A method for preparing Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, trifructan, and inulbiose.

Abstract

This invention discloses a method for preparing a Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, difructose triterpenoids, and inulbiose. The method includes: pulverizing dried Jerusalem artichokes to obtain Jerusalem artichoke powder; mixing the dried Jerusalem artichoke powder with water and heating to obtain a Jerusalem artichoke powder solution; cooling the Jerusalem artichoke powder solution, adding difructose triterpenoid hydrolase for enzymatic hydrolysis, inactivating the enzyme, centrifuging to obtain the extract, and obtaining a Jerusalem artichoke powder reaction solution rich in inulin, fructooligosaccharides, difructose triterpenoids, and inulbiose; and spray-drying the Jerusalem artichoke powder reaction solution to obtain the Jerusalem artichoke solid beverage. The Jerusalem artichoke solid beverage prepared by this invention has diverse functional elements, including not only the naturally present inulin but also the generated fructooligosaccharides, difructose triterpenoids, and inulbiose. Inulin, fructooligosaccharides, and difructose triterpenoids can promote mineral absorption, play a role in preventing tooth decay, promoting the growth of probiotics, and improving human immunity.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, specifically relating to a method for preparing Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, type trifructosan and inulbiose. Background Technology

[0002] Jerusalem artichoke (Helianthus tuberosus L.) is a herbaceous plant, also known as sunchoke, Jerusalem artichoke, or five-star grass. It typically flowers and fruits in autumn. The flower resembles a chrysanthemum, and the tuber contains a high content of inulin. Jerusalem artichoke tubers are a focus of research, not only because they are edible, but also because they can be used as a raw material for alcohol production. The stems, leaves, and tubers of Jerusalem artichoke possess various biological activities, including boosting immunity, anti-tumor activity, antioxidant properties, and blood sugar lowering.

[0003] Inulin is a functional dietary fiber that promotes the growth of beneficial gut bacteria, improves the gut microenvironment, regulates blood sugar levels, promotes mineral absorption, relieves constipation, and reduces the risk of cancer. Inulin can be converted into fructooligosaccharides and the novel functional sweetener difructose triterpenoids by difructose triterpenoids hydrolase. Furthermore, difructose triterpenoids can be hydrolyzed into inulbiose by difructose triterpenoids hydrolase.

[0004] Fructose oligosaccharides are a natural active substance with a sweetness 0.3-0.6 times that of sucrose. They retain the pure sweetness of sucrose but are more refreshing. This novel sweetener possesses health benefits such as regulating intestinal flora, promoting bifidobacteria growth, enhancing calcium absorption, regulating blood lipids, and preventing tooth decay. It is hailed as the most promising next-generation additive—a growth promoter—since the era of antibiotics. Type III dicfructan also has a range of physiological functions, including promoting mineral absorption, preventing tooth decay, and promoting the growth of probiotics. As a metabolite of type III dicfructan, inulinbiose theoretically has prebiotic effects. It is easily soluble in water and tastes sweeter than sucrose, indicating its potential as a functional sweetener.

[0005] Currently, there are few products developed specifically for Jerusalem artichoke in China. The market only offers primary agricultural products such as dried Jerusalem artichoke and insulin tea, lacking in-depth processing of these high-inulin-content products. Therefore, increasing the added value and nutritional value of Jerusalem artichoke is a technical problem that needs to be solved. Summary of the Invention

[0006] The purpose of this section is to outline some aspects of embodiments of the present invention and to briefly describe some preferred embodiments. Simplifications or omissions may be made in this section, as well as in the abstract and title of this application, to avoid obscuring the purpose of these documents; however, such simplifications or omissions should not be construed as limiting the scope of the invention.

[0007] In view of the problems existing in the above and / or prior art, the present invention is proposed.

[0008] Therefore, the purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, type trifructosine and inulbiose.

[0009] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a method for preparing Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, trifructosan, and inulbiose, comprising,

[0010] The dried Jerusalem artichokes were pulverized to obtain dried Jerusalem artichoke powder.

[0011] Jerusalem artichoke powder was mixed with water and heated to obtain a Jerusalem artichoke powder solution.

[0012] The Jerusalem artichoke powder solution was cooled, and type III fructosine hydrolase was added for enzymatic hydrolysis. After enzyme inactivation, the extract was collected by centrifugation to obtain a Jerusalem artichoke powder reaction solution rich in inulin, fructooligosaccharides, type III fructosine and inulbiose.

[0013] The Jerusalem artichoke solid beverage was prepared by spray drying the Jerusalem artichoke powder reaction solution.

[0014] As a preferred embodiment of the preparation method described in this invention, the dried Jerusalem artichoke includes naturally air-dried, clean and dirt-free dried Jerusalem artichoke.

[0015] In a preferred embodiment of the preparation method described in this invention, the pulverization is performed to a 100-mesh sieve.

[0016] As a preferred embodiment of the preparation method of the present invention, the step of mixing Jerusalem artichoke powder with water is wherein the mass ratio of Jerusalem artichoke powder to water is 1:10 to 30.

[0017] In a preferred embodiment of the preparation method described in this invention, the Jerusalem artichoke powder solution is prepared by heating, wherein the heating temperature is 100°C and the heating time is 10 min.

[0018] In a preferred embodiment of the preparation method described in this invention, the Jerusalem artichoke powder solution is cooled to 40°C.

[0019] As a preferred embodiment of the preparation method described in this invention, the type III difructosyl anhydride hydrolase is derived from Bifidobacterium cebidarum, and its nucleotide sequence is SEQ ID NO.1.

[0020] As a preferred embodiment of the preparation method described in this invention, the content of type III difructose anhydride hydrolase is 0.05‰ of the mass of Jerusalem artichoke powder, the hydrolysis time is 2-6 h, the pH is 5.5, and the specific enzyme activity of type III difructose anhydride hydrolase is 40.18 U / mg.

[0021] As a preferred embodiment of the preparation method described in this invention, the spray drying process has the following parameters: inlet air temperature 180°C, outlet air temperature 120°C, and compressed air pressure in the pipeline 3 bar.

[0022] Another objective of this invention is to overcome the shortcomings of the prior art and provide a Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, trifructosan, and inulbiose.

[0023] Beneficial effects of this invention:

[0024] (1) This invention uses primary agricultural product dried Jerusalem artichoke. By processing the dried Jerusalem artichoke, the added value of the product is increased, and the advantage of Jerusalem artichoke's high inulin content is fully utilized.

[0025] (2) The Jerusalem artichoke solid beverage prepared by the present invention has a variety of functional elements, including not only the inulin contained therein, but also the oligofructose (GF2, GF3, GF4), type tri-dispermanent and inulbiose; among them, inulin, oligofructose and type tri-dispermanent can promote the absorption of minerals, play a role in anti-caries, proliferate probiotics and improve human immunity.

[0026] (3) The type III difructosyl anhydride hydrolase used in the product of this invention can not only generate oligofructose and type III difructosyl anhydride using inulin as a substrate, but also hydrolyze type III difructosyl anhydride into potential prebiotic inulinobiose. Therefore, in actual production, the type III difructosyl anhydride hydrolase used in this product can exert the effects of both type III inulin glycosyltransferase and type III difructosyl anhydride hydrolase, reducing the cost of using two enzymes to prepare related products. Attached Figure Description

[0027] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort. Wherein:

[0028] Figure 1 This is the HPLC chromatogram of inulin, the standard of this invention;

[0029] Figure 2 The HPLC chromatograms of the standards GF2, GF3, and GF4 of this invention are shown below.

[0030] Figure 3 This is an HPLC chromatogram of the substrate difructosan and the product inulinobiose when the enzyme preparation is catalyzed by difructosan tris as a substrate in the present invention.

[0031] Figure 4This is an HPLC chromatogram of the components of the Jerusalem artichoke powder solution before reaction in Example 3 of the present invention;

[0032] Figure 5 This is an HPLC chromatogram of the components of Jerusalem artichoke powder solution after reaction in Example 3 of the present invention. Detailed Implementation

[0033] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0034] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0035] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that is mutually exclusive with other embodiments.

[0036] Example 1

[0037] This embodiment provides a method for preparing an enzyme preparation with type III difructose anhydride hydrolase activity, the main steps of which are as follows:

[0038] (1) Plasmid construction: The gene of type III difructosyl anhydrase from Bifidobacterium cebidarum (nucleotide sequence as shown in SEQ ID NO.1) was constructed into the multiple cloning site of the pET-22b(+) plasmid using gene synthesis methods to obtain the recombinant plasmid pET-22b(+)-BcebDFAase-Ⅲ.

[0039] Specifically, the amino acid sequence of type III difructose anhydride hydrolase from D. gerundensis A4 (GenBank accession number: QTO55438.1) was compared with the sequence in the non-redundant protein sequence database on NCBI using the BLAST (basic local alignment tool). The comparison results revealed a potential type III difructose anhydride hydrolase gene (SEQ ID NO.1) from Bifidobacteriumcebidarum.

[0040] The 5′ and 3′ ends of the nucleotide sequence corresponding to the coding gene were introduced into the Nde I and Xho I restriction sites, respectively. At the same time, a 6-histidine tag was added before the stop codon for the separation and purification of the protein by nickel ion affinity chromatography. Subsequently, the synthesized gene was ligated into the vector pET-22b(+) to obtain the recombinant plasmid, in which pET-22b(+) contains the ampicillin resistance gene for the screening of recombinants.

[0041] (2) Transformation and induced expression of recombinant plasmids: The recombinant plasmid pET-22b(+)-BcebDFAase-Ⅲ constructed in step (1) was transformed into E. coli BL21(DE3) competent cells, plated on LB plates and incubated upside down overnight. Single colony clones were picked and cultured in LB liquid medium at 37°C and 200 rpm overnight. The cells were then inoculated at an inoculum of 2% (v / v) into LB liquid medium and cultured at 37°C until OD600. 600 The value was 0.6, IPTG was added to a final concentration of 0.5 mM, and the mixture was induced at 28℃ and 200 rpm for 6 h. The fermentation broth was then collected.

[0042] (3) Isolation and purification of type III difructose anhydride hydrolase: The fermentation broth was centrifuged at 4℃ and 10000rpm for 30min to obtain the cell pellet. 20mL of buffer (50mM PBS, 500mM NaCl, pH adjusted to 5.5) was added to the cell pellet to fully resuspend the cells. The cells were then subjected to ultrasonic disruption in an ultrasonic cell disruptor to obtain crude enzyme solution, which was filtered through a 0.22μm microporous membrane for later use.

[0043] Protein purification was performed using a nickel ion affinity chromatography column to obtain the target protein. The purified type III difructosyl anhydrase reached electrophoretic purity.

[0044] Because histidine was added during gene construction, enzyme purification was primarily performed using nickel affinity chromatography. Deionized water (approximately 6–12 column volumes) was pumped into the nickel affinity chromatography column at 4°C using a constant flow pump, followed by 5 column volumes of low-salt binding buffer (500 mmol / L). -1 NaCl, 50 mmol / L -1 Equilibrate the column with acetate buffer (pH 5.5), and load the supernatant from the membrane onto the column; wash with a buffer containing a low concentration of imidazole (500 mmol / L). -1 NaCl, 50 mmol / L -1 Imidazole, 50 mmol / L -1 Elute off contaminating proteins with acetate buffer (pH 5.5), followed by elution buffer (500 mmol / L) containing a high concentration of imidazole. -1NaCl, 500 mmol / L -1 Imidazole, 50 mmol / L -1 The target protein was eluted with acetate buffer (pH 5.5); the collected enzyme was dialyzed overnight with EDTA-containing dialysate to remove high concentrations of imidazole and salts, and then residual EDTA was removed with EDTA-free dialysate to obtain pure enzyme, which was stored at 4°C.

[0045] (4) Enzyme activity assay of type III difructan hydrolase: 1 mL of reaction system contains 1% (w / v) type III difructan and 50 mmol / L -1 Acetate buffer (pH 5.5) and 8 nmol L -1 The enzyme solution was reacted at 40℃ for 10 min, followed by enzyme inactivation in a boiling water bath for 10 min. After the boiling water bath, the reaction solution was then heated at 10000 rpm. -1 Centrifuge for 5 min under the specified conditions. Filter the supernatant through a 0.22 μm microporous membrane, determine the amount of inulinobiose produced using the DNS method, and calculate the enzyme activity of type III difructosylhydrolase.

[0046] Unit enzyme activity is defined as the amount of enzyme required to generate 1 μmol of inulinbiose per minute under optimal reaction conditions (pH 5.5, 40℃).

[0047] The type III difructosylhydrolase prepared in Example 1 has an enzyme activity of 40 U / mg, an optimal temperature of 40℃, and an optimal pH of 5.5.

[0048] The HPLC chromatogram of the reaction between the substrate difructan and the product inulinbiose when the enzyme preparation is used as a substrate for catalysis is shown below. Figure 3 The reaction conditions were 40℃, optimal pH 5.5, and 10 min. The substrate was type tri fructosine standard, and the enzyme dosage was 0.03‰. This indicates that the enzyme can exert its hydrolytic activity, hydrolyzing type tri fructosine into the potential prebiotic inulinbiose.

[0049] Example 2

[0050] This embodiment provides a method for preparing Xuzhou white Jerusalem artichoke solid beverage (2h), the main steps of which are:

[0051] (1) Preparation of Jerusalem artichoke powder: Select white Jerusalem artichoke slices from Xuzhou that are naturally air-dried without additives, mold, insects, or pollution, and grind them into powder using a multi-functional pulverizer and then pass them through a 100-mesh sieve for later use.

[0052] (2) Preparation of Jerusalem artichoke powder solution: Weigh 6g of Jerusalem artichoke powder and dissolve it in purified water at a weight ratio of 1:15. Inactivate the enzyme at 100℃ for 10min, which also serves to dissolve the inulin by heating.

[0053] (3) Preparation of Jerusalem artichoke reaction solution: Cool the Jerusalem artichoke powder solution to 40°C, add type III difructose anhydride hydrolase preparation (enzyme mass is 0.05‰ of Jerusalem artichoke powder mass) and hydrolyze for 2 hours. After the hydrolysis is completed, inactivate the enzyme at 100°C for 10 minutes and then centrifuge to collect the extract.

[0054] (4) Preparation of Jerusalem artichoke solid beverage: The Jerusalem artichoke powder extract was spray-dried at an inlet air temperature of 180°C and an outlet air temperature of 120°C. The compressed air pressure in the pipeline was 3 bar. The product was then packaged into solid beverage packets using packaging equipment, with each packet containing 6 ± 0.5 g.

[0055] Example 3

[0056] This embodiment provides a method for preparing Xuzhou white Jerusalem artichoke solid beverage (6h), the main steps of which are as follows:

[0057] (1) Preparation of Jerusalem artichoke powder: Select white Jerusalem artichoke slices from Xuzhou that are naturally air-dried without additives, mold, insects, or pollution, and grind them into powder using a multi-functional pulverizer and then pass them through a 100-mesh sieve for later use.

[0058] (2) Preparation of Jerusalem artichoke powder solution: Weigh 6g of Jerusalem artichoke powder and dissolve it in purified water at a weight ratio of 1:15. Inactivate the enzyme at 100℃ for 10min, which also serves to dissolve the inulin by heating.

[0059] (3) Preparation of Jerusalem artichoke reaction solution: Cool the Jerusalem artichoke powder solution to 40°C, add type III difructose anhydride hydrolase preparation (enzyme mass is 0.05‰ of Jerusalem artichoke powder mass) and hydrolyze for 6 hours. After the hydrolysis is completed, inactivate the enzyme at 100°C for 10 minutes and then centrifuge to collect the extract.

[0060] (4) Preparation of Jerusalem artichoke solid beverage: The Jerusalem artichoke powder extract was spray-dried at an inlet air temperature of 180°C and an outlet air temperature of 120°C. The compressed air pressure in the pipeline was 3 bar. The product was then packaged into solid beverage packets using packaging equipment, with each packet containing 6 ± 0.5 g.

[0061] Example 3: HPLC chromatograms of the Jerusalem artichoke powder solution before and after the reaction are shown in the figure. Figure 4 , Figure 5 In the chromatogram, the peak that has the same retention time as the standard in the sugar column is the peak of the product, which can be used for both product identification and content detection.

[0062] Example 4

[0063] This embodiment provides a method for preparing a Shandong red Jerusalem artichoke solid beverage (2h), the main steps of which are as follows:

[0064] (1) Preparation of Jerusalem artichoke powder: Select naturally air-dried red Jerusalem artichoke slices from Shandong Province that are free from additives, mold, insects and pollution, and grind them into powder using a multi-functional pulverizer and then pass them through a 100-mesh sieve for later use.

[0065] (2) Preparation of Jerusalem artichoke powder solution: Weigh 6g of Jerusalem artichoke powder and dissolve it in purified water at a weight ratio of 1:20. Inactivate the enzyme at 100℃ for 10min, which also serves to dissolve the inulin by heating.

[0066] (3) Preparation of Jerusalem artichoke reaction solution: Cool the Jerusalem artichoke powder solution to 40°C, add type III difructose anhydride hydrolase preparation (enzyme mass is 0.05‰ of Jerusalem artichoke powder mass) and hydrolyze for 2 hours. After the hydrolysis is completed, inactivate the enzyme at 100°C for 10 minutes and then centrifuge to collect the extract.

[0067] (4) Preparation of Jerusalem artichoke solid beverage: The Jerusalem artichoke powder extract was spray-dried at an inlet air temperature of 180°C and an outlet air temperature of 120°C. The compressed air pressure in the pipeline was 3 bar. The product was then packaged into solid beverage packets using packaging equipment, with each packet containing 6 ± 0.5 g.

[0068] Example 5

[0069] This embodiment provides a method for preparing a Shandong red Jerusalem artichoke solid beverage (6h), the main steps of which are as follows:

[0070] (1) Preparation of Jerusalem artichoke powder: Select naturally air-dried red Jerusalem artichoke slices from Shandong Province that are free from additives, mold, insects and pollution, and grind them into powder using a multi-functional pulverizer and then pass them through a 100-mesh sieve for later use.

[0071] (2) Preparation of Jerusalem artichoke powder solution: Weigh 6g of Jerusalem artichoke powder and dissolve it in purified water at a weight ratio of 1:25. Inactivate the enzyme at 100℃ for 10min, which also serves to dissolve the inulin by heating.

[0072] (3) Preparation of Jerusalem artichoke reaction solution: Cool the Jerusalem artichoke powder solution to 40°C, add type III difructose anhydride hydrolase preparation (enzyme mass is 0.05‰ of Jerusalem artichoke powder mass) and hydrolyze for 6 hours. After the hydrolysis is completed, inactivate the enzyme at 100°C for 10 minutes and then centrifuge to collect the extract.

[0073] (4) Preparation of Jerusalem artichoke solid beverage: The Jerusalem artichoke powder extract was spray-dried at an inlet air temperature of 180°C and an outlet air temperature of 120°C. The compressed air pressure in the pipeline was 3 bar. The product was then packaged into solid beverage packets using packaging equipment, with each packet containing 6 ± 0.5 g.

[0074] The method for measuring the parameter index of this invention:

[0075] 1. Experimental Methods

[0076] The Jerusalem artichoke reaction extract after centrifugation was filtered through a 0.22 μm microporous membrane for later use. Inulin, fructooligosaccharides, and difructosan type III were determined by high performance liquid chromatography, and inulbiose was determined by a combination of high performance liquid chromatography and DNS method.

[0077] Inulin and fructooligosaccharides were identified and their content determined using a Sugar-Pak I liquid chromatography column. The mobile phase was 0.1 mM EDTA-Ca, and the flow rate was 0.4 ml / min. -1 The detector and column temperature were both 60℃, the sample loading volume was 10μl, and the detector was a Shodex differential refractive index detector.

[0078] Type III difructosan and inulinobiose were identified and their content determined using an Asahipak NH2P-504E amino column. The mobile phase was 70% acetonitrile, and the flow rate was 1 ml / min. -1 The detector and column temperature were both 30℃, the sample loading volume was 10μl, and the detector was a Shodex differential refractive index detector.

[0079] See the HPLC chromatogram of inulin standard. Figure 1 The inulin in the product was identified and its content was detected using a Sugar-PakI liquid chromatography column. The inulin standard is shown here. The peak with the same retention time in the sugar column is the inulin peak, which can also be used for subsequent content calculation.

[0080] HPLC chromatograms of standards GF2, GF3, and GF4 are shown below. Figure 2 The oligofructose in the product was identified and its content was detected using a Sugar-Pak I liquid chromatography column. The standard for oligofructose is the peak whose retention time in the sugar column is consistent with that point. This peak can also be used for subsequent content calculation.

[0081] 2. Experimental Results

[0082] As shown in Table 1, in the Jerusalem artichoke reaction solutions obtained in Examples 2 and 3, the contents of inulin and GF4 gradually decreased with the extension of reaction time.

[0083] In the 2-hour reaction system, the contents of GF2 and GF3 increased compared to before the reaction, but after 6 hours of reaction, the content of GF3 decreased compared to the 2-hour system.

[0084] The content of type III fructosine and inulin increases with the extension of reaction time. These sugars can work synergistically to improve glucose and lipid metabolism, promote mineral absorption, promote the absorption of flavonoids, prevent tooth decay, regulate the balance of intestinal flora, and enhance human immunity.

[0085] Table 3. Content of various sugars in the Jerusalem artichoke reaction solutions obtained in Examples 2 and 3

[0086]

[0087] Comparative Example 1

[0088] This comparative example uses type III inulin glycosyltransferase (derived from Arthrobacter erramosus, nucleotide sequence SEQ ID NO.2, same as the inulin glycosyltransferase in CN115868563 B) to prepare Xuzhou white Jerusalem artichoke solid beverage. The specific steps are as follows:

[0089] (1) Preparation of Jerusalem artichoke powder: Select white Jerusalem artichoke slices from Xuzhou that are naturally air-dried without additives, mold, insects, or pollution, and grind them into powder using a multi-functional pulverizer and then pass them through a 100-mesh sieve for later use.

[0090] (2) Preparation of Jerusalem artichoke powder solution: Weigh 6g of Jerusalem artichoke powder and dissolve it in purified water at a weight ratio of 1:15. Inactivate the enzyme at 100℃ for 10min, which also serves to dissolve the inulin by heating.

[0091] (3) Preparation of Jerusalem artichoke reaction solution: The Jerusalem artichoke powder solution was cooled to 55℃, and type III inulin glycosyltransferase preparation (enzyme mass was 0.05‰ of Jerusalem artichoke powder mass) was added for enzymatic hydrolysis for 6 hours. After enzymatic hydrolysis, the enzyme was inactivated at 100℃ for 10 minutes, and then the extract was collected by centrifugation. The specific activity of inulin glycosyltransferase was 311.93 U / mg, the optimal temperature was 55℃, and the optimal pH was 5.5.

[0092] (4) Preparation of Jerusalem artichoke solid beverage: The Jerusalem artichoke powder extract was spray-dried at an inlet air temperature of 180°C and an outlet air temperature of 120°C. The compressed air pressure in the pipeline was 3 bar. The product was then packaged into solid beverage packets using packaging equipment, with each packet containing 6 ± 0.5 g.

[0093] Table 4. Content of various sugars in the Jerusalem artichoke reaction solution obtained in Example 3 and Comparative Example 1.

[0094]

[0095] As can be seen from Table 4, under the same process conditions, the content of type III difructose anhydride in the Jerusalem artichoke reaction solution obtained in Example 3 increased significantly, and 9.6g of inulbiose was also produced. This indicates that the enzyme used in this patent has a stronger ability to hydrolyze inulin than the enzyme used in the comparative example, and can play the role of type III difructose anhydride hydrolase to generate potential prebiotics and sweetener inulbiose. This not only saves the cost of using two enzyme preparations, but also achieves diversification of functional components in the product.

[0096] In this invention, the type III difructosyl anhydride hydrolase is derived from probiotics. This enzyme can not only act as an inulin glycosyltransferase to catalyze inulin, but also hydrolyze type III difructosyl anhydride into the potential prebiotic inulinbiose, saving the cost of using two enzyme preparations. At the same time, the functional components in the product are diversified.

[0097] In summary, the type III difructosyl anhydride hydrolase used in the product of this invention can not only generate oligofructose and type III difructosyl anhydride using inulin as a substrate, but also hydrolyze type III difructosyl anhydride into the potential prebiotic inulinbiose. Therefore, in actual production, the type III difructosyl anhydride hydrolase used in this product can exert the effects of both type III inulin glycosyltransferase and type III difructosyl anhydride hydrolase, reducing the cost of preparing related products using two enzymes.

[0098] The nucleotide sequence listing in this invention is as follows:

[0099] SEQ ID NO.1:

[0100]

[0101] SEQ ID NO.2:

[0102]

[0103] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the present invention.

Claims

1. A method for preparing a Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, trifructosan, and inulbiose, characterized in that: include, The dried Jerusalem artichokes were pulverized to obtain dried Jerusalem artichoke powder. Jerusalem artichoke powder was mixed with water and heated to obtain a Jerusalem artichoke powder solution. The Jerusalem artichoke powder solution was cooled to 40°C, and type III difructose anhydride hydrolase was added for enzymatic hydrolysis. After enzyme inactivation, the extract was collected by centrifugation to obtain a Jerusalem artichoke powder reaction solution rich in inulin, fructooligosaccharides, type III difructose anhydride, and inulbiose. The type III difructose anhydride hydrolase was derived from... Bifidobacterium cebidarum The nucleotide sequence is SEQ ID NO.1, the mass of type III difructose anhydride hydrolase is 0.05‰ of the mass of Jerusalem artichoke powder, the hydrolysis time is 2~6 h, the pH is 5.5, and the specific enzyme activity of type III difructose anhydride hydrolase is 40.18 U / mg; The Jerusalem artichoke solid beverage was prepared by spray drying the Jerusalem artichoke powder reaction solution.

2. The preparation method according to claim 1, characterized in that: The dried Jerusalem artichokes include naturally air-dried, clean and free of dirt.

3. The preparation method according to claim 1 or 2, characterized in that: The pulverization refers to pulverizing to a 100-mesh sieve.

4. The preparation method according to claim 1, characterized in that: The method involves mixing dried Jerusalem artichoke powder with water, wherein the mass ratio of Jerusalem artichoke powder to water is 1:10~30.

5. The preparation method according to claim 1 or 4, characterized in that: The Jerusalem artichoke powder solution is prepared by heating, wherein the heating temperature is 100℃ and the time is 10 min.

6. The preparation method according to claim 1, characterized in that: The spray drying parameters are: inlet air temperature 180℃, outlet air temperature 120℃, and compressed air pressure in the pipeline 3 bar.

7. A Jerusalem artichoke solid beverage rich in inulin, fructooligosaccharides, trifructosan and inulbiose, prepared by any of the preparation methods described in claims 1, 2, 4 or 6.

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