CN111454382A - Method for simultaneously extracting inulin and total caffeoylquinic acid from stevia rebaudiana Bertoni roots - Google Patents
Method for simultaneously extracting inulin and total caffeoylquinic acid from stevia rebaudiana Bertoni roots Download PDFInfo
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- CN111454382A CN111454382A CN202010214218.2A CN202010214218A CN111454382A CN 111454382 A CN111454382 A CN 111454382A CN 202010214218 A CN202010214218 A CN 202010214218A CN 111454382 A CN111454382 A CN 111454382A
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- stevia rebaudiana
- solid
- inulin
- acid
- root
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- 235000006092 Stevia rebaudiana Nutrition 0.000 title claims abstract description 71
- 229920001202 Inulin Polymers 0.000 title claims abstract description 46
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Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The present invention provides a method for simultaneously extracting inulin and total caffeoylquinic acid from stevia rebaudiana roots, wherein a step selected from the group consisting of extraction, fermentation, extraction, enzyme treatment, and resin separation is used. The method extracts or ferments various high value-added products by using stevia rebaudiana Bertoni root waste materials in the processes of planting and producing stevioside, and has good economic benefit, environmental benefit and social benefit.
Description
Technical Field
The invention belongs to the field of plant deep processing of plant extract production, and particularly provides a method for simultaneously extracting inulin and total caffeoylquinic acid from stevia rebaudiana Bertoni roots
Background
Inulin (also called inulin) is polysaccharide composed of fructose, has low calorie and no toxicity, and can be used for preventing dental caries, regulating lipid metabolism, promoting calcium absorption, reducing serum cholesterol, inhibiting cancer, and improving immunity as Bacillus bifidus proliferation factor: the water-soluble dietary fiber can be used as rheology and texture modifier, water retention agent, etc. Widely used in low calorie beverages, low fat or non-fat spreads, yogurt, ice cream, chocolate and the like (Klont R, Mannion P. insulin & oligofructose [ J ]. World of ingredients.1999 (9): 34-36).
Inulin is widely distributed in nature, and contains inulin in many plants, including Compositae (such as Jerusalem artichoke, herba Cichorii and dahlia), Juglandaceae, Gentianaceae, Asclepiadaceae, Saxifragaceae, Semiaquilegiae, Primulaceae, Boraginaceae, etc. in dicotyledonous plants, and Liliaceae (such as folium Allii tuberosi, Bulbus Allii Cepae, Bulbus Allii and Germinatus Phragmitis) and Gramineae in monocotyledonous plants. Inulin is most commonly commercially extracted from Jerusalem artichoke (commonly known as Jerusalem artichoke, with a 14% -19% inulin content in the root tuber) and chicory (with a 15% -20% inulin content in the root).
The jerusalem artichoke is a perennial herb of the compositae, has wide distribution, strong adaptability, low requirement on soil, barren and drought resistance, strong reproductive capacity, sand resistance and less plant diseases and insect pests, can be continuously used for many years after being planted once, and is mainly used for pickling pickles in China.
The average yield per mu of the jerusalem artichoke can reach 1200-5000kg (Zengxiaoyu, Loden forest, male Liu, Liujian. inulin, research status and development prospect [ J ] Chinese food additive, 2010(04):222-227), the inulin content of tubers of the jerusalem artichoke is very high and can account for 20% of the wet weight at most (Huqin, Chuaipeng, Wangjiamei, Liuqiang, Jinqiang, Yangyao, Wangbao. the planting and development of the jerusalem artichoke utilize [ J ] biomass chemical engineering, 2012,46(01): 51-54).
Chicory is a two-year-old herbaceous plant, is suitable for growing in marine climatic conditions, and is a commonly-planted vegetable variety in western europe (Zengxiaoyu, Loden forest, Liu Sheng Man, Liu Jian Xue. the research status and development prospect of inulin [ J ]. Chinese food additive, 2010(04): 222-. After chicory planting is optimized by water and fertilizer conditions, the annual yield is 79.3 to 199.7 tons per hectare, (yeanyeling, Liuyang, Weishijun, Poplar and Yaohangyan. the influence of different water and fertilizer treatments on the yield and the quality of chicory [ J ] the grass science, 2018,35(5):1161 and 1169.) the content of chicory polysaccharide in roots is 18 to 20% (Mary, Xuehun, Zhujinfang, Shenshifang. the determination and research of the content of chicory polysaccharide in chicory roots [ J ] the Chinese medicine guide report, 2019,25(10):63 to 65).
Stevia rebaudiana (Stevia rebaudiana Bertoni) belonging to Compositae family perennial grasses, originated in the region of the junction between Brazil and Paraguay, the leaves of which contain steviol glycoside sweet substances, have been widely used in the food industry as natural sweeteners (Dudao, Wufufu, Yi, Shang guan Xin, Cheng Guang, Peng Da Yong. response surface method for optimizing the extraction of 3, 5-dicaffeoylquinic acid from the hairy root of Stevia rebaudianaProcess [ J]The stevia rebaudiana is cultivated in large scale at home due to the wide application of stevioside, but only leaves are used for extracting the stevioside in the industrial production, so that the roots of the stevia rebaudiana are discarded as wastes, and the quality of the roots accounts for about 1/3 of the total amount of plants.
Regina A.C.
Arildo J.B.de Oliveira，Isolation and characterization of inulin with a high degree of polymerizationfrom roots of Stevia rebaudiana(Bert.)Bertoni[J]Carbohydrate Research,2015,411). In the study of compounds such as chlorogenic acid and fructooligosaccharide in stevia rebaudiana, it has been found that a large amount of phenolic acids such as 3, 5-dicaffeoylquinnic acid and 4, 5-dicaffeoylquinnic acid are contained in stevia rebaudiana (research on establishment of hairy root culture system of stevia rebaudiana and accumulation of chlorogenic acid compounds [ D]Jiangxi university of agriculture, 2015; chemical characteristics and predictive activity of microorganisms from Stevia rebaudiana (Bertoni) roots and in vitro acquired roots [ J]Carbohydratedepolymers.2016, 152: 718-. On the basis of the researches, the stevia rebaudiana, particularly the root of the stevia rebaudiana is more fully utilized, and products such as inulin, total caffeoylquinic acid and the like extracted from the stevia rebaudiana have good economic and environmental values.
The method extracts and measures the inulin amount and the total caffeoylquinic acid at the roots of the stevia rebaudiana by using a water extraction, alcohol extraction and biological enzymolysis method, and finds that the inulin content is rich (12% -30%), and the total caffeoylquinic acid amount is 2% -5%. Therefore, the stevia rebaudiana Bertoni root which is a waste material of the stevia sugar industry can be used as a raw material for extracting high-value inulin and phenolic acid. In addition, inulase (Zhang Tianxiang, Ding Ning, Yanchun Chun Guang, Sun Xiu, Wang Xin, Ma Jun Yan. inulase research progress [ J ]. Chinese brewing 2016,35(11): 21-25; Zhang super, Wang Jing, Tang Bo, Li Qijian. synchronous saccharification and fermentation Jerusalem artichoke for producing alcohol [ J ]. bioprocessing 2016,14(2):12-16), or esterase for hydrolyzing caffeoylquinic acid, or microorganism with the enzyme can be used for fermenting or enzymolyzing sweet Jerusalem artichoke root or extract to produce products such as alcohol, fructo-oligosaccharide, caffeic acid, and the like, thereby further enriching the product types and improving the economic benefit.
The implementation of the invention fully utilizes the waste materials in the stevia industry process, saves resources, and the stevia rebaudiana Bertoni root is waste, and the raw materials can greatly reduce the production cost of products such as inulin, total caffeoylquinic acid and the like; the purpose of simultaneously producing two or more products with high added values is achieved, and the advantages are outstanding.
Disclosure of Invention
Biomass is a main component in agricultural wastes, forestry wastes, food industry, wastes and domestic wastes, and a large amount of agricultural wastes are produced every year in China (liuyueqin, liuqingpu. agricultural waste resource utilization analysis expecting [ J ] agricultural machinery use and maintenance, 2013(11):109), the stevia rebaudiana industry, namely, a large amount of stevia rebaudiana roots are discarded when stevia rebaudiana leaves are used for producing stevioside, and the stevia rebaudiana is not fully applied, in the waste utilization research of the stevia rebaudiana roots, reducing sugar is researched, particularly, the reducing sugar is tested by using a test method and an HP L C test method, the inulin quantity and the total caffeoylquinic acid at the stevia rebaudiana roots are tested, the inulin content is found to be rich and reaches 12% -30%, the total caffeoylquinic acid quantity is 2% -5%, therefore, the stevia rebaudiana roots which can be used as waste materials of the stevia rebaudiana industries as raw materials, and can be used for producing alcohol or other fermentation products (Zhang-Chun, Zhang-Guo Henrong, Zhang-Guo, Jew, China Korea, China Sw.
In one aspect, the invention provides a method for extracting inulin and total caffeoylquinic acid, which is characterized in that the extraction method takes stevia rebaudiana Bertoni roots as raw materials.
Further, the method of the present invention comprises the step of adding water or other solvent to the stevia roots and subjecting to ultrasonic, heat or microwave treatment.
Further, the method comprises the step of leaching with water, hot water, ethanol and methanol.
Further, the solid-to-liquid ratio in the leaching step of the method is 1: 5-60.
Further, the method of the invention comprises a step of separating inulin or total caffeoylquinic acid by ethyl acetate extraction, macroporous resin adsorption or filtration
Further, the process of the present invention comprises a fermentation step.
Further, rhizopus, aspergillus niger, aspergillus oryzae, rhizopus oryzae, lactobacillus or yeast are used in the fermentation step of the method of the present invention.
Further, the method of the present invention comprises a step of treatment with inulinase or esterase.
Further, the method of the invention comprises the following steps: removing impurities from stevia rebaudiana Bertoni root, cleaning, leaching with 20-40% methanol at 50-70 deg.C, and mixing the solid and liquid ratios of 1:5-60, extracting and separating by ethyl acetate after solid-liquid separation to obtain total caffeoylquinic acid, and filtering and finishing raffinate to obtain inulin. .
Preferably, the stevia rebaudiana roots are taken to remove impurities and washed, and are leached by adopting methanol with the temperature of 60 ℃ and the concentration of 30 percent, and the solid-to-liquid ratio is 1: and 10, carrying out solid-liquid separation, extracting and separating by using ethyl acetate to obtain total caffeoylquinic acid, and filtering and finely processing raffinate to obtain inulin.
Further, the method of the invention comprises the following steps: removing impurities from stevia rebaudiana Bertoni root, cleaning, and leaching with water or 40-50 deg.C hot water at a solid-to-liquid ratio of 1: 10-50, performing solid-liquid separation to obtain stevia rebaudiana root leaching liquor, sterilizing, cooling, inoculating aspergillus niger, fermenting the leaching liquor, and separating to obtain fructo-oligosaccharide, caffeic acid and quinic acid.
Preferably, the stevia rebaudiana roots are taken to remove impurities and washed, and are leached by hot water at the temperature of 40 ℃, and the solid-to-liquid ratio is 1:50, performing solid-liquid separation to obtain stevia rebaudiana root leaching solution, sterilizing, cooling, inoculating Aspergillus niger, fermenting the leaching solution at 30 ℃ and pH of 5.0, and separating to obtain fructo-oligosaccharide, caffeic acid and quinic acid.
Further, the method of the present invention further comprises the step of using the processing residue as a feed or fertilizer.
Further, inulin and/or total caffeoylquinnic acid produced by the method of the present invention may be used to produce fructooligosaccharides or fructose.
Further, the method of the present invention can be used for preparing alcohol.
The Stevia rebaudiana Bertoni roots in the present invention include roots of various specific varieties of Stevia rebaudiana Bertoni, and are not limited to specific production areas and varieties. The stevia roots in the invention are not limited to the waste of stevioside extraction, but also include root waste in other related stevia industrial production or specially collected stevia roots.
In the present invention, various methods such as extraction, resin adsorption, or filtration can be selected depending on the properties of the product and the raw material, and the method is not limited to the examples in this section.
The fermentation strain of the present invention may be any of various naturally or artificially modified fungi, yeasts, bacteria, etc., which are known to be useful for fermentation, and is not limited to the species listed in this section.
Detailed Description
Example 1
Removing impurities from 100kg of stevia rebaudiana roots, cleaning, extracting the stevia rebaudiana roots directly or cutting the stevia rebaudiana roots into pieces by using normal-temperature water, wherein the solid-to-liquid ratio is 1: 30, obtaining stevia rebaudiana root leaching liquor after solid-liquid separation, and obtaining extracting liquor containing 15.7kg of inulin and 3.2kg of total caffeoylquinic acid through separation and extraction.
Removing impurities from the obtained stevia rebaudiana root leaching liquor, extracting and separating with ethyl acetate to obtain 2.5kg of total caffeoylquinic acid, and filtering the raffinate to obtain 14.5kg of inulin.
Example 2
Removing impurities from 100kg of stevia rebaudiana Bertoni root, cleaning, extracting the original or cut stevia rebaudiana Bertoni root with 40% ethanol at 30 deg.C: the solid-liquid ratio of the solvent is 1: 60, obtaining stevia rebaudiana root leaching liquor after solid-liquid separation, obtaining 10.3kg of inulin through fine filtration and separation, and obtaining 4.2kg of total caffeoylquinic acid through macroporous resin adsorption and separation.
Example 3
Removing impurities from 100kg of stevia rebaudiana Bertoni root, cleaning, extracting the original or cut stevia rebaudiana Bertoni root with 30% methanol at 60 deg.C, extracting the stevia rebaudiana Bertoni root: the solid-liquid ratio of the solvent is 1: 10, obtaining stevia rebaudiana root leaching liquor after solid-liquid separation, and simultaneously obtaining 15.9kg of inulin and 3.3kg of total caffeoylquinic acid through separation and extraction.
Example 4
Removing impurities from 100kg of stevia rebaudiana Bertoni root, cleaning, extracting with 40 deg.C hot water, and mixing the solid with the liquid at a solid-to-liquid ratio of 1:50, obtaining stevia rebaudiana root leaching liquor after solid-liquid separation, inoculating aspergillus niger after sterilization and cooling, fermenting the leaching liquor at the temperature of 30 ℃ and the pH value of 5.0, and obtaining 9.8kg of fructo-oligosaccharide, 1.1kg of caffeic acid and 0.83kg of quinic acid through separation.
Example 5
Removing impurities from 100kg of stevia rebaudiana Bertoni root, cleaning, extracting with 40 deg.C hot water, and mixing the solid with the liquid at a solid-to-liquid ratio of 1: 40, after solid-liquid separation, obtaining stevia rebaudiana root leaching liquor, sterilizing and cooling, inoculating rhizopus, fermenting at 32 ℃ and under the condition of pH5, and separating to obtain mash containing 10.2kg of fructose and 5.8kg of ethanol.
Example 6
Removing impurities from 100kg of stevia rebaudiana Bertoni root, cleaning, and extracting with 60 deg.C hot water, wherein the stevia rebaudiana Bertoni root: the solid-liquid ratio of the solvent is 1:50, obtaining stevia rebaudiana root leaching liquor after solid-liquid separation, adsorbing and separating by polyamide resin, and purifying by ethyl acetate to obtain 2.2kg of total caffeoylquinic acid.
Example 7
Removing impurities from 100kg of stevia rebaudiana Bertoni root, cleaning, chopping, adding a proper amount of water, keeping the temperature at 60 ℃ for 1h, cooling to room temperature, wherein the solid-to-liquid ratio is 1: and 5, adding Aspergillus niger, performing solid culture at 35 ℃ and pH5 for 3d, and separating to obtain 4.2kg of fructo-oligosaccharide, 0.81kg of caffeic acid and 0.23kg of quinnic acid.
Example 8
Removing impurities from stevia rebaudiana Bertoni root 100kg, cleaning, chopping, adding Aspergillus niger, and culturing at 35 deg.C and pH5 for 3 d. Adding water into the obtained solid fermentation product, and stirring, wherein the solid-liquid ratio is 1: 30, adding saccharomyces cerevisiae for fermentation, separating to obtain 9.2kg of ethanol, 0.63kg of caffeic acid and 0.17kg of quintic acid, and further processing the remainder to be used as a biological fertilizer.
Example 9
Removing impurities from sweet stevia root 100kg, cleaning, cutting, adding Rhizopus oryzae, performing solid fermentation at 30 deg.C and pH5, and separating the obtained fermentation product to obtain L-lactic acid 8.19kg, or directly using the fermentation product as biological fermented feed.
Example 10
Removing impurities from sweet stevia root 100kg, cleaning, cutting, adding Rhizopus, culturing at 30 deg.C and pH5 for 2d, adding Saccharomyces cerevisiae, fermenting for 24h, separating to obtain ethanol 11.3g, caffeic acid 0.59g and quinnic acid 0.16g, and using the residue as feed.
Example 11
The solvent was methanol as described in example 1. Adding inulase into inulin and total caffeoylquinic acid obtained after methanol separation, converting at 35 deg.C and pH of 6.0 for 36 hr, and extracting by conventional method to obtain fructo-oligosaccharide 5.8 kg.
Example 12
The solvent was ethanol as described in example 1. The extraction liquid containing 14.0kg of inulin and 3.9kg of total caffeoylquinic acid is obtained by separation and extraction.
The obtained stevia rebaudiana root leaching liquor is subjected to impurity removal, ethyl acetate is used for extraction and separation to obtain 3.2kg of total caffeoylquinic acid, and the raffinate is filtered to obtain 12.8kg of inulin.
Example 13
The solvent was 40% ethanol as described in example 1. The extraction liquid containing 14.7kg of inulin and 3.6kg of total caffeoylquinic acid is obtained by separation and extraction.
Removing impurities from the obtained stevia rebaudiana root leaching liquor, extracting and separating with ethyl acetate to obtain 2.9kg of total caffeoylquinic acid, and filtering the raffinate to obtain 13.3kg of inulin.
Example 14
The solvent was 60% methanol as described in example 1. The extraction liquid containing 14.9kg of inulin and 3.9kg of total caffeoylquinic acid is obtained by separation and extraction.
Removing impurities from the obtained stevia rebaudiana root leaching liquor, extracting and separating by using ethyl acetate to obtain 3.1kg of total caffeoylquinic acid, and filtering the raffinate to obtain 13.5kg of inulin.
Example 15
As described in example 1, the solid to liquid ratio was 1: 50. The extract containing 16.1kg of inulin and 3.5kg of total caffeoylquinic acid is obtained by separation and extraction.
Removing impurities from the obtained stevia rebaudiana root leaching liquor, extracting and separating with ethyl acetate to obtain 2.7kg of total caffeoylquinic acid, and filtering the raffinate to obtain 14.7kg of inulin.
Example 16
As described in example 1, the solid to liquid ratio was 1: 20. The extraction liquid containing 15.7kg of inulin and 3.2kg of total caffeoylquinic acid is obtained by separation and extraction.
Removing impurities from the obtained stevia rebaudiana root leaching liquor, extracting and separating with ethyl acetate to obtain 2.5kg of total caffeoylquinic acid, and filtering the raffinate to obtain 14.5kg of inulin.
The above examples are merely for illustrative clarity and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (15)
1. A method for extracting inulin and total caffeoylquinic acid is characterized in that the extraction method takes stevia rebaudiana Bertoni root as raw material
2. The method according to claim 1, which comprises the step of adding water or other solvent to the stevia rebaudiana roots and subjecting to ultrasonic, heat or microwave treatment.
3. A process according to claim 1 or 2, which includes the step of leaching with water, hot water, ethanol, methanol.
4. A process according to claim 3, wherein the solid to liquid ratio in the leach step is from 1:5 to 60.
5. A method according to any one of claims 1 to 4, comprising the step of isolating inulin or total caffeoylquinamic acid by extraction with ethyl acetate, adsorption on a macroporous resin or filtration
6. A process according to any one of claims 1 to 5, which includes a fermentation step.
7. The method according to claim 6, wherein the fermentation step is carried out using rhizopus, aspergillus niger, aspergillus oryzae, rhizopus oryzae, lactic acid bacteria or yeast.
8. A method according to any one of claims 1 to 5, which includes the step of treatment with an inulinase or esterase.
9. The method according to claim 1, comprising the steps of: removing impurities from stevia rebaudiana Bertoni root, cleaning, leaching with 20-40% methanol at 50-70 deg.C, and mixing the solid and liquid ratios of 1:5-60, extracting and separating by ethyl acetate after solid-liquid separation to obtain total caffeoylquinic acid, and filtering and finishing raffinate to obtain inulin.
10. The method according to claim 9, comprising the steps of: removing impurities from stevia rebaudiana roots, cleaning, leaching with 30% methanol at 60 ℃, wherein the solid-to-liquid ratio is 1: and 10, carrying out solid-liquid separation, extracting and separating by using ethyl acetate to obtain total caffeoylquinic acid, and filtering and finely processing raffinate to obtain inulin.
11. The method according to claim 1, comprising the steps of: removing impurities from stevia rebaudiana Bertoni root, cleaning, and leaching with water or 40-50 deg.C hot water at a solid-to-liquid ratio of 1: 10-50, performing solid-liquid separation to obtain stevia rebaudiana root leaching liquor, sterilizing, cooling, inoculating aspergillus niger, fermenting the leaching liquor, and separating to obtain fructo-oligosaccharide, caffeic acid and quinic acid.
12. The method according to claim 11, comprising the steps of: removing impurities from stevia rebaudiana roots, cleaning, leaching with hot water at 40 ℃, wherein the solid-to-liquid ratio is 1:50, performing solid-liquid separation to obtain stevia rebaudiana root leaching solution, sterilizing, cooling, inoculating Aspergillus niger, fermenting the leaching solution at 30 ℃ and pH of 5.0, and separating to obtain fructo-oligosaccharide, caffeic acid and quinic acid.
13. A method according to any one of claims 9 to 12, further comprising using the processing residue as a feed or fertiliser.
14. Use of a process according to claims 1-7 for the preparation of fructooligosaccharides or fructose.
15. Use of the method according to claims 1-7 for the preparation of alcohol.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102617667A (en) * | 2012-03-05 | 2012-08-01 | 南京师范大学 | Method for simultaneously preparing total caffeoylquinic acid and stevioside by taking stevia as raw material |
| CN103421853A (en) * | 2013-07-30 | 2013-12-04 | 南京师范大学 | Method for bioconversion of chlorogenic acids in plants by adopting aspergillus aculeatus |
| CN110662432A (en) * | 2016-11-29 | 2020-01-07 | 谱赛科美国股份有限公司 | Food ingredient from stevia |
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- 2020-03-24 CN CN202010214218.2A patent/CN111454382A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102617667A (en) * | 2012-03-05 | 2012-08-01 | 南京师范大学 | Method for simultaneously preparing total caffeoylquinic acid and stevioside by taking stevia as raw material |
| CN103421853A (en) * | 2013-07-30 | 2013-12-04 | 南京师范大学 | Method for bioconversion of chlorogenic acids in plants by adopting aspergillus aculeatus |
| CN110662432A (en) * | 2016-11-29 | 2020-01-07 | 谱赛科美国股份有限公司 | Food ingredient from stevia |
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