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    • 专利摘要:
    The disclosure provides a flour inulin of Helianthus tuberosus (L. 1753) L. for ameliorating functional dyspepsia and a method of preparation and use thereof, which belongs to the technical field of deep processing of natural products.
    公开号:BE1027863B1
    申请号:E20205421
    申请日:2020-06-11
    公开日:2021-07-14
    发明作者:Ming Zhang；Tingting Gao；Lixin Wei；Hongtao Bi；Cen Li；Hongxia Yang；Yuzhi Du
    申请人:Northwest Inst Plateau Bio Cas；
    IPC主号:
    专利说明:

    A FLOUR INULIN FROM HELIANTHUS TUBEROSUS (L. 1753) L. FOR IMPROVE FUNCTIONAL DYSPEPSIA AND A PROCESS OF PREPARATION AND USE OF IT
    TECHNICAL FIELD The disclosure belongs to the technical field of deep processing of natural products, and specifically relates to a flour inulin of Helianthus tuberosus {L. 1753) L. for ameliorating functional dyspepsia and a method of preparation and use thereof.
    BACKGROUND Inulin has been widely concerned due to the fact that it has antioxidant activity, that it is able to improve the diversity of the intestinal flora, promote intestinal peristalsis, and improve motility disorders. gastrointestinal. In the past 10 years, the demand for inulin in the Chinese market has grown at an average annual growth rate of around 15%, until 2018, the demand for inulin in the Chinese market was around 8,000 tons. , to achieve a growth of about 10 times. Overall, it was shown from the 2015 data that the demand for inulin in the world market was around 0.2 million tons, new products containing inulin accounted for around 24% of new products based on inulin. of dietary fiber globally, ranking first. The raw materials of [inulin are very different between China and European and American countries, Helianthus tuberosus L. is used as a raw material for industrial production of inulin in Europe, while in industrial production of inulin in our country, the raw material is Helianthus tuberosus (L. 1753} L. (Wei Lingyun, et al, 2008). Compared to European countries, studies on inulin of Helianthus tuberosus (L. 1753) L. in our country have started late, and currently hot water extraction is used to obtain inulin from Helianthus tuberosus (L. 1753) L., about 650 kg of deposits from Helianthus tuberosus (L. 1753) L. have been obtained after the hot calf extraction of 1 ton of fresh Helianthus tuberosus (L. 1753) L., the deposits of Helianthus tuberosus (L. 1753) L. were dried to obtain a flour of Helianthus tuberosus {L. 1753) L ., which contained a large amount of saccharide compounds te ls as pectin, inulin, cellulose / hemicellulose. Therefore, it was expected that a thorough extraction and use of saccharide compounds in flour of Helianthus tuberosus (L. 1753) L. would increase the availability of Helianthus tuberosus (L. 1753) L. in domestic enterprises. and promote production value without changing the inherent process conditions of inulin production.
    Currently, researchers have started to focus on the efficient use of flour from Helianthus tuberosus (L. 1753) L., reports have been published on studies of reuse of pectin and monosaccharides in flour of Helianthus tuberosus (L. 1753) L., while no report has been published on the in-depth use of inulin in flour of Helianthus tuberosus (L. 1753) L.
    Functional dyspepsia (FD) is one of the most common functional gastrointestinal illnesses.
    Due to the recurrent attacks of this disease, and the fact that Western medicines in clinical use are currently most often accompanied by serious side effects of the digestive tract and nervous system, the exploration of new therapeutic drugs from natural products is has become [axis of current medical research for the treatment of this disease.
    Linulin is a natural dietary fiber soluble in water, being difficult to hydrolyze and digested by stomach acid, which can only be used by beneficial microorganisms in the colon, thus improving the intestinal environment.
    Du Zhaoping (2014) studied the effects of inulin on the in vitro proliferation of Lactobacillus casei and Lactobacillus plantarum with MRS liquid medium containing inulin, with results indicating that inulin could significantly promote both proliferations in vitro.
    Besides regulating the intestinal flora, inufin also has the effects of promoting intestinal peristalsis, and increasing gastrointestinal motility, thus preventing and improving constipation, diarrhea, colon cancer, etc.
    Li Suyan, et al. (2015) discussed the clinical effects of probiotics combined with inulin in the treatment of acute diarrhea in adults through experiments.
    Probiotics combined with inulin have been shown from the results to be useful in improving the functions of the intestinal mucous membranes of patients with acute diarrhea, promoting the reestablishment of the balance of the intestinal flora, with significant therapeutic effects. in the treatment of acute diarrhea in adults by synergistic actions of the two compounds, and being safe and reliable.
    However, no report has yet been published on the treatment of functional dyspepsia with inulin from Helianthus tuberosus (L. 1753) L. 35.
    SUMMARY In view of this, the object of the invention is to provide a flour inulin of Helianthus iuberosus (L. 1753) L. for ameliorating functional dyspepsia and a method of preparation and use thereof.
    The disclosure provides a method of preparing inulin from flour of Helianthus tuberosus (L. 1753) L. for ameliorating functional dyspepsia, which includes the following steps: 1) mixing powdered flour of Helianthus tuberosus (L. 1753) L with a buffer solution containing glycosidases, and enzymatic extraction, to give an extract; 2) ultrafiltration of the extract to remove macromolecular impurities such as proteins and pectin, to give a crude inulin extract of Helianthus tuberosus (L. 1753) L flour; 3) desalination and purification of the crude inulin extract from the flour of Helianthus tuberosus (L. 1753) L., to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L., desalinated; 4) ultrafiltration of the inulin extract from flour of Helianthus tuberosus (L. 1753) L. desalinated for the second time to remove small molecule impurities such as monosaccharides and oligosaccharides, drying, to give Pinulin from Helianthus flour tuberosus (L. 1753) L.
    Preferably, the buffer solution containing glycosidases in step 1) comprises, with a buffer solution of citric acid-sodium citrate at 0.01-0.5 mol! as solvent, the following glycosidases: pectinase, cellulase, hemicellulase and xylanase; The ratio of the mass of the flour powders of Helianthus tuberosus (L. 1753) L to the activities of pectinase, cellulase, hemicellulase and xylanase is, in order, 1 g: 3-20 U : 5-15 U: 10-30 U: 5-20 U.
    Preferably, the ratio of the mass of the flour powders of Helianthus tuberosus (L. 1753) L. to the volume of the buffer solution containing glycosidases in step 1) is 1 g: 10-30 ml; The enzymatic extraction temperature is 45-55 ° C, the enzymatic extraction time is 1-3 h.
    Preferably, the limiting molecular weight of the ultrafiltration membrane used in the ultrafiltration of step 2) is 20,000 Da, with a membrane area of 0.2 m2; the pressure differential in the ultrafiltration operation is 1.0 MPa, with a circular flow of 2 l / min.
    Preferably, the desalination and purification in step 3) are carried out with cation and anion exchange resins: The volume ratio of strongly acidic cation exchange resins: strongly basic anion exchange resins is 1: 1 -2; The ratio of the total volume of lonic resins to the volume of crude inulin extract from flour of Helianthus tuberosus (L. 1753) L. is 1: 11.5; The duration of desalination and purification is 60-90 min. Preferably, the molecular weight limit of the ultrafiltration membrane used in the ultrafiltration for the second time in step 3) is 1000 Da, with a membrane area of 0.2 m2, a pressure differential of service of 1.0 MPa, and a circular flow of 2 l / min; the drying includes spray drying; the inlet temperature of spray drying is 185-195 ° C, the outlet temperature is 85-95 ° C, with a feed rate of 800-1000 ml / h.
    The disclosure provides a Helianthus tuberosus (L. 1753) L. flour inulin for improving functional dyspepsia prepared by the above preparation method, the purity of Helianthus tuberosus (L. 1753) L. flour inulin. is = 91%: the content of total saccharides is 293%, in which the content of 1F-fructofuranosylnystose and of fructans of higher degrees of polymerization is z 75%; the uronic acid content is <0.34%: and the protein content is <0.61%.
    Preferably, the total antioxidant capacity of Helianthus tuberosus (L. 1753) L. flour inulin is expressed by TEAC; the TEAC is x milligrams of water soluble vitamin E in equivalent per gram of inulin sample from Helianthus tuberosus (L. 1753) L flour; The total antioxidant capacity of inulin from flour of Helianthus tuberosus (L. 1753) L. is 3.56-4.25 mTE / g. The disclosure provides a use of the flour inulin of Helianthus tuberosus (L.
    1753) L. in the preparation of medicaments for the treatment of functional dyspepsia.
    The disclosure provides a use of the flour inulin of Helianthus tuberosus (L.
    1753) L. in health care products to improve gastric contents residual and gastrointestinal hormone levels, gut flora disturbances and / or oxidative stress level.
    The method of preparing Helianthus tuberosus (L. 1753) L. flour inulin provided in this disclosure is as follows: mixing Helianthus tuberosus (L. 1753) L. flour powders with a buffer solution containing glycosidases, and enzymatic extraction, ultrafiltration of the extract to remove macromolecular impurities such as proteins and pectin, desalination and purification of the crude inulin extract from flour of Helianthus tuberosus {L. 1753) L. before ultrafiltration for the second time to remove small molecule impurities such as monosaccharides and oligosaccharides, drying, to give the flour inulin of Helianthus tuberosus (L. 1753) L.
    Compared to the method of preparing Pinulin from flour of Helianthus tuberosus (L. 1753) L. with the traditional hot water extraction, the preparation method provided in this disclosure exhibits as a characteristic feature a high extraction efficiency, the Inulin yield from Helianthus tuberosus (L. 1753) L. flour is increased by 24.58-39.74%, which increases the availability of raw materials from Helianthus tuberosus (L. 1753) L and promotes the value of production.
    On the other hand, the preparation process provided in this disclosure further limits the temperature used in the enzymatic extraction to 45-55 ° C, the extraction temperature was reduced compared to the traditional hot water extraction in which the temperature d. The extraction was above 80 ° C, which favors the reduction of energy consumption during extraction.
    The disclosure provides a Helianthus tuberosus (L. 1753) L. flour inulin for improving functional dyspepsia prepared by the above preparation method, the purity of Helianthus tuberosus (L. 1753) L. flour inulin. is = 91%: the content of total saccharides is 293%, in which the content of 1F-fructofuranosylnystose and of fructans of higher degrees of polymerization is 275%; the uronic acid content is <0.34%: and the protein content is <0.61%. Compared with traditional hot water extraction, the purity of Helianthus tuberosus (L. 1753) L. flour inulin is increased by 2.4% -5.2%, the total saccharide content is increased from 3.9% -7.7%, the content of 1F-fructofuranosylnystose and of fructans of higher degrees of polymerization in inulin is increased by 3.7% -6.9%. Furthermore, through the detection of the total antioxidant capacity of Helianthus tuberosus (L. 1753) L. flour inulin, which is expressed by TEAC, the total antioxidant capacity of Helianthus tuberosus flour inulin ( L. 1753) L. provided in this disclosure is 3.56-4.25 mgTE / g, the total antioxidant capacity (value
    TEAC) is increased by 45.85-55.73% compared to that of Finulin from Helianthus tuberosus (L. 1753) L. commercial. The disclosure provides a use of Helianthus tuberosus (L. 1753) L. flour inulin in the preparation of medicaments for the treatment of functional dyspepsia. I! It has been shown from experiments that the effects of Pinulin on improving the residual level of gastric contents and gastrointestinal hormone levels in ED rats had become greater: compared to inulin from Helianthus tuberosus (L. 1753) Commercial L., in the experimental group of the inulin prepared in this disclosure, the gastric emptying rate was increased by 9.47-18.39%, the MTL content was increased by 14.60-25.70 %, the Gréhline content was reduced by 1.22-2.43%, the Gas content was reduced by 4.21-7.62%, the 5-HT content was reduced by 3.84-5.46 %; at the same time, the effect of [inulin on improving disturbances of the intestinal flora in DF rats became more significant.
    DESCRIPTION OF EMBODIMENTS The disclosure provides a process for preparing Helianthus tuberosus (L. 1753) L. flour inulin to ameliorate functional dyspepsia, which includes the following steps: 1) mixing Helianthus tuberosus flour powders ( L. 1753) L. with a buffer solution containing glycosidases, and enzymatic extraction, to give an extract; 2) ultrafiltration of the extract to remove macromolecular impurities such as proteins and pectin, to give a crude inulin extract of Helianthus tuberosus (L. 1753) L flour; 3) desalination and purification of the crude inulin extract from the flour of Helianthus tuberosus (L. 1753) L., to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L., desalinated; 4) ultrafiltration of the inulin extract from the flour of Helianthus tuberosus {L. 1753) L. desalinated for the second time to remove small molecule impurities such as monosaccharides and oligosaccharides, drying, to give the flour inulin of Helianthus tuberosus (L. 1753) L.
    In this disclosure, an extract was obtained by mixing Helianthus tuberosus (L. 1753) L. flour powders with a buffer solution containing glycosidases, and enzymatic extraction.
    In this disclosure, the flour powders of Helianthus tuberosus (L. 1753) L. have preferably been obtained by grinding a flour of Helianthus tuberosus (L. 1753).
    L. produced during industrial production with a food grinder.
    The grain sizes of the flour powders were preferably 50-300 mesh, more preferably 150 mesh.
    There was no particular limitation on the model of the food grinder for grinding in the disclosure, as long as a food grinder well known in the art was employed.
    In embodiments of the disclosure, the food grinder was purchased from Chengxun Industrial Co., Ltd., with a model of CXP-DJ-S-5T.
    In this disclosure, the ratio of the mass of Helianthus tuberosus (L. 1753) L. flour powders to the volume of the buffer solution containing glycosidases is preferably 19: 10-30 ml, more preferably 19: 20 mi.
    The glycosidase-containing buffer solution comprises, preferably with a 0.01-0.5 mol / sodium citric acid-sodium citrate buffer solution, at pH 4.0-5.5, as solvent, the the following glycosidases: pectinase, cellulase, hemicellulase and xylanase; the ratio of the mass of the flour powders of Helianthus tuberosus (L. 1753) L. to the activities of pectinase, cellulase, hemicellulase and xylanase is, in order, preferably 1 g: 3-20 U: 5-15 U: 10-30 U: 5-20 U, more preferably 1g: 5-15 U: 8-12 U: 15-25 U: 8-15 U, more preferably between all of 19: 10 U: 10 U: 20 U: 12 U.
    The temperature of the enzymatic extraction is preferably 45-55 ° C, more preferably 50 ° C.
    The concentration of the citric acid-sodium citrate buffer solution is preferably 0.1-0.4 mol / l, more preferably 0.25 mol.
    The duration of the enzymatic extraction is preferably 1-3 h, more preferably 2 h.
    The enzymatic extraction process is beneficial for the separation of proteins and polysaccharides in Helianthus tuberosus (L. 1753) L. flour from their cells under the effects of various biological enzymes.
    After obtaining the extract, the extract was subjected to ultrafiltration in the disclosure to remove macromolecular impurities such as proteins and pectin, to give a crude extract of Helianthus tuberosus flour nulin {L. 1753) L.
    In this disclosure, the limiting molecular weight of the ultrafiltration membrane used in the ultrafiltration is preferably 20,000 Da, the membrane area is preferably 0.2 m2. There was no particular limitation on the equipment used for membrane separation and the resources of the ultrafiltration membrane, as long as the equipment used for the membrane separation was employed as well as the resources of the membrane. ultrafiltration which are well known in the art.
    In embodiments of the disclosure, the equipment used for membrane separation is preferably model RNF-0460. The ultrafiltration membrane was preferably purchased from Xiamen Filter & Membrane Technology Co., Lid.
    The pressure differential in the ufiltration operation is preferably 1.0 MPa, and the circular flow is preferably 2 l / min.
    The ultrafiltration process is capable of effectively removing macromolecular impurities such as protein and pectin.
    After obtaining the crude inulin extract of flour of Helianthus tuberosus (L. 1753) L., the crude inulin extract of flour of Helianthus tuberosus (L. 1753) L. was desalinated and purified in the disclosure. to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated. | In this disclosure, the desalination and purification have preferably been carried out with cation and anion exchange resins; The volume ratio of strongly acidic cation exchange resins: strongly basic anion exchange resins is 1: 1-2, more preferably 1: 1.5. The ratio of the total volume of ionic resins to the volume of the crude inulin extract of flour of Helianthus tuberosus (L. 1753) L. is 1: 11.5, more preferably 1: 1.2, La duration of desalination and purification is preferably 60-90 min, 70-80 min.
    There was no particular limitation on the desalination operations for cation and anion exchange resins, as long as the technical schemes for desalination with cation and anion exchange resins which are well known in the art were employed. art.
    After obtaining the inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated, [inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated was subjected to ultrafiltration for the second times to remove small molecule impurities such as monosaccharides and oligosaccharides and dried in the disclosure, to give the flour inulin of Helianthus tuberosus (L. 1753) L.
    In this disclosure, the limiting molecular weight of the ultrafiltration membrane used in the ultrafiltration for the second time is preferably 1000 Da, the membrane area is preferably 0.2 m2. There was no particular limitation on the equipment used for membrane separation as well as the resources of the ultrafiltration membrane in the disclosure, as long as the equipment used for the membrane separation was employed as well as the resources of the ultrafiltration membrane. the ultrafiltration membrane which were well known in the art.
    In embodiments of the disclosure, the equipment used for membrane separation is preferably model RNF-0460. The ultrafiltration membrane is preferably purchased from Xiamen Filter & Membrane Technology Co, Ltd.
    The working pressure differential is preferably 1.0 MPa, and the circular flow is preferably 2 l / min.
    The second ultrafiltration process is beneficial in increasing the removal rates of small molecule impurities such as monosaccharides and oligosaccharides. There was no particular limitation on the drying method in the disclosure, all inulin drying methods well known in the art could be employed.
    The drying is preferably spray drying; the inlet temperature of the spray drying is 185-195 ° C, more preferably 190 ° C; the outlet temperature is preferably 85-95 ° C, more preferably 90 ° C.
    The feed rate is preferably 800-1000ml / h, more preferably 900ml / h.
    Spray drying is beneficial in decreasing the quality loss of Finulin.
    The disclosure provides a Helianthus tuberosus (L. 1753) L. flour inulin for improving functional dyspepsia prepared by the above preparation method, the purity of Helianthus tuberosus (L. 1753) L. flour inulin. is = 91%: the determination method was available in document 1 (Xiao Zijun, et al.
    Studies on extraction processes of inulin in the Helianthus tuberosus (L. 1753) L. [J]. Modern Food Science and Technology, 2013, 29 (2), 315-318.). The total saccharide content in Helianthus tuberosus (L. 1753) L. flour inulin is = 93%, the total saccharide content is determined with fructose as standard monosaccharide, the determination method was available in the document 2 (Dubois M, Gilles KA, Hamilton JK, et al.
    Colorimetric method for determination of sugars and related substances [J]. Analytical Chemistry, 1956, 28: 350-356). In total saccharides, the content of 1F-fructofuranosylnystose and of fructans of higher degrees of polymerization is 275%; in fructans, the configuration of the fructose residue and glycosidic bond were predominantly —1) -BD-Fruf- (2— and BD-Fruf- (2->. The method of determination was available in document 3 (Fu Q, Liang T, Zhang X, et al.
    Carbohydrate separation by hydrophilic interaction liquid chromatography on a ‘click maltose columniJ]. Carbohydrate Research, 2010, 345: 2690-2697). The uronic acid content in [Helianthus tuberosus (L. 1753) L. flour inulin is <0.34%; the determination method was available in document 4 (Blumenkrantz N, Asboe-Hansen G.
    New method for quantitative determination of uronic acids. [J]. Analytical Biochemistry, 1973, 54: 484-489.). The protein content in the inulin of flour of Helianthus tuberosus (L. 1753) L. is <0.61%. The determination method was available in document 5 (Sedmark
    JJ, Grossberg SE. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250 [J]. Analytical Biochemistry, 1979, 79: 544-552).
    In this disclosure, the total antioxidant capacity of Helianthus tuberosus flour tinulin {L. 1753) L. is preferably expressed by TEAC: TEAC is x milligrams of water soluble vitamin E in equivalent per gram of inulin sample from Helianthus tuberosus (L. 1753) L flour; the total antioxidant capacity of Helianthus tuberosus flour inulin {L. 1753) L is preferably 3.56-4.25 mgTE / g. The total antioxidant capacity (TEAC value) was increased by 45.85-55.73% compared to that of commercial inulin (Orafit® HIS Inulin powder, batch number: RHWCJ8ACJ8).
    The disclosure provides a use of Helianthus tuberosus flour inulin (L.
    1753) L. in the preparation of medicaments for the treatment of functional dyspepsia.
    The disclosure provides a use of Helianthus tuberosus (L. 1753) L. flour inulin in health care products for improving the residual level of gastric contents and levels of gastrointestinal hormones, gastrointestinal disorders. intestinal flora and / or the level of oxidative stress.
    Helianthus tuberosus (L. 1753) L. flour inulin for ameliorating functional dyspepsia provided in this disclosure and a method of preparation and use thereof will be described in detail below in conjunction with embodiments, which cannot be interpreted as limiting the scope of the invention.
    Embodiment 1 (1) Flour of Helianthus tuberosus (L. 1753) L. produced during the industrial production of inulin from Helianthus tuberosus (L. 1753) L. was ground with a food mill (manufacturer: Chengxun Industrial Co., Ltd, model: CXP-DJ-S-5T); (2) At a solid-liquid ratio of 1:10 (g / ml), buffered saline (citric acid-sodium citrate buffer solution, 0.01 mol, pH 4.0) containing glycosidases ( the ratio of enzymes to substrates was respectively pectinase 3 U / g, cellulase 5 U / g, hemicellulase 10 U / g, and the proportion of xylanase was 5 U / 9), extraction at 45 ° C for 3 h: (3 ) The extract was subjected to ultrafiltration to remove macromolecular impurities such as protein and pectin (membrane separation equipment: model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 20,000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co., Lid .; the working pressure differential at
    1.0 MPa, with a circular flow of 2 l / min), to give a crude extract of inulin from the flour of Helianthus tuberosus (L. 1753) L .; (4) The crude inulin extract of Helianthus tuberosus (L. 1753) L. flour was desalinated and purified with cation and anion exchange resins (the volume ratio of strongly acidic cation exchange resins: resins strongly basic anion exchangers was 1: 1, the volume ratio of ionic resins to the crude inulin extract of flour of Helianthus tuberosus (L. 1753) L. was 1: 1, the resin treatment time was 60 min), to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated;
    (5) Inulin extract from Helianthus tuberosus (L. 1753} L. desalinated flour was subjected to ultrafiltration to remove small molecule impurities such as monosaccharides and oligosaccharides (membrane separation equipment: model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 1000 Da, with a membrane area of 0.2 m2, Xiamen
    Filter & Membrane Technology Co, Ltd. ; working pressure differential at 1.0 MPa, with a circular flow of 2 l / min), to give the inulin extract of Helianthus tuberosus flour {L. 1753) L .;
    (6) The purified inulin solution was spray dried (inlet temperature was 185 ° C, outlet temperature was 85 ° C, with a feed rate of
    800 ml / h), to give the Helianthus tuberosus flour inulin product {L. 1753) L.
    Embodiment 2 (1) Flour of Helianthus tuberosus (L. 1753) L. produced during industrial production of inulin from Helianthus tuberosus (L. 1753) L. was ground with a food mill (manufacturer: Chengxun Industrial Co., Ltd, model: CXP-DJ-S-5T);
    (2) At a solid-liquid ratio of 1:30 (g / mI), buffered saline (citric acid-sodium citrate buffer solution, 0.5 mol /, pH 5.5) containing glycosidases (the ratio of enzymes to substrates was respectively pectinase 20 U / g, cellulase 5 U / g, hemicellulase 30 U / g, and the proportion of xylanase was 20 U / g), extraction at 55 ° C for 1 h:
    (3) The extract was subjected to ultrafiltration to remove macromolecular impurities such as protein and pectin (membrane separation equipment: model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 20,000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co.
    Ltd. ; the operating pressure differential at
    1.0 MPa, with a circular flow of 2 l / min}, to give a crude inulin extract from the flour of Helianthus tuberosus (L. 1753) L;
    (4) The crude inulin extract of Helianthus tuberosus (L. 1753) L. flour was desalinated and purified with cation and anion exchange resins (the volume ratio of strongly acidic cation exchange resins: resins strongly basic anion exchangers was 1: 2, the volume ratio of ionic resins to the crude inulin extract of flour of Helianthus tuberosus (L. 1753) L. was 1: 1.5, the treatment time per resin was 90 min), to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated; (5) Inulin extract from Helianthus tuberosus (L. 1753) L. desalinated flour was subjected to uterfiltration to remove small molecule impurities such as monosaccharides and oligosaccharides (membrane separation equipment : Model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 1000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co., Ltd.; the working pressure differential at 1.0 MPa, with a circular flow of 2 l / min), to give the inulin extract of the flour of Helianthus tuberosus (L. 1753) L .; (6) The purified inulin solution was spray dried (the inlet temperature was 195 ° C, the outlet temperature was 95 ° C, with a feed rate of 1000 ml / h}, to give the inulin product of flour of Helianthus tuberosus (L. 1753) L.
    Embodiment 3 (1) Flour of Helianthus tuberosus (L. 1753) L. produced during industrial production of inulin from Helianthus tuberosus (L. 1753) L. was ground with a food mill (manufacturer: Chengxun Industrial Co, Ltd, model: CXP-DJ-S-5T); (2) At a solid-liquid ratio of 1:20 (g / ml), buffered saline (citric acid-sodium citrate buffer solution, 0.1 mol / l, pH 5) containing glycosidases ( the ratio of enzymes to substrates was respectively pectinase 15 U / g, cellulase 10 U / g, hemicellulase 20 U / g, and the proportion of xylanase was 15 U / g), extraction at 50 ° C for 2 h; (3) The extract was subjected to ultrafiltration to remove macromolecular impurities such as protein and pectin (membrane separation equipment: model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 20,000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co, Lid .; the working pressure differential at 1.0 MPa, with a circular flow of 2 min), to give a crude extract d Helianthus tuberosus (L. 1753) L flour inulin;
    (4) The crude inulin extract of Helianthus tuberosus (L. 1753) L. flour was desalinated and purified with cation and anion exchange resins (the volume ratio of strongly acidic cation exchange resins: resins strongly basic anion exchangers was 1: 1.5, the volume ratio of ionic resins to the crude inulin extract of flour of Helianthus tuberosus (L. 1753} L. was 1: 1.2, the duration of the resin treatment was 75 min}, to give an inulin extract from flour of Helianthus tuberosus {L. 1753) L. desalinated; (5) The inulin extract from flour of Helianthus tuberosus (L. 1753) Desalinated L. was subjected to ultrafiltration to remove small molecule impurities such as monosaccharides and oligosaccharides (membrane separation equipment: model RNF-0460, the molecular weight limit of the ultrafittration membrane was 1000 Da, with 0.2 m2 membrane area, Xiamen Filter & Me mbrane Technology Co, Ltd. ; the operating pressure differential at 1.0 MPa, with a circular flow of 2 l / min), to give the inulin extract of the flour of Helianthus tuberosus (L. 1753) L .; (6) The purified inulin solution was spray dried (the inlet temperature was 190 ° C, the outlet temperature was 90 ° C, with a feed rate of 900 ml / h), to give the inulin product from the flour of Helianthus tuberosus (L. 1753) L.
    Embodiment 4 (1) Flour of Helianthus tuberosus (L. 1753) L. produced during industrial production of inulin from Helianthus tuberosus (L. 1753) L. was ground with a food mill (manufacturer: Chengxun Industrial Co., Lid, model: CXP-DJ-S-5T); (2) At a solid-liquid ratio of 1:15 {g / m}, buffered saline (citric acid-sodium citrate buffer solution, 0.25 mol, pH 4.0-5.5) containing glycosidases (the ratio of enzymes to substrates was respectively pectinase 12 U / g, cellulase 12 U / g, hemicellulase 15 U / g, and the proportion of xylanase was 10 U / g), extraction at 48 ° C for 1 , 5h; (3) The extract was subjected to ultrafiltration to remove macromolecular impurities such as protein and pectin (membrane separation equipment: model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 20,000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co.
    Lid. ; the operating pressure differential at 1.0 MPa, with a circular flow of 2 l / min}, to give a crude inulin extract from the flour of Helianthus tuberosus (L. 1753) L .; (4) The crude inulin extract of Helianthus tuberosus (L. 1753) L. flour was desalinated and purified with cation and anion exchange resins (the volume ratio of strongly acidic cation exchange resins: resins strongly basic anion exchangers was 1: 1.2, the volume ratio of ionic resins to crude inulin extract from flour of Helianthus tuberosus (L. 1753) L. was 1: 1.4, duration treatment with resins was 88 min), to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated; (5) Inulin extract from Helianthus tuberosus (L. 1753) L. desalinated flour was subjected to ultrafiltration to remove small molecule impurities such as monosaccharides and oligosaccharides (membrane separation equipment: Model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 1000 Da, with a membrane area of 0.2 m2, purchased from Xiamen Filter & Membrane Technology Co., Ltd.; the pressure differential working at 1.0 MPa, with a circular flow of 2 min), to give the inulin extract of the flour of Helianthus tuberosus (L. 1753) L .; (6) The purified inulin solution was spray dried (the inlet temperature was 188 ° C, the outlet temperature was 92 ° C, with a feed rate of 920 ml / h), to give the inulin product from the flour of Helianthus tuberosus (L. 1758) L.
    Embodiment 5 (1) Flour of Helianthus tuberosus (L. 1753) L. produced during industrial production of inulin from Helianthus tuberosus (L. 1753) L. was ground with a food mill (manufacturer: Chengxun Industrial Co., Ltd, model: CXP-DJ-S-5T); (2) At a solid-liquid ratio of 1:25 (g / ml), buffered saline {citric acid-sodium citrate buffer solution, 0.02 mol /}, pH 4.0-5, 5) containing glycosidases (the ratio of enzymes to substrates was respectively pectinase 18 U / g, cellulase 8 U / g, hemicellulase 26 U / g, and the proportion of xylanase was 16 U / g), extraction at 52 ° C for 2.5 h: (3) The extract was subjected to ultrafiltration to remove macromolecular impurities such as proteins and pectin (membrane separation equipment: model RNF-0460, the molecular weight limit of the membrane d ultrafiltration was 20,000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co.
    Ltd. ; the operating pressure differential at 1.0 MPa, with a circular flow of 2 min), to give a crude inulin extract from the flour of Helianthus tuberosus (L. 1753) L; (4) The crude inulin extract of Helianthus tuberosus (L. 1753) L. flour was desalinated and purified with cation and anion exchange resins (the volume ratio of strongly acidic cation exchange resins: resins strongly basic anion exchangers was 1: 1.8, the volume ratio of ionic resins to crude inulin extract of flour of Helianthus tuberosus (L. 1753) L. was 1: 1.4, duration treatment with resins was 73 min), to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L. desalinated; (5) Inulin extract from Helianthus tuberosus (L. 1753) L. desalinated flour was subjected to ultrafiltration to remove small molecule impurities such as monosaccharides and oligosaccharides (membrane separation equipment: Model RNF-0460, the molecular weight limit of the ultrafiltration membrane was 1000 Da, with a membrane area of 0.2 m2, Xiamen Filter & Membrane Technology Co.
    Ltd. ; the operating pressure differential at 1.0 MPa, with a circular flow of 2 min), to give the inulin extract of the flour of Helianthus tuberosus (L. 1753) L .; (6) The purified inulin solution was spray dried (inlet temperature was 192 ° C, outlet temperature was 91 ° C, with feed rate 690ml / h)}, to give the inulin product of flour of Helianthus tuberosus (L. 1753} L.
    Comparative Example 1 The extraction conditions for traditional hot water extraction: the extraction solution was water, the solid-liquid ratio was 1:20 (g / m), the extraction time was 2 h, and the extraction temperature was 80 ° C.
    The procedures for purifying and drying the extract were the same as in the process described in this patent.
    Embodiment 6 Helianthus tuberosus (L. 1753) L. flour inulin prepared in Embodiments 1-5 and Helianthus tuberosus flour inulin {L. 1753) L. prepared in Comparative Example 1 were identified as the objects for determining the yield of inulin, the content of total saccharides, the content of 1F-fructofuranosylnystose and of fructans of higher degrees of polymerization, the content protein, uronic acid content and fructose residue configuration and type of glycosidic bond in fructans.
    The inulin yield was calculated according to Formula (1): the inulin yield (%) = (the total saccharide content - ia reducing saccharide content) / the mass of Helianthus tuberosus (L. 1753) L. x 100% Formula (1) in which, the content of total saccharides was determined by the phenol-sulfuric acid method with fructose as a standard, the content of reducing saccharides was determined by a method to dinitrosalicylic acid (DNS) with glucose as a standard.
    Determination of total saccharide content: With fructose as the monosaccharide standard, the determination method was: Dubois M, Gilles KA, Hamilton JK, et al. Colorimetric method for determination of sugars and related substances [J]. Analytical Chemistry, 1956, 28: 350-356.
    The method for determining the content of 1F-fructofuranosylnystose and fructans of higher degrees of polymerization was: Fu Q, Liang 7, Zhang X, et al. Carbohydrate separation by hydrophilic interaction liquid chromatography on a ‘click maltose column {J]. Carbohydrate Research, 2010, 345: 2690-2697.
    The method for determining the protein content was: Sedmark JJ, Grossberg SE. At rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250 [J]. Analytical Biochemistry, 1979, 79: 544-552) The method for determining the content of uronic acids was: Blumenkrantz N, Asboe-Hansen G. New method for quantitative determination of uronic acids [J]. Analytical Biochemistry, 1973, 54: 484-489.
    The method of determining the configuration of the fructose residue and the type of glycosidic bond in fructans was: Bi H, Gao T, Liu D, et al. Structures of (1-> 6) -B-D-glucans from Bulgaria inquinans (Fries) and their immunological activities [J]. Carbohydrate Polymers. 2013, 93: 547-552.
    The yield results of inulin from flour of Helianthus tuberosus (L. 1753) L.
    are shown in Table 1. As shown in Table 1, compared to traditional hot water extraction, the yield of inulin in the process of extracting linulin from Helianthus tuberosus (L. 1753) L.
    described in this patent was increased by 24.58-39.74%. Table 1 Comparison of the yields of Vinulin from flour of Helianthus tuberosus (L. 1753) L. by traditional hot water extraction and the preparation process of this patent Extraction Mode de | Fashion of | Fashion of | Traditional Fashion Fashion | achievement | achievement | achievement | achievement | water-based realization | 1 2 3 4 5 hot Yield | 25.26 33.38 34.12 35.30 34.36 31.47 inulin (%)
    The results of the inulin composition of flour of Helianthus tuberosus (L. 1753) L. are shown in Table 2. As shown in Table 2, compared to the traditional hot water extraction, the Helianthus tuberosus (L. 1753) L. flour Pinulin purity was increased by 2.4 5.2%, the total saccharide content was increased by 3.9% -7.7%, the content of 1F-fructofuranosylnystose and fructans of higher degrees of polymerization in inulin was increased by 3.7% -6.9%. Table 2 Comparison of the compositions of inulin from flour of Helianthus tuberosus (L. 1753) L. by traditional hot water extraction and the preparation process of the Extraction Mode disclosure of | Fashion of | Fashion of | Fashion of | Traditional fashion] | realization | realization | realization | realization | realization e in water | ni n2 n3 n4 5 Content in | 90.11 94.56 93.59 97.02 96.43 96.25 total saccharides (%) 1F content- | 72.92 78.92 76.03 77.08 76.85 75.64 fructofuranosylnystose and fructans of higher degrees of polymerization in Protein content | 0.31 0.25 0.22 0.27 0.34 0.28 a EE IP Acid content | 0.21 0.48 0.46 0.52 0.55 0.61 me RE RE IE OJ The configuration of the | —1) -B-D-Fruf- (2— fructose residue and | B-D-Fruf- (2— type of bond
    AE Embodiment 7 The Helianthus tuberosus (L. 1753} L. flour inulin products prepared in Embodiments 1-5 and commercial inulin (Orafit® HIS Inulin powder, batch number: RHWCJBAC / J8 ) were identified as the objects to determine the total antioxidant capacity.
    The method for determining the total antioxidant capacity was: to a 1.5 ml EP tube were respectively added 400 μl of 5 mM ABTS solution /, 250 μl of 1.0 μM HRP solution /, 10 Hi of 6.0 mM H2O2 solution, and 340 µl of 50 MMA phosphate buffered solution pH 7.5, the total volume of the mixed solution was 1 ml, which included 2 mM PABTS), 15 µM H2O2, 0.25 µM HRP; and 10 µl of a test sample solution were then added, the test temperature was 25 ° C: vortex for 10s, the reaction solution was transferred to a cuvette, which was left to stand for min in The darkness, and the light absorbance values at 730 nm were read.
    The results of the total antioxidant capacities of the samples were expressed in Trolox equivalents (x milligrams of water soluble vitamin E in equivalent per gram of sample, TEAC (mgTE / g)). Table 3 Comparison of the Total Antioxidant Capacities of Commercial Inulin and Helianthus tuberosus (L. 1753) L. Flour Inulin by the Inulin Mode Disclosure Preparation Method! Fashion | Fashion | Fashion! Commercial mode | achievement | achievement | achievement | achievement | achievement ee ee ee ee TEAC 2.53 10.24 | 382 + 0.29 | 3.69 + 0.16 | 3.94 +0.31 3.72 + 0.23 | 3.85 + 0.33 eee [Ee e re pros joe The results are shown in Table 3. As shown in Table 3, the total antioxidant capacity (TEAC value) was increased from 45.85-55.75 % compared to commercial linulin.
    Embodiment 8 Effects of Helianthus tuberosus (L. 1753) L. flour inulin prepared in Embodiments 1-5 on improving residual gastric contents and gastrointestinal hormone levels in DF rats Establishment of rat models with functional dyspepsia (DF): 54 male SD rats were weighed and randomly grouped into 9 groups: the blank control group (C), the model group (M), the domperidone group (D), domperidone + commercial inulin (D + commercial inulin), domperidone + inulin obtained in Embodiment 1 (D + Embodiment 1), domperidone + inulin obtained in Embodiment 2 (D + Embodiment 2), domperidone + inulin obtained in Embodiment 3 (D + Embodiment 3), domperidone + inulin obtained in Embodiment 4 (D + Embodiment 4), domperidone + inulin obtained in Embodiment 5 (D + Embodiment 5), except on of the blank group, each remaining group was modeled after being slightly improved according to the documents (Guo Haijun, et al., 2000), while giving the tail pinching stimulation, a normal diet on odd days and a fast on even days, each drug was dissolved with distilled water, the volume administered was 10 ml / kg, once a day, the duration of administration was 15 days, with specific treatments indicated in the Table 4 below.
    Table 4 PTE har Ta experimental group ———— Blank group (C) No treatment, breeding | No treatment, breeding TE me TE hama Model group (M) Tail pinch + Tail pinch +
    NN Domperidone group (D) Tail pinching + | Tail pinch + AND th eme | Domperidone + inulin | Tail pinching + | Tail nipping + commercial (D + inulin Irregular feeding Irregular feeding, commercial) 3.1 mg / kg] (D), 1.5 g / kg /] {inulin) Domperidone + Inulin | Tail Pinch + Tail Pinch + Embodiment 1 (D + | Irregular Feeding Irregular Feeding, Embodiment 1) 3.1 mg / kg / d (D), 1.5 g / kg inulin) Domperidone + Inulin | Tail pinching + | Tail pinching + Embodiment 2 (D + | Irregular feeding Irregular feeding, Embodiment 2) 3.1 mg / kg] (D), 1.5 g / kg / d (inulin) Domperidone + Inulin du | Tail pinching + | Tail Pinching + Embodiment 3 (D + | Irregular Feeding Treeclock Feeding, Embodiment 3) 3.1 mg / kb / d (D), 1.5 g / kg / d (inulin) Domperidone + Inulin | Tail pinching + | Tail Pinching + Embodiment 4 (D + | Irregular Feeding Irregular Feeding, Embodiment 4) 3.1 mg / kg (D), 1.5 g / kg / d (inulin)
    Domperidone + Inulin | Tail Pinch + Tail Pinch + Embodiment 5 (D + | Irregular Feeding Irregular Feeding, Embodiment 5) 3.1 mg / kg; (D), 1.5 g / kg] (inulin) Determination of gastric emptying rate: rats in each group were fasted but had free access to water for 12 h, and each rat received 3 mi of 0.1% methyl orange solution by gavage, after 10 minutes he was anesthetized with 5% chloral hydrate, and blood was drawn from the heart: and the abdominal cavity was opened quickly to collect the entire stomach, which was incised along the greater curvature of the stomach, the stomach contents were rinsed with 5 ml of saline solution suitably in a test tube, the pH of which was adjusted to 6-6.5 with 5% sodium bicarbonate solution and measured at 10 ml, centrifuged at 3000 rpm for 10 minutes to collect the supernatant; further mixed evenly with 3 ml of 0.1% methyl orange solution and 7 ml of saline solution, centrifuged to collect the supernatant as a standard solution; the absorbance was determined at 420 nm, and the residual level of methyl orange in the stomach of the rats of each group was calculated according to the formula below: the level of methylcrange in the stomach (%) = absorbance of methyl orange in the stomach / the absorbance of methyl orange in the standard solution x 100%. (See Cao Feng et al, 2009; QIU X J =, 2011)
    Determination of MTL, Gas, Gréhline, 5-HT contents: treatment on plasma samples: ethylenediaminetetraacetate of dipotassium (EDTA-K2) was chosen as anticoagulant, the blood was mixed uniformly in a blood collection tube containing the anticoagulant, and centrifuged for 20 min at 4 ° C and 3000 rpm to collect the supernatant as the plasma to be tested.
    Processing on serum samples: after being obtained by blood sampling, the blood was allowed to stand to coagulate naturally, centrifuged for 20 min at 4 ° C and 3000 rpm to collect the supernatant as serum to test.
    The determination of the MTL, Gas, Gréhline and 5-HT contents was carried out by a DAS-ELISA process, the specific operating steps of which were carried out according to the instructions of the kit.
    The linear regression equation of the standard curve was plotted with the concentrations of the standard as horizontal coordinates and the OD values as vertical coordinates, the corresponding sample concentrations were calculated based on the OD values of the samples, and then multiplied by the dilution factors to calculate the levels.
    Table 5 Comparison of Effects of Commercial Vinulin and Pinulin from Helianthus tuberosus (L. 1753) L. Flour by the Preparation Method of the Disclosure, Each Co-administered with Domperidone, on Gastric Emptying Rate and Contents of MTL, Gréhline, Gas, 5-HT in DF rats Rate of | MTL Gréhline Gas 5-HT draining (pg / ml; (pg / ml} {pg / m]) (pg / rnl) gastric (%) Group 40.53 +3.71 | 890.58 + 32.55 | 1797 , 59 + 9322 | 482.19 + 46.54 | 1132.56 + 93.35 me EE eee ee ee Inulin 42.68 +4.55 | 903.45 + 21.42 | 1725.44 + 89.02 | 473.36 £ 39.61 | 1044.32 + 124.48 commercial + Domperidone Inulin du Mode | 46.72 £ 3.62 | 1095.83 + 35.28 | 1704.36 + 87.53 | 453.42 437.83 | 1003.29 + 103.64 of realization 1 + Dompéridone Inufine du Mode | 47.37 +3.86 | 1124.48 + 31.17 | 1693.52+ 82.30 | 437.29 + 40, 88 | 1072.24 + 110.27 of realization 2 + Domperidone Inulin du Mode | 50.53 +4.13 | 1135.61 + 30.53 | 1 683.46 + 84.29 | 446.63 + 41, 08 | 987.50 + 108.46 + Mode domperidone inulin | 47.81 + 4.07 | 1,088.46 + 29.93 | 1,699.78 188.76 | 448.21 + 38.65 | 993.04 + 112.21 + Domperidone Inulin du Mode | 48.72+ 4.11 | 1,035.37 + 36.82 | 1,702.84 +83.34 | 451.30 + 49.73 | 1,004.27 + 105, 88 of achievement 5 + Domperidone The effects of inulin from flour of Helianthus tuberosus (L. 1753) L. prepared in Embodiments 1-5 on improving gastric contents trough and gastrointestinal hormone levels in DF rats were greater: compared to inulin from Helianthus tuberosus (L. 1753) Commercial L., in the experimental group of inulin prepared in the disclosure, the gastric emptying rate was increased by 9.47-18.39%, the MTL content was increased by 14.60-25.70 %, the Grelin content was reduced by 1.22-2.43%, the Gas content was reduced by 4.21-7.62%, and the 5-HT content was reduced by 3.84-5, 46%.
    Mode of realization 9 Experimental method on the diversity of the intestinal flora
    1. Sample collection 1) Preparation of materials of cotton, alcohol, forceps, sterile cryopreservation tubes, markers, disposable PE and rubber gloves, Zip-lock bags of various sizes, a small amount of liquid nitrogen. ‘2) (D) Find as clean a bench as possible in the dissection room and wipe it down with alcohol, trying to avoid contamination by exogenous bacteria; (Z) marking of cryopreservation tubes and numbering of Zip-lock bags; (3) abdominal massage on rats (or tilting), to defecate them on the bench and conditioning in sterile cryopreservation tubes, with rapid placement in liquid nitrogen, the samples were stored in a refrigerator at -80 ° C within 2 h after collection, ready to use; (2) wiping the benchtop with alcohol, continue to sample;
    2. DNA extraction and PCR amplification Total DNA extraction was performed according to the instructions of the EZNA® soil kit (Omega Bio-tek, Norcross, GA, US), DNA concentrations and purities were were detected with NanoDrop2000, and the DNA extraction mass was detected by 1% agarose gel electrophoresis; the viable regions V3-V4 were amplified by PCR with the pair of primers of 338F-5'-actcctacgggaggcagcag-3 '(SEQ ID No.1) and 8068 5'-ggactachvgggtwtctaat-3' (SEQ ID No.2} , amplification procedures were: pre-degeneration at 95 ° C for 3 min, 27 cycles (degeneration at 95 ° C for 30 s, hybridization at 55 ° C for 30 s, extension at 72 ° C for 30 s), finally extension at 72 ° C for 10 minutes (PCR instrument: model ABI GeneAmp® 9700) The amplification system was 20 µl, 4 µl of 5 * FastPfu buffer solution, 2 µl of 2.5 mM ONTPs, 0.8 µl of primers (5 µM), 0.4 µl of FastPfu polymerases; 10 µg of DNA templates.
    3. Illumina Miseq Sequencing The PCR products were recovered using 2% agarose gel, purified by a DNA extraction kit on gei AxyPrep (Axygen Biosciences, Union City, CA, USA) , eluted with Tris-HCl, and detected using 2% agarose electrophoresis. They were detected and quantified with QuantiFluor "M-ST (Promega, USA). The purified amplified fragments were constructed and integrated into a 2 * 300 PE library according to the standard service specification of the Illumina MiSeq sequencer (IIlumina , SanDiego, United States).
    Steps in building a bank: (1) connecting a Y-shaped adapter; (2) removal of self-ligated fragments from the adapter by screening with magnetic beads; (3) PCR amplification was used for enrichment of library templates; (4) degeneration with sodium hydroxide, to produce single stranded DNA fragments. Sequencing was performed on a Miseq PE300 sequencer from lumina Corp. (Shanghai Majorbio Bio-pharm Technology Co., Ltd).
    4. Data processing The initial sequences for sequencing were monitored with Trimmomatic software, and spliced with FLASH software: (1) setting a window of 50 bp (base pairs), if the mean mass value at the inside of the window was below 20, the bases at the rear end were truncated starting with the window to eliminate sequences having lengths below 50 bp after removing the mass check; (2) the alignment with the barcode had to be precise, allowing 2 base non-alignments in the primers, and removing fuzzy bases: (3) the sequences at both ends were spliced according to the overlap overlapping bases, the overlap having to be greater than 10 bp, by eliminating the sequences which cannot be spliced: the sequences were grouped by OUT with UPARSE software (version 71 http://drive5.com/uparse/) according to 97% similarity; and the chimeras were inactivated with UCHIME software. Each sequence was classified and scored using the RDP classifier (http // rdp.cme.msu.edu /), by comparison with the Silva database (SSU123), and setting the comparison threshold to 70%.
    Table 6 Comparison of the effects of commercial inulin and Helianthus tuberosus (L. 1753) L. flour inulin prepared in Embodiments 1-5, each administered in conjunction with domperidone, on Chao indices , ACE, Shannon and Simpson on the diversity of intestinal flora in rats DF Tee AGE Te Shannon eg Sion Group 716 +21 742 + 38 0.021 + 0.011 4.9 + 0.2 me EE (Pe Inulin 652 + 27 667 + 33 0.042 = 0.012 | 3.9 + 0.2 commercial + Domperidone inulin du | 603 + 18 642 + 35 0.033 + 0.007 42 + 02 Embodiment 1 + Domperidone Inulin du | 610 +24 637 + 42 0.031 + 0.005 | 4 , 3 + 0; 1 Embodiment 2 + Domperidone inulin du | 593 + 38 632 + 30 0.027 +0.009 45102 Embodiment 3 + Domperidone Inulin du | 599 + 31 646 + 28 0.029 + 0.009 | 44x02 Embodiment 4+ Domperidone inulin du | 607 + 26 639 + 39 0.030 + 0.008 | 42 + 0; 1 Embodiment 5 + Domperidone Results were as shown in Table 6, co inulin mmercial and the inulin obtained in Embodiments 1-5 could all reduce the Chao and ACE indices of gut flora richness, reduce the Shannon index of flora uniformity, increase the Simpson index of flora uniformity. uniformity of flora in DF rats.
    Compared to commercial inulin, the Helianthus tuberosus (L. 1753) L. flour inulin prepared in Embodiments 1-5 exhibited greater abilities to alleviate disturbances of the intestinal flora in DF rats.
    The foregoing were only the preferable detailed descriptions of the present invention. To be noted by those skilled in the art that several improvements and modifications could be made without departing from the principle of the present invention, which should also be considered to be. the scope of the present invention.
    权利要求:
    Claims (10)
    [1]
    1. Process for the preparation of Helianthus tuberosus flour inulin {L. 1753} L. to improve functional dyspepsia, which is characterized in that, which includes the following steps: 1) mixing of Helianthus tuberosus (L. 1753) L. flour powders with a buffer solution containing glycosidases, and extraction enzymatic, to give an extract; 2) ultrafiltration of the extract to remove macromolecular impurities such as proteins and pectin, to give a crude inulin extract of Helianthus tuberosus (L. 1753) L flour; 3) desalination and purification of the crude inulin extract from the flour of Helianthus tuberosus (L. 1753) L., to give an inulin extract from the flour of Helianthus tuberosus (L. 1753) L., desalinated; 4) ultrafiltration of inulin extract from the flour of Helianthus tuberosus (L. 1753) L.
    desalinated a second time to remove small molecule impurities such as monosaccharides and oligosaccharides, drying, to give Helianthus tuberosus (L. 1753) L. flour inulin.
    [2]
    2. Preparation process according to claim 1, which is characterized in that the buffer solution containing glycosidases in step 1) comprises, with a buffer solution of citric acid-sodium citrate at 0.01-0, 5 mol as solvent, the following glycosidases: pectinase, cellulase, hemicellulase and xylanase; the ratio of the mass of the flour powders of Helianthus tuberosus (L. 1753) L. to the activities of pectinase, cellulase, hemicellulase and xylanase is, in order, 1 g: 3-20U : 5-15 U: 10-30 U: 5-20 U.
    [3]
    3. Preparation process according to claim 1 or 2, which is characterized in that the ratio of the mass of the flour powders of Helianthus tuberosus (L. 1753) L. to the volume of the buffer solution containing glycosidases in the step 1) is 1 g: 10-30 mi; The temperature of the enzymatic extraction is 45-55 ° C, the time of the enzymatic extraction is 1-3 h.
    [4]
    4. Preparation process according to claim 1, which is characterized in that the limiting molecular weight of the ultrafiltration membrane used in the ultrafiltration of step 2) is 20,000 Da, with a membrane area of 0, 2 m2; the pressure differential in the ultrafiltration operation is 1.0 MPa, with a circular flow of 2 min.
    [5]
    5. Preparation process according to claim 1, which is characterized in that the desalination and purification in step 3} are carried out with cation and anion exchange resins; the volume ratio of strongly acidic cation exchange resins: strongly basic anion exchange resins is 1: 1-2: the ratio of the total volume of ionic resins to the volume of the crude inulin extract of flour of Helianthus tuberosus ( L. 1753) L. is 1: 1—1.5: the duration of desalination and purification is 60-90 min.
    [6]
    6. Preparation process according to claim 1, which is characterized in that the molecular weight limit of the ultrafiltration membrane used in the ultrafiltration for the second time in step 3) is 1000 Da, with a membrane area of 0.2 m2, an operating pressure differential of 1.0 MPa, and a circular flow of 2 l / min; the drying includes spray drying; the inlet temperature of spray drying is 185-195 ° C, the outlet temperature is 85-95 ° C, with a feed rate of 800-1000 ml / h.
    [7]
    7. Flour inulin of Helianthus tuberosus (L. 1753) L. for improving functional dyspepsia prepared by the preparation process according to any one of claims 1 to 6, which is characterized in that, the purity of flour inulin of Helianthus tuberosus (L. 1753) L. is = 91%: the content of total saccharides is 293%, in which the content of 1F-fructofuranosylnystose and of fructans of higher degrees of polymerization is 275%; the uronic acid content is <0.34%: and the protein content is <0.61%.
    [8]
    8. Helianthus tuberosus (L. 1753) L. flour inulin according to claim 7, which is characterized in that the total antioxidant capacity of [Helianthus tuberosus (L. 1753) L. flour inulin is expressed by the TEAC; TEAC is x milligrams of water soluble vitamin E in equivalent per gram of inulin sample from Helianthus tuberosus (L. 1758) L flour; the total antioxidant capacity of inulin from flour of Helianthus tuberosus (L. 1753) L. is 3.56-4.25 mg TE / g.
    [9]
    9. Use of Helianthus tuberosus (L. 1753) L. flour inulin according to claim 7 or 8 in the preparation of medicaments for the treatment of functional dyspepsia.
    [10]
    10. Use of the finulin from flour of Helianthus tuberosus (L. 1753) L. according to claim 7 or 8 in health care products for improving the residual level of gastric contents and the levels of gastrointestinal hormones, disorders. of the intestinal flora and / or the level of oxidative stress.
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    同族专利:
    公开号 | 公开日
    CN110903413A|2020-03-24|
    BE1027863A1|2021-07-09|
    CN110903413B|2021-09-07|
    引用文献:
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    CN1919855A|2006-09-21|2007-02-28|贵州大学|Method of extracting inulin assisted with microwave technique|
    US20100119651A1|2008-11-12|2010-05-13|Fan Shufeng|Bio-Enzyme-Assisted Extraction Method for Inulin|
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    CN104292367A|2014-10-15|2015-01-21|重庆骄王天然产物股份有限公司|Refining method of inulin|CN112300300A|2020-11-02|2021-02-02|陕西森弗天然制品有限公司|Inulin purification method|
    法律状态:
    2021-08-11| FG| Patent granted|Effective date: 20210714 |
    优先权:
    申请号 | 申请日 | 专利标题
    CN201911265145.3A|CN110903413B|2019-12-11|2019-12-11|Jerusalem artichoke pulp inulin for improving functional dyspepsia and preparation method and application thereof|
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