of fig. 1-4. The structural contents mentioned in the following embodiments are all referred to the attached drawings of the specification.

Example 1

A method for extracting steroid saponin from Curcuma rhizome comprises the following steps:

s1, inserting 2kg of stems 11 of fresh turmeric rhizomes into an insertion tube 126 fixed on a cleaning beater, starting a motor 125, enabling the turmeric rhizomes to do self circular motion, starting a spray washing device 6, cleaning the turmeric rhizomes, sterilizing at 80 ℃ for 2min, starting a telescopic rod 3, driving a circular blade 5 to cut the turmeric rhizomes from the stems, enabling the turmeric rhizomes to fall into the bottom of the cleaning beater, closing a stop valve 9, spraying 2L of deionized water, starting a chopping beater 7, and beating to obtain turmeric slurry;

s2, adding 10mL of Bacillus belgii suspension, 10mL of Bacillus subtilis suspension and 18mL of white rot fungus suspension into the turmeric slurry obtained in the step S1, performing ventilation flow fermentation at 30 ℃ for 2d, adding 350g of sodium dodecyl sulfate into the fermentation liquor, increasing the dissolving capacity of turmeric steroid saponin in water, stirring for 10min, filtering the fermentation liquor, washing the filter residue with deionized water (50 mL multiplied by 3), and combining the fermentation filtrate and the washing liquor to obtain a mixed liquor 1, wherein the main components of the mixed liquor 1 comprise turmeric steroid saponin, glucose, sodium dodecyl sulfate, Bacillus belgii, Bacillus subtilis, white rot fungus and deionized water;

s3, adding 20mL of aspergillus niger suspension into 12.3 LL of the mixed solution, carrying out sealed fermentation for 4d at 28 ℃, adding 50mL of primary deionized water every 5 times in the fermentation process to obtain secondary fermentation liquor, sterilizing the secondary fermentation liquor at 80 ℃, and filtering thalli through an ultrafiltration membrane to obtain a mixed solution 2, wherein the main components of the mixed solution 2 comprise spiro-type curcuma longa steroid saponin, gluconic acid, sodium dodecyl sulfate and deionized water;

s4, adding excessive iron powder into the mixed solution 2, heating at 50 ℃, stirring at 2000rpm to generate a reaction solution containing ferrous gluconate, sodium dodecyl sulfate and Fe²⁺The complex forms foam floating on the surface of the reaction liquid due to violent stirring, the foam on the surface of the reaction liquid is scraped, the excessive iron powder is removed by filtration, then the mixture is separated by a superoleophobic filter membrane to obtain turmeric steroid saponin coarse substance and ferrous gluconate solution, the turmeric steroid saponin coarse substance and the ferrous gluconate solution are respectively refined to obtain turmeric steroid saponin and ferrous gluconate, and the turmeric steroid saponin and the ferrous gluconate solution are weighed.

The total effective viable count of the Bacillus beleisi bacterial suspension is not less than 1.0 x 10⁶cfu/mL, the total effective viable count of the bacillus subtilis suspension is not less than 1.5 multiplied by 10⁶cfu/mL, the total effective viable count of the white rot fungus suspension is not less than 1.4 multiplied by 10⁶cfu/mL; the total effective viable count of the aspergillus niger suspension bacterial suspension is not less than 1.8 multiplied by 10⁶ cfu/mL。

The crude curcuma longa steroid saponin is recrystallized, dried and refined to obtain solid curcuma longa steroid saponin; and the ferrous gluconate solution is subjected to decoloration, crystallization, drying and refining to obtain a ferrous gluconate solid.

The cleaning beater comprises a casing 2, a circular temperature-controlled heating shell 1 positioned outside the casing 2 is sleeved on the casing 2, the circular temperature-controlled heating shell 1 and the casing 2 can be opened/sealed along any diameter 4, a telescopic cylinder 3 is arranged at the top of the casing 2, the telescopic cylinder 3 is connected with a circular blade 5, the telescopic cylinder 3 drives the circular blade 3 to do up-and-down cutting motion, a plurality of uniformly-distributed spray washing devices 6 are arranged at the middle part of the casing 2, an outlet pipeline 8 and a crushing beater 7 are arranged at the bottom of the casing 2, a stop valve 9 is arranged on the outlet pipeline, a cylindrical triangular bracket 10 is arranged at the middle lower part of the casing 2, a motor 125 is arranged on the triangular bracket 10, a first bevel gear 124 is connected on the motor 125, a plurality of second bevel gears 123 are engaged on the first bevel gear 124, an insert pipe 126 is arranged on any second bevel gear 123, the motor 125 drives the insert pipe 126 to do self circular motion through the first bevel gear 124 and the second bevel gear 123, the first bevel gear 124 and the second bevel gear 123 are provided with an upper cover plate 121 and a lower cover plate 122, the upper cover plate 121 and the lower cover plate 122 just cover the first bevel gear 124 and the second bevel gear 123, and the radius of the circular blade 5 is larger than and close to the sum of the length of the insertion pipe 126 and the radius of the upper cover plate 121.

Example 2

A method for extracting steroid saponin from Curcuma rhizome comprises the following steps:

s1, inserting 2.1kg of stems 11 of fresh turmeric rhizome into an insertion tube 126 fixed on a cleaning beater, starting a motor 125, enabling the turmeric rhizome to do self circular motion, starting a spray washing device 6, cleaning the turmeric rhizome, then sterilizing at high temperature of 90 ℃ for 2.5min, starting a telescopic bar 3, driving a circular blade 5 to cut the turmeric rhizome from the stems, enabling the turmeric rhizome to fall into the bottom of the cleaning beater, closing a stop valve 9, spraying 2.1L of deionized water, starting a chopping beater 7, and beating to obtain turmeric pulp;

s2, adding 11mL of Bacillus belgii suspension, 11mL of Bacillus subtilis suspension and 19mL of white rot fungi suspension into the turmeric slurry obtained in the step S1, performing ventilation flow fermentation at 33 ℃ for 2.5 days, adding 375g of sodium dodecyl sulfate into the fermentation liquor, increasing the dissolving capacity of turmeric steroid saponin in water, stirring for 15min, filtering the fermentation liquor, washing filter residues with deionized water (50 mL multiplied by 3), and combining the fermentation filtrate and the washing liquor to obtain a mixed liquor 1, wherein the main components of the mixed liquor 1 comprise turmeric steroid saponin, glucose, sodium dodecyl sulfate, Bacillus belgii, Bacillus subtilis, white rot fungi and deionized water;

s3, adding 22mL of aspergillus niger suspension into 12.4L of the mixed solution, carrying out sealed fermentation for 4.5d at 30 ℃, adding 65mL of primary deionized water every 5 times in the fermentation process to obtain secondary fermentation solution, sterilizing the secondary fermentation solution at 90 ℃, and filtering thalli through an ultrafiltration membrane to obtain a mixed solution 2, wherein the main components of the mixed solution 2 comprise spiro-type curcuma longa steroid saponin, gluconic acid, sodium dodecyl sulfate and deionized water;

s4, adding excessive iron powder into the mixed solution 2, heating at 55 ℃, stirring at 2250rpm to generate a reaction solution containing ferrous gluconate, sodium dodecyl sulfate and Fe²⁺The complex forms foam floating on the surface of the reaction liquid due to violent stirring, the foam on the surface of the reaction liquid is scraped, the excessive iron powder is removed by filtration, then the mixture is separated by a superoleophobic filter membrane to obtain turmeric steroid saponin coarse substance and ferrous gluconate solution, the turmeric steroid saponin coarse substance and the ferrous gluconate solution are respectively refined to obtain turmeric steroid saponin and ferrous gluconate, and the turmeric steroid saponin and the ferrous gluconate solution are weighed.

The total effective viable count of the Bacillus beleisi bacterial suspension is not less than 1.0 x 10⁶cfu/mL, the total effective viable count of the bacillus subtilis suspension is not less than 1.5 multiplied by 10⁶cfu/mL, the total effective viable count of the white rot fungus suspension is not less than 1.4 multiplied by 10⁶cfu/mL; the total effective viable count of the aspergillus niger suspension bacterial suspension is not less than 1.8 multiplied by 10⁶ cfu/mL。

The crude curcuma longa steroid saponin is recrystallized, dried and refined to obtain solid curcuma longa steroid saponin; and the ferrous gluconate solution is subjected to decoloration, crystallization, drying and refining to obtain a ferrous gluconate solid.

Example 3

A method for extracting steroid saponin from Curcuma rhizome comprises the following steps:

s1, inserting 2.2kg of stems 11 of fresh turmeric rhizomes into an insertion tube 126 fixed on a cleaning beater, starting a motor 125, enabling the turmeric rhizomes to do self circular motion, starting a spray washing device 6, cleaning the turmeric rhizomes, sterilizing at a high temperature of 100 ℃ for 3min, starting a telescopic rod 3, driving a circular blade 5 to cut the turmeric rhizomes from the stems, enabling the turmeric rhizomes to fall into the bottom of the cleaning beater, closing a stop valve 9, spraying 2.2L of deionized water, starting a chopping beater 7, and beating to obtain turmeric pulp;

s2, adding 12mL of Bacillus belgii bacterial suspension, 12mL of Bacillus subtilis bacterial suspension and 20mL of white rot fungi bacterial suspension into the 2.4L of turmeric slurry obtained in the step S1, ventilating and fermenting for 3d at 35 ℃, then adding 400g of sodium dodecyl sulfate into the fermentation liquor, increasing the dissolving capacity of the turmeric steroid saponin in water, stirring for 20min, then filtering the fermentation liquor, washing the filter residue with deionized water (50 mL multiplied by 3), and combining the fermentation filtrate and the washing liquor to obtain a mixed liquor 1, wherein the main components of the mixed liquor 1 comprise turmeric steroid saponin, glucose, sodium dodecyl sulfate, Bacillus belgii, Bacillus subtilis, white rot fungi and deionized water;

s3, adding 25mL of aspergillus niger suspension into 12.5L of the mixed solution, carrying out sealed fermentation for 5d at 32 ℃, adding 80mL of primary deionized water every 5 times in the fermentation process to obtain secondary fermentation liquor, sterilizing the secondary fermentation liquor at 100 ℃, and filtering thalli through an ultrafiltration membrane to obtain a mixed solution 2, wherein the main components of the mixed solution 2 comprise spiro-type curcuma longa steroid saponin, gluconic acid, sodium dodecyl sulfate and deionized water;

s4, adding excessive iron powder into the mixed solution 2, heating at 60 ℃, stirring at 2500rpm to generate a reaction solution containing ferrous gluconate, sodium dodecyl sulfate and Fe²⁺The complex forms foam floating on the surface of the reaction liquid due to violent stirring, the foam on the surface of the reaction liquid is scraped, the excessive iron powder is removed by filtration, then the mixture is separated by a superoleophobic filter membrane to obtain turmeric steroid saponin coarse substance and ferrous gluconate solution, the turmeric steroid saponin coarse substance and the ferrous gluconate solution are respectively refined to obtain turmeric steroid saponin and ferrous gluconate, and the turmeric steroid saponin and the ferrous gluconate solution are weighed.

The total effective viable count of the Bacillus beleisi bacterial suspension is not less than 1.0 x 10⁶cfu/mL, the total effective viable count of the bacillus subtilis suspension is not less than 1.5 multiplied by 10⁶cfu/mL, the total effective viable count of the white rot fungus suspension is not less than 1.4 multiplied by 10⁶cfu/mL; the total effective viable count of the aspergillus niger suspension bacterial suspension is not less than 1.8 multiplied by 10⁶ cfu/mL。

The crude curcuma longa steroid saponin is recrystallized, dried and refined to obtain solid curcuma longa steroid saponin; and the ferrous gluconate solution is subjected to decoloration, crystallization, drying and refining to obtain a ferrous gluconate solid.

Comparative example 1

The operation of step S1 in example 1 is changed to: adding 2kg of fresh rhizome of Curcuma rhizome into a blender, cleaning, sterilizing at 80 deg.C for 2min, cutting, placing into pulping machine, adding 2L deionized water, and pulping to obtain Curcuma rhizome slurry; the remaining steps were carried out as described in example 1 to obtain turmeric steroid saponin solids and ferrous gluconate, and weighed.

Comparative example 2

The operation of step S2 in example 1 is changed to: adding 10mL of Bacillus belgii suspension, 10mL of Bacillus subtilis suspension and 18mL of white-rot fungi suspension into the yellow ginger slurry obtained in the step S1, ventilating and fermenting for 2d at 30 ℃, washing filter residue with deionized water (50 mL multiplied by 3), and combining the fermentation filtrate and washing liquid; the remaining steps were carried out as described in example 1 to obtain turmeric steroid saponin solids and ferrous gluconate, and weighed.

Comparative example 3

The operations of steps S2 and S3 in example 1 are changed to: s2, adding 10mL of Bacillus belgii bacterial suspension, 10mL of Bacillus subtilis bacterial suspension, 18mL of white rot fungi bacterial suspension and 20mL of Aspergillus niger bacterial suspension into the 2.2L of turmeric slurry obtained in the step S1, ventilating and fermenting for 2d at 30 ℃, then adding 350g of sodium dodecyl sulfate into fermentation liquor, increasing the dissolving capacity of turmeric steroid saponin in water, stirring for 10min, then filtering the fermentation liquor, washing filter residue with deionized water (50 mL multiplied by 3), combining fermentation filtrate and washing liquor, adding 50mL of once deionized water every 5 times in the fermentation process to obtain secondary fermentation liquor, then placing the secondary fermentation liquor at 80 ℃ for sterilization, and filtering thalli to obtain mixed liquor 2; the steps S1 and S4 were unchanged and were performed as described in example 1 to obtain turmeric steroid saponin solids and ferrous gluconate, and weighed.

Comparative example 4

The operation of step S2 in example 1 is changed to: adding 10mL of bacillus subtilis suspension and 18mL of white-rot fungi suspension into 2.2L of turmeric slurry obtained in the step S1, ventilating and fermenting for 2d at 30 ℃, then adding 350g of sodium dodecyl sulfate into fermentation liquor, increasing the dissolving capacity of turmeric steroid saponin in water, stirring for 10min, then filtering the fermentation liquor, washing filter residues with deionized water (50 mL multiplied by 3), and combining fermentation filtrate and washing liquor to obtain mixed liquor 1; the remaining steps were carried out as described in example 1 to obtain turmeric steroid saponin solids and ferrous gluconate, and weighed.

Test example 1

Determining the content of the steroid saponins of the turmeric: placing 0.1g of curcumine steroid saponin in a 25mL test tube with a plug, adding 10mL of sulfuric acid aqueous solution (1 mol/L), mixing uniformly, placing in a water bath kettle, treating in 100 deg.C water bath for 5h, performing acid hydrolysis to obtain hydrolysate; when the hydrolysate is cooled to room temperature, adjusting the pH value to be neutral by using a sodium hydroxide solution, transferring the neutral hydrolysate into a 50mL centrifuge tube, centrifuging at 8000r/min for 10min, carefully pouring out the supernatant, adding 25mL methanol, carrying out ultrasonic treatment for 40min, transferring into a new 50mL centrifuge tube, centrifuging at 8000r/min for 10min, collecting the supernatant, namely the methanol extract of the turmeric saponin, measuring the volume, sampling and detecting the content of the turmeric saponin, and representing the content of the turmeric saponin through the content of the turmeric saponin. Determining the content of diosgenin by high performance liquid chromatography;

the content of the ferrous gluconate is determined by adopting a titration method: dissolving 0.1g of ferrous gluconate in 15mL of water and 1mol/L of sulfuric acid solution, adding zinc powder to seal the plug, standing for 20min until decolorization, filtering, washing with newly boiled cold water, adding an o-diazophenanthrene indicator solution, titrating with a cerium sulfate titration solution (0.1 mol/L) until the solution is changed from orange to green, recording the usage amount of the cerium sulfate titration solution, and calculating to obtain the product; data are collated to obtain table 1;

as can be seen from table 1, the cleaning beater is adopted for fresh rhizome of turmeric to perform the operations of cleaning, high-temperature sterilization, crushing and beating step by step, the quality of the obtained steroid saponin of turmeric is improved by 11%, and the quality of ferrous gluconate is also improved, because the cleaning beater completes the operations of cleaning, crushing and beating in the same space, the epidermis integrity of the rhizome of turmeric is protected, the invasion of surface microorganisms is prevented, the opportunity of external microorganisms entering is reduced, and the stability of saponin in the rhizome of turmeric is maintained to the maximum; the addition of the sodium dodecyl sulfate improves the quality of the obtained curcuma steroid saponin by 18 percent, the quality content of ferrous gluconate is not greatly improved, and the sodium dodecyl sulfate promotes the dispersion degree of the spirostane steroid saponin in a water phase; compared with one-time fermentation, the survival rate of added strains can be improved by two-time fermentation, so that the quality of the steroid saponin of the turmeric and the quality of the ferrous gluconate are improved, and the effective content is improved; the combined action of the bacillus beleisi, the bacillus subtilis and the white rot fungi disintegrates starch, cellulose, lignin and the like which wrap the turmeric steroid saponin, thereby fully releasing the turmeric steroid saponin.

While the invention has been described in further detail with reference to specific embodiments thereof, it is not intended that the invention be limited to the specific embodiments thereof; for those skilled in the art to which the present invention pertains and related technologies, the extension, operation method and data replacement should fall within the protection scope of the present invention based on the technical solution of the present invention.