CN112175024A - Method for preparing high-purity stachyose from rehmannia - Google Patents
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- UQZIYBXSHAGNOE-USOSMYMVSA-N Stachyose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO[C@@H]2[C@@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O2)O1 UQZIYBXSHAGNOE-USOSMYMVSA-N 0.000 title claims abstract description 47
- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 title claims abstract description 47
- 241000405414 Rehmannia Species 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 28
- 238000000855 fermentation Methods 0.000 claims abstract description 29
- 230000004151 fermentation Effects 0.000 claims abstract description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000000843 powder Substances 0.000 claims abstract description 25
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000004927 clay Substances 0.000 claims abstract description 14
- 238000001914 filtration Methods 0.000 claims abstract description 13
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- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 11
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 11
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- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000003729 cation exchange resin Substances 0.000 claims abstract description 8
- 238000011033 desalting Methods 0.000 claims abstract description 8
- 239000003957 anion exchange resin Substances 0.000 claims abstract description 7
- 238000001035 drying Methods 0.000 claims abstract description 7
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- 239000007788 liquid Substances 0.000 claims description 37
- 238000000605 extraction Methods 0.000 claims description 35
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- 239000000047 product Substances 0.000 claims description 16
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- 235000013305 food Nutrition 0.000 description 1
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- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
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- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
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- 238000002360 preparation method Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
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- 210000002784 stomach Anatomy 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
Abstract
The invention relates to a subcritical and microbial fermentation combined method for preparing high-purity stachyose from rehmannia, which comprises the steps of dynamically extracting crushed rehmannia powder by taking subcritical water as an extracting solution, inoculating aspergillus oryzae and lactobacillus plantarum into the extracting solution for fermentation, adding activated carbon powder and activated clay for decoloring, purifying by anion exchange resin, desalting by cation exchange resin, filtering by a microporous membrane, concentrating under reduced pressure, and drying. The method is environment-friendly and efficient, the purity of the obtained stachyose is more than 91%, and the yield reaches 80%.
Description
Technical Field
The invention belongs to the technical field of plant extraction, and particularly relates to a method for preparing high-purity stachyose from rehmannia by combining subcritical and microbial fermentation.
Background
Radix rehmanniae is dry root tuber of rehmannia glutinosa Libosch of Scrophulariaceae, is cold in nature and sweet in taste, and has effects of clearing heat and cooling blood, nourishing yin and promoting fluid production. The main saccharide component in the dried rehmannia root is oligosaccharide, wherein the content of stachyose is high, and modern researches show that the stachyose can remarkably promote the proliferation of beneficial bacteria such as bifidobacteria, lactobacilli and the like in intestinal tracts, but can not be directly absorbed and utilized by digestive juice of intestines and stomach, so that the environment of the digestive tracts is rapidly improved, and the balance of microecological flora is adjusted; and has the potential effects of enhancing hematopoietic function, reducing blood sugar, promoting immunity, and resisting tumor; has certain bioactivity for regulating intestinal flora imbalance and subclinical hepatic encephalopathy of liver cirrhosis rats, reducing endotoxin in blood plasma and protecting liver.
At present, stachyose products have wide application value and market prospect at home and abroad, and the stachyose is gradually applied to various fields of food, health-care products, medicines, cosmetics, feed and the like in China. The extraction method of stachyose comprises a solvent extraction method, an ultrasonic-assisted extraction method, a microbial fermentation method, an enzymolysis extraction method and the like, but stachyose products directly extracted from plant raw materials have the problems of low purity, complex purification process, high stachyose loss rate and the like, and the efficacy and application of stachyose are greatly limited. In order to find a novel extraction process which is more environment-friendly and efficient, the extraction and purification process of the rehmannia stachyose is improved, and subcritical hot water extraction and microbial fermentation are combined to assist in preparing the stachyose.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for preparing high-purity stachyose from rehmannia by using a subcritical and microbial fermentation method, the method is environment-friendly and efficient, the purity of the obtained stachyose is more than 91%, and the yield reaches 80%.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing high-purity stachyose from rehmanniae radix by subcritical and microbial fermentation comprises the following steps:
1) putting the ground rehmannia powder into a conventional subcritical water extraction device, and dynamically extracting by taking subcritical water as an extracting solution, wherein the extraction pressure is 3-15 MPa, the extraction temperature is 100-200 ℃, and the extraction time is 30-90 min;
2) after extraction, cooling the extracting solution to room temperature, inoculating aspergillus oryzae and lactobacillus plantarum into the extracting solution for fermentation, wherein the fermentation temperature is 25-30 ℃, and the fermentation time is 1-3 days;
3) after fermentation, filtering, heating the filtrate to 60-90 ℃, adding activated carbon powder and activated clay, preserving heat for 0.5-1.5 h for decoloring, then cooling to room temperature, and centrifuging to obtain supernatant liquid;
4) purifying the supernatant by using anion exchange resin at the flow rate of 1-3 m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing effluent and/or eluent;
5) desalting the feed liquid obtained in the step 4) by using cation exchange resin at the flow speed of 5-10 m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing effluent and/or eluent;
6) filtering the feed liquid obtained in the step 5) by using a microporous filter membrane, concentrating under reduced pressure at 70 ℃, and drying to obtain a stachyose product.
Specifically, in the step 1), the dry rehmannia root raw material is crushed to 10-30 meshes; the feed-liquid ratio of the rehmannia powder to water is 1 g: 6-20 ml.
Specifically, in the step 2), the inoculation amount of aspergillus oryzae is 0.005-0.03% of the weight of the rehmannia root powder, and the inoculation amount of lactobacillus plantarum is 0.005-0.03% of the weight of the rehmannia root powder.
Specifically, in the step 3), the adding ratio (m/v) of the activated carbon powder to the filtrate is 0.2-5 g: 100ml, the adding ratio (m/v) of the activated clay to the filtrate is 0.1-2 g: 100 ml. The activated carbon and the activated clay are common commercial products which can be directly purchased.
Specifically, the microporous filter membrane of 0.6-0.8um is selected in the step 6).
Subcritical hot water extraction is a novel green extraction technology in recent years, the technology is that water is heated to the boiling point of more than 100 ℃ and the critical point of less than 347 ℃, a water body still keeps a liquid state, and under the condition, the water is in a subcritical state, so that the dissolving capacity of active ingredients is greatly improved, and the effect of improving the extraction efficiency is achieved. The technical problems of low extraction rate and low purity of the stachyose are solved by utilizing microbial assisted fermentation, and a more environment-friendly and more efficient preparation method of the stachyose is also provided.
The method for preparing the rehmannia stachyose by combining the subcritical hot water extraction and the enzyme-assisted fermentation method provided by the invention has the advantages that the yield of the obtained rehmannia stachyose can reach 81.54 percent at most, and the purity is 95.20 percent. Through subcritical hot water extraction, the thermal movement of water molecules is increased, and the extraction time of the rehmannia stachyose is shortened; the conversion rate of the stachyose is improved and the purity of the product is improved through microbial assisted fermentation; the taste of the finished product of the rehmannia stachyose is improved by desalting with cationic resin. The method does not use organic solvent, is green and environment-friendly, improves the extraction quality of stachyose, and expands the industrial value of rehmanniae radix.
Drawings
FIG. 1 is an HPLC chromatogram of a stachyose product obtained in example 1, and the result shows that the stachyose content is 93.02% and the total amount of sucrose and raffinose is 5.15%.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the following examples, but the scope of the present invention is not limited thereto.
In the invention, the raw materials are all common commercial products.
In the following examples, the anion exchange resin was 717 type anion resin; the cation exchange resin is 001 type cation resin which is a common commercial product;
aspergillus oryzae (Aspergillus oryzae) purchased from North Naita, product number BNCC 159241; lactobacillus plantarum (Lactobacillus plantarum) was purchased from North Naita, product number BNCC 193241.
Example 1
A method for preparing high-purity stachyose from rehmanniae radix by subcritical and microbial fermentation comprises the following steps:
1) crushing: crushing 100g of dry rehmannia root raw material to 10-30 meshes;
2) extraction: dynamically extracting the rehmannia powder obtained in the step 1) by taking subcritical water as an extracting solution, wherein the material-liquid ratio of the rehmannia powder to water is 1 g: 6ml, the extraction time is 30min, the extraction pressure is 5MPa, and the extraction temperature is 120 ℃;
3) after extraction, cooling the extract to room temperature, inoculating Aspergillus oryzae with a weight of 0.005% of the rehmannia powder and Lactobacillus plantarum with a weight of 0.01% of the rehmannia powder into the extract, and fermenting at 25 ℃ for 1.5 days;
4) after fermentation, filtering, heating the filtrate to 70 ℃, adding activated carbon powder and activated clay, keeping the temperature for 0.5h for decoloring, then cooling to room temperature, and centrifuging to obtain supernatant liquid; the adding ratio of the activated carbon powder to the filtrate is 0.5 g: 100ml, the adding ratio of the activated clay to the filtrate is 0.5 g: 100 ml;
5) purifying the supernatant with anion exchange resin at a flow rate of 3m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
6) desalting the feed liquid obtained in the step 5) by cation exchange resin at a flow rate of 10m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
7) filtering the feed liquid obtained in the step 6) by using a 0.6-0.8um microporous filter membrane, concentrating under reduced pressure at 70 ℃, and drying to obtain 80.41g of stachyose product.
The yield of stachyose prepared by the method is 80.41%, and the purity is 93.02%.
Example 2
A method for preparing high-purity stachyose from rehmanniae radix by subcritical and microbial fermentation comprises the following steps:
1) crushing: crushing 100g of dry rehmannia root raw material to 10-30 meshes;
2) extraction: dynamically extracting the rehmannia powder obtained in the step 1) by taking subcritical water as an extracting solution, wherein the material-liquid ratio of the rehmannia powder to water is 1 g: 10ml, the extraction time is 60min, the extraction pressure is 10MPa, and the extraction temperature is 150 ℃;
3) after extraction, cooling the extract to room temperature, inoculating Aspergillus oryzae 0.02 wt% of rehmanniae radix powder and Lactobacillus plantarum 0.02 wt% of rehmanniae radix powder into the extract, and fermenting at 30 deg.C for 3 days;
4) after fermentation, filtering, heating the filtrate to 80 ℃, adding activated carbon powder and activated clay, preserving heat for 1h for decolorization, then cooling to room temperature, and centrifuging to obtain supernatant liquid; the adding proportion of the activated carbon powder to the filtrate is 2 g: 100ml, the adding proportion of the activated clay to the filtrate is 2 g: 100 ml;
5) purifying the supernatant with anion exchange resin at a flow rate of 1m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
6) desalting the feed liquid obtained in the step 5) by using cation exchange resin at the flow speed of 5m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
7) filtering the feed liquid obtained in the step 6) by using a 0.6-0.8um microporous filter membrane, concentrating under reduced pressure at 70 ℃, and drying to obtain 79.74g of stachyose product.
The yield of stachyose prepared by the method is 79.74%, and the purity is 92.48%.
Example 3
A method for preparing high-purity stachyose from rehmanniae radix by subcritical and microbial fermentation comprises the following steps:
1) crushing: crushing 100g of dry rehmannia root raw material to 10-30 meshes;
2) extraction: dynamically extracting the rehmannia powder obtained in the step 1) by taking subcritical water as an extracting solution, wherein the material-liquid ratio of the rehmannia powder to water is 1 g: 15ml, the extraction time is 30min, the extraction pressure is 15MPa, and the extraction temperature is 180 ℃;
3) after extraction, cooling the extract to room temperature, inoculating Aspergillus oryzae with a weight of 0.01% of the rehmannia powder and Lactobacillus plantarum with a weight of 0.01% of the rehmannia powder into the extract, and fermenting at 30 ℃ for 2 days;
4) after fermentation, filtering, heating the filtrate to 90 ℃, adding activated carbon powder and activated clay, keeping the temperature for 1.5h for decolorization, then cooling to room temperature, and centrifuging to obtain supernatant liquid; the adding proportion of the activated carbon powder to the filtrate is 1 g: 100ml, the adding proportion of the activated clay to the filtrate is 1 g: 100 ml;
5) purifying the supernatant with anion exchange resinAt a flow rate of 2m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
6) desalting the feed liquid obtained in the step 5) by cation exchange resin at the flow rate of 8m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
7) filtering the feed liquid obtained in the step 6) by using a 0.6-0.8um microporous filter membrane, concentrating under reduced pressure at 70 ℃, and drying to obtain 81.54g of stachyose product.
The yield of stachyose prepared by the method is 81.54%, and the purity is 95.20%.
Example 4
A method for preparing high-purity stachyose from rehmanniae radix by subcritical and microbial fermentation comprises the following steps:
1) crushing: crushing 100g of dry rehmannia root raw material to 10-30 meshes;
2) extraction: dynamically extracting the rehmannia powder obtained in the step 1) by taking subcritical water as an extracting solution, wherein the material-liquid ratio of the rehmannia powder to water is 1 g: 10ml, and the extraction time is 90 min; the extraction pressure is 5MPa, and the extraction temperature is 200 ℃;
3) after extraction, cooling the extract to room temperature, inoculating aspergillus oryzae with the weight of 0.03% of that of the rehmannia powder and lactobacillus plantarum with the weight of 0.03% of that of the rehmannia powder into the extract for fermentation, wherein the fermentation temperature is 25 ℃, and the fermentation time is 2 days;
4) after fermentation, filtering, heating the filtrate to 60 ℃, adding activated carbon powder and activated clay, keeping the temperature for 1h for decolorization, then cooling to room temperature, and centrifuging to obtain supernatant liquid; the adding proportion of the activated carbon powder to the filtrate is 5 g: 100ml, the adding proportion of the activated clay to the filtrate is 2 g: 100 ml;
5) purifying the supernatant with anion exchange resin at a flow rate of 2m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
6) desalting the feed liquid obtained in the step 5) through cation exchange resin,flow velocity of 10m3Resin/h, collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
7) filtering the feed liquid obtained in the step 6) by using a 0.6-0.8um microporous filter membrane, concentrating under reduced pressure at 70 ℃, and drying to obtain 78.27g of stachyose product.
The yield of stachyose prepared by the method is 78.27%, and the purity is 91.86%.
Claims (5)
1. A method for preparing high-purity stachyose from rehmannia is characterized by comprising the following steps:
1) dynamically extracting the crushed rehmannia root powder by taking subcritical water as an extracting solution, wherein the extracting pressure is 3-15 MPa, the extracting temperature is 100-200 ℃, and the extracting time is 30-90 min;
2) after extraction, cooling the extracting solution to room temperature, inoculating aspergillus oryzae and lactobacillus plantarum into the extracting solution for fermentation, wherein the fermentation temperature is 25-30 ℃, and the fermentation time is 1-3 days;
3) after fermentation, filtering, heating the filtrate to 60-90 ℃, adding activated carbon powder and activated clay, preserving heat for 0.5-1.5 h for decoloring, then cooling to room temperature, and centrifuging to obtain supernatant liquid;
4) purifying the supernatant with anion exchange resin, and collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
5) desalting the feed liquid obtained in the step 4) by using cation exchange resin, and collecting effluent liquid; adding deionized water for elution, collecting eluent, and mixing the effluent and the eluent;
6) filtering the feed liquid obtained in the step 5) by using a microporous filter membrane, concentrating under reduced pressure, and drying to obtain a stachyose product.
2. The method for preparing high purity stachyose from rehmannia root as claimed in claim 1, wherein in step 1), the dry rehmannia root raw material is pulverized into 10-30 mesh; the feed-liquid ratio of the rehmannia powder to water is 1 g: 6-20 ml.
3. The method for preparing high purity stachyose from rehmannia glutinosa as claimed in claim 1 or 2, wherein in step 2), the amount of Aspergillus oryzae is 0.005-0.03% by weight of rehmannia glutinosa powder, and the amount of Lactobacillus plantarum is 0.005-0.03% by weight of rehmannia glutinosa powder.
4. The method for preparing high-purity stachyose from rehmannia as claimed in claim 3, wherein in the step 3), the adding ratio of the activated carbon powder to the filtrate is 0.2-5 g: 100ml, wherein the adding proportion of the activated clay to the filtrate is 0.1-2 g: 100 ml.
5. The method for preparing high purity stachyose from rehmanniae radix as claimed in claim 4, wherein 0.6-0.8um microporous membrane is selected in step 6).
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113974041A (en) * | 2021-12-02 | 2022-01-28 | 焦作市米奇食品饮料有限公司 | Preparation method of rehmannia beverage containing high rehmannia oligosaccharide |
| CN114213475A (en) * | 2021-12-29 | 2022-03-22 | 山东百龙创园生物科技股份有限公司 | Preparation method of stachyose |
| CN114249778A (en) * | 2022-01-05 | 2022-03-29 | 郑州铭泽生物科技有限公司 | Method for extracting stachyose from rehmannia and resource treatment method for extracting waste residues |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113974041A (en) * | 2021-12-02 | 2022-01-28 | 焦作市米奇食品饮料有限公司 | Preparation method of rehmannia beverage containing high rehmannia oligosaccharide |
| CN113974041B (en) * | 2021-12-02 | 2024-02-06 | 焦作市米奇食品饮料有限公司 | Preparation method of rehmannia beverage containing high rehmannia oligosaccharide |
| CN114213475A (en) * | 2021-12-29 | 2022-03-22 | 山东百龙创园生物科技股份有限公司 | Preparation method of stachyose |
| CN114213475B (en) * | 2021-12-29 | 2023-01-31 | 山东百龙创园生物科技股份有限公司 | Preparation method of stachyose |
| CN114249778A (en) * | 2022-01-05 | 2022-03-29 | 郑州铭泽生物科技有限公司 | Method for extracting stachyose from rehmannia and resource treatment method for extracting waste residues |
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