Desert cistanche primary pulp based on Maillard reaction and preparation method thereof

Abstract

The invention discloses desert cistanche primary pulp based on Maillard reaction and a preparation method thereof, and the method comprises the following steps: adding water into cistanche and pulping to obtain cistanche primary pulp which is not debittered; adding compound protease into the cistanche primary pulp which is not debittered to carry out enzymolysis, so as to obtain cistanche enzymatic hydrolysate; adjusting the pH value of the cistanche enzymatic hydrolysate, heating and stirring for Maillard reaction, and cooling to obtain the debittered cistanche primary pulp. Through the composite enzymolysis technology, the bitter taste of bitter peptides in the cistanche pulp is effectively reduced, and possible bitter amino acids are removed through the subsequent Maillard reaction, so that the bitter taste of the finally prepared cistanche primary pulp is reduced, and the palatability is improved.

Description

Technical Field

[0001] The invention relates to the technical field of food processing, and in particular to a Cistanche deserticola puree based on the Maillard reaction and a preparation method thereof. Background Art

[0002] As a traditional Chinese medicine, Cistanche deserticola was first recorded in Shennong's Herbal Classic and is mainly distributed in the Inner Mongolia Autonomous Region, Xinjiang Uygur Autonomous Region, Ningxia Hui Autonomous Region and other northwestern desert areas. Cistanche deserticola contains a variety of bioactive ingredients such as phenylethanol glycosides, iridoid ether terpenes and their glycosides, lignans and their glycosides, sugars, etc. It has good kidney-tonifying and aphrodisiac, liver-protecting, anti-fatigue, laxative and other functions, and is known as the "desert ginseng". In 2023, desert Cistanche deserticola was officially included in the catalog of medicinal and edible homology, which means that Cistanche deserticola has the "dual identity" of food and Chinese medicinal materials. Enterprises can legally use it in food production, sales, and use, further opening up the market.

[0003] Since desert Cistanche contains more bitter amino acids and there is 5.22g of protein per 100g of Cistanche water extract, protein hydrolysis may lead to the production of bitter peptides during the development of Cistanche products. This makes the products developed with desert Cistanche as raw material bitter and poor in palatability. However, the products based on desert Cistanche in the prior art and the literature reports do not specifically treat some bitter substances in it, which makes Cistanche products uncompetitive in the market. Summary of the invention

[0004] To solve the above problems, the present invention provides a desert Cistanche puree based on the Maillard reaction and a preparation method thereof. The bitterness of bitter peptides in the Cistanche puree is effectively reduced through composite enzymatic hydrolysis technology, and possible bitter amino acids are removed through subsequent Maillard reaction, so that the bitterness of the finally prepared Cistanche puree is reduced and the palatability is improved.

[0005] To achieve the above object, the present invention proposes a method for preparing Cistanche deserticola puree based on the Maillard reaction, which comprises the following steps:

[0006] (1) adding water to the Cistanche deserticola to make a pulp, thereby obtaining a Cistanche deserticola pulp that has not been debittered;

[0007] (2) adding a composite protease to the undebittered Cistanche deserticola puree for enzymolysis to obtain a Cistanche deserticola enzymolysis solution;

[0008] (3) adjusting the pH value of the Cistanche deserticola enzymatic hydrolyzate, heating and stirring to perform Maillard reaction, and cooling to obtain debittered Cistanche deserticola puree.

[0009] According to a method for preparing a desert Cistanche puree based on the Maillard reaction of an embodiment of the present invention, the method effectively reduces the bitterness of bitter peptides in the Cistanche pulp by a composite enzymatic hydrolysis technology, and retains its high biological activity, and then removes the possible bitter amino acids by a subsequent Maillard reaction, so that the bitterness of the Cistanche puree finally prepared is reduced. Compared with the traditional Cistanche puree, the de-bittered Cistanche puree prepared by the method not only reduces the bitterness, but also improves its sweetness and umami, making the overall taste richer. In addition, through enzymatic hydrolysis and Maillard reaction treatment, the content of echinacoside and verbascoside in the Cistanche puree is significantly increased, thereby improving its nutritional value.

[0010] Optionally, the composite protease is a protease with a pH value of 5.0 to 9.0.

[0011] Optionally, the composite protease is papain and flavor protease, and the ratio of the two is 1:3; the usage amount of the composite protease is 0.3-0.4g.

[0012] Optionally, in step (1), the solid-liquid ratio of the Cistanche deserticola to water is 1:20.

[0013] Optionally, in step (2), the enzymatic hydrolysis temperature is 45 to 55° C., and the enzymatic hydrolysis time is 1 to 2 h.

[0014] Optionally, in step (3), the pH value of the Cistanche deserticola hydrolysate is adjusted to 6-7.

[0015] Furthermore, the pH was adjusted to 6-7 with 2.0 mol / L NaOH and 1.0 mol / L HCl.

[0016] Optionally, in step (3), the temperature of the Maillard reaction is 110-130° C., the time is 15-30 min, and the stirring speed is 1000-2000 r / min.

[0017] On the other hand, the present invention provides a Cistanche deserticola puree based on the Maillard reaction obtained by the above preparation method, which reduces the bitterness of the Cistanche deserticola puree while enhancing its sweetness and umami taste, making the overall taste richer.

[0018] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The bitter peptide and bitter amino acid contents in the Cistanche deserticola puree according to the examples and comparative examples of the present invention;

[0020] Figure 2The contents of echinacoside and verbascoside in the Cistanche deserticola puree according to the examples and comparative examples of the present invention. DETAILED DESCRIPTION

[0021] The technical solution of the present invention is described below through specific examples. It should be understood that the one or more method steps mentioned in the present invention do not exclude the existence of other method steps before and after the combination step or the insertion of other method steps between these explicitly mentioned steps; it should also be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. Moreover, unless otherwise specified, the numbering of each method step is only a convenient tool for identifying each method step, and is not intended to limit the order of arrangement of each method step or to limit the scope of the present invention. The change or adjustment of the relative relationship thereof shall also be regarded as the scope of the present invention without substantially changing the technical content.

[0022] In order to better understand the above technical scheme, the exemplary embodiments of the present invention are described in more detail below. Although exemplary embodiments of the present invention are shown, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present invention and to enable the scope of the present invention to be fully communicated to those skilled in the art.

[0023] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0024] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are merely illustrative and do not limit the present invention in any way.

[0025] Example 1

[0026] A method for preparing desert cistanche puree based on Maillard reaction, comprising the following steps:

[0027] (1) Wash and cut fresh Cistanche deserticola into pieces; slurry Cistanche deserticola and water at a solid-liquid ratio of 1 g: 20 mL. Add 0.4 g of papain and flavor protease (at a ratio of 1:3, with papain and flavor protease activities of 10,000 U / g), adjust the pH of the enzymatic hydrolysis reaction to 7 using 2.0 mol / L NaOH and 1.0 mol / L HCl, place in a 45°C water bath, and perform enzymatic hydrolysis at 45°C for 2 h. Finally, inactivate the enzyme in 100°C boiling water for 10 min to obtain Cistanche deserticola enzymatic hydrolysate.

[0028] (2) Use 2.0 mol / L NaOH and 1.0 mol / L HCl to adjust the pH of the Cistanche deserticola hydrolyzate to 7; place it in a CNC super thermostatic bed, treat it at a constant temperature of 110°C, react for 20 minutes, and stir it at 1200 r / min. After the reaction is completed, cool it to room temperature to obtain the debittered Cistanche deserticola puree. Refrigerate it at 4°C for later use.

[0029] Example 2

[0030] A method for preparing desert cistanche puree based on Maillard reaction, comprising the following steps:

[0031] (1) Wash and cut fresh Cistanche deserticola into pieces; slurry Cistanche deserticola and water at a solid-liquid ratio of 1 g: 20 mL. Add 0.3 g of papain and flavor protease (at a ratio of 1:3, with papain and flavor protease activities of 10,000 U / g), adjust the pH of the enzymatic hydrolysis reaction to 6 using 2.0 mol / L NaOH and 1.0 mol / L HCl, place in a 50°C water bath, and perform enzymatic hydrolysis at 50°C for 2 h. Finally, inactivate the enzyme in 100°C boiling water for 15 min to obtain Cistanche deserticola enzymatic hydrolysate.

[0032] (2) Use 2.0 mol / L NaOH and 1.0 mol / L HCl to adjust the pH of the Cistanche deserticola hydrolysate to 6.5; place it in a CNC super thermostatic bed, treat it at a constant temperature of 110°C, react for 20 minutes, and stir it at 800 r / min. After the reaction is completed, cool it to room temperature to obtain the debittered Cistanche deserticola puree. Refrigerate it at 4°C for later use.

[0033] Example 3

[0034] A method for preparing desert cistanche puree based on Maillard reaction, comprising the following steps:

[0035] (1) Wash and cut fresh Cistanche deserticola into pieces; slurry Cistanche deserticola and water at a solid-liquid ratio of 1 g: 20 mL. Add 0.35 g of papain and flavor protease (at a ratio of 1:3, with papain and flavor protease activities of 10,000 U / g), adjust the pH of the enzymatic hydrolysis reaction to 7 using 2.0 mol / L NaOH and 1.0 mol / L HCl, place in a 45°C water bath, and perform enzymatic hydrolysis at 45°C for 2 h. Finally, inactivate the enzyme in 100°C boiling water for 10 min to obtain Cistanche deserticola enzymatic hydrolysate.

[0036] (2) Use 2.0 mol / L NaOH and 1.0 mol / L HCl to adjust the pH of the Cistanche deserticola hydrolyzate to 7; place it in a CNC super thermostatic bed, treat it at a constant temperature of 120°C, react for 25 minutes, and stir it at 1000 r / min. After the reaction is completed, cool it to room temperature to obtain the debittered Cistanche deserticola puree. Refrigerate it at 4°C for later use.

[0037] Comparative Example 1

[0038] Compared with Example 1, this comparative example provides a Cistanche deserticola puree that is not subjected to a composite protease enzymatic hydrolysis reaction to obtain an unenzymatically hydrolyzed Cistanche deserticola puree.

[0039] (1) Wash fresh Cistanche deserticola and cut it into pieces; pulp Cistanche deserticola and water at a solid-liquid ratio of 1 g: 20 mL, and adjust the pH of the unhydrolyzed Cistanche deserticola stock to 7 using 2.0 mol / L NaOH and 1.0 mol / L HCl. Place it in a CNC super thermostatic bed, treat it at a constant temperature of 110°C, react for 20 minutes, and stir at 1200 r / min. After the reaction is completed, cool it to room temperature to obtain the debittered Cistanche deserticola stock. Refrigerate it at 4°C for later use.

[0040] Comparative Example 2

[0041] Compared with Example 1, this comparative example only used papain to enzymolyze the obtained Cistanche deserticola puree.

[0042] (1) Wash fresh Cistanche deserticola and cut it into pieces; slurry Cistanche deserticola and water at a solid-liquid ratio of 1g:20mL. Add 0.4g of papain (papain activity is 10,000 U / g), adjust the pH of the enzymatic hydrolysis reaction to 7 with 2.0mol / L NaOH and 1.0mol / L HCl, put it in a 45℃ water bath, and enzymolyze it at 45℃ for 2h. Finally, inactivate the enzyme in 100℃ boiling water for 10min to obtain Cistanche deserticola enzymatic hydrolyzate.

[0043] (2) Use 2.0 mol / L NaOH and 1.0 mol / L HCl to adjust the pH of the Cistanche deserticola hydrolyzate to 7. Place it in a CNC super thermostatic bed and treat it at a constant temperature of 110°C for 20 minutes, stirring at 1200 r / min. After the reaction is completed, cool it to room temperature to obtain the debittered Cistanche deserticola puree. Refrigerate it at 4°C for later use.

[0044] Comparative Example 3

[0045] Compared with Example 1, in this comparative example, only flavor protease was used to enzymolyze the obtained Cistanche deserticola puree.

[0046] (1) Wash fresh Cistanche deserticola and cut it into pieces; slurry Cistanche deserticola and water at a solid-liquid ratio of 1g:20mL. Add 0.4g of flavor protease (flavor protease activity is 10,000 U / g), adjust the pH of the enzymatic hydrolysis reaction to 7 with 2.0mol / L NaOH and 1.0mol / L HCl, put it in a 45℃ water bath, and keep the enzymatic hydrolysis at 45℃ for 2h. Finally, inactivate the enzyme in 100℃ boiling water for 10min to obtain the Cistanche deserticola enzymatic hydrolysate.

[0047] (2) Use 2.0 mol / L NaOH and 1.0 mol / L HCl to adjust the pH of the Cistanche deserticola hydrolyzate to 7. Place it in a CNC super thermostatic bed and treat it at a constant temperature of 110°C for 20 minutes, stirring at 1200 r / min. After the reaction is completed, cool it to room temperature to obtain the debittered Cistanche deserticola puree. Refrigerate it at 4°C for later use.

[0048] Comparative Example 4

[0049] Compared with Example 1, the present comparative example did not carry out the Maillard reaction, and obtained the Cistanche deserticola puree enzymatic hydrolysate.

[0050] (1) Wash fresh Cistanche deserticola and cut it into pieces; beat Cistanche deserticola with water at a solid-liquid ratio of 1g:20mL. Add 0.4g of papain and flavor protease (at a ratio of 1:3, with papain and flavor protease activity of 10,000 U / g), adjust the pH of the enzymatic hydrolysis reaction to 7 with 2.0mol / L NaOH and 1.0mol / L HCl, place in a 45℃ water bath, and perform enzymatic hydrolysis at 45℃ for 2h. Finally, inactivate the enzyme in 100℃ boiling water for 10min to obtain Cistanche deserticola enzymatic hydrolysate.

[0051] Test example

[0052] Sensory evaluation team: 10 (5 males and 5 females) professionally trained and experienced R&D personnel were selected to form a sensory evaluation team, aged between 25 and 35 years old.

[0053] Application sample preparation: the debittered Cistanche puree prepared in Example 1, the non-enzymatically hydrolyzed Cistanche puree prepared in Comparative Example 1, the Cistanche puree hydrolyzed with a single enzyme prepared in Comparative Examples 2 and 3, and the Cistanche puree without Maillard reaction obtained in Comparative Example 4.

[0054] The samples to be evaluated were numbered and placed in a uniform container. The higher the total score of the sensory evaluation, the higher the acceptability. The final result was averaged. The evaluation team members were required not to drink alcohol or eat irritating food 12 hours before the evaluation. They were prohibited from discussing with each other during the evaluation process. After evaluating a sample, they rinsed their mouths with water and ate tasteless soda crackers to prevent the influence of samples. The evaluation time interval was 10 minutes. The results are shown in Table 1.

[0055] Table 1: Sensory Rating Table

[0056]

[0057] As can be seen from Table 1, the score of the Cistanche deserticola puree after composite enzymatic hydrolysis and Maillard reaction is significantly higher than that of the Cistanche deserticola puree of Comparative Examples 1-4, and the overall sensory score is 22. The Cistanche deserticola puree prepared in Example 1 has low bitterness, moderate sweetness, and a natural and rich taste.

[0058] The bitter peptides and bitter amino acids present in the Cistanche deserticola puree obtained in Example 1 and Comparative Examples 1-4 were measured.

[0059] (1) Pretreatment of the total amount of amino acids to be tested: 0.1 g of freeze-dried Cistanche deserticola puree sample was placed in a 15 mL hydrolysis tube, and then 8.0 mL of 6 mol / L HCl was added. After sealing under vacuum, the tube was placed in an oven at 110°C for hydrolysis for 24 h. After the hydrolysis was completed, the solution in the hydrolysis tube was fixed to 25 mL with ultrapure water, and then filtered with two layers of filter paper. 1.0 mL of the filtrate was placed in a 10 mL beaker, and vacuumed in a vacuum dryer for 30 min, and then dried overnight. The acid was evaporated, and the sample was taken out of the evaporated beaker, and 1.0 mL of 0.02 mol / L HCl was added. After standing for a while, the mixture was directly introduced into a 1.5 mL centrifuge tube, centrifuged at 10,000 r / min for 15 min, and stored at 4°C for testing.

[0060] Pretreatment of free amino acids to be tested: Take 1g of freeze-dried sample of Cistanche deserticola puree and dilute it to 50mL with trichloroacetic acid, let it stand for one hour, filter it with double-layer filter paper, take 1mL of the filtrate in a 1.5mL centrifuge tube, centrifuge it at 7000g / min for 10min, take 0.4mL of the supernatant, and store it at 4℃ for testing.

[0061] Agilent amino acid analysis column (column temperature 40 ° C; detection wavelength 338 nm; mobile phase flow rate 1 mL / min) was used, and the mobile phase composition included: Phase A, 8 g of crystalline sodium acetate was dissolved in 1 L of water, and then 225 μL of triethylamine was added, stirred and 5% acetic acid was added dropwise, the pH was adjusted to 7.20 ± 0.05, 5 mL of tetrahydrofuran was added, and the mixture was mixed for later use; Phase B: 12 ​​g of crystalline sodium acetate was weighed in an 800 mL beaker, 400 mL of water was added and stirred until all the crystalline water was dissolved, 5% acetic acid was added dropwise, the pH was adjusted to 7.20 ± 0.05, and the solution was added to 800 mL of acetonitrile and 800 mL of methanol, mixed and used for later use, and gradient elution was performed. The proportion of total bitter amino acids was analyzed.

[0062] (2) Using freeze-dried powder of Cistanche deserticola as the test object, a 20% (w / v) solution was prepared, and the same volume of isobutanol was added. The solution was stirred magnetically for 2 minutes, and then transferred to a 250 mL separatory funnel and allowed to stand. When the liquid was separated into two distinct layers, the aqueous phase liquid was discharged from the bottom of the funnel, and the aqueous phase peptide was obtained by concentration and freeze drying; the isobutanol residue was poured out from the funnel, and the alcohol phase peptide was obtained by concentration and freeze drying.

[0063] Analysis of bitter peptides in isobutanol extract by liquid chromatography-mass spectrometry Liquid chromatography setting conditions: Instrument Waters MALDI Synapt Q-TOF MS; BEH C18 column (1.7um, 2.1mm×150mm); Detection wavelength: 200-400nm. Injection volume: 5μL, flow rate: 0.3mL / min. Mobile phase: A: acetonitrile, B: formic acid / water (0.1%, V / V). Gradient elution conditions: 100% B (0-0.1min), 70% B (0.1-10min), 20% B (10-15min), 0% B (15-20min). Mass spectrometry setting conditions: ion mode: ESI+; capillary voltage: 3.5 kV; cone voltage: 30 V; mass range: 20-2000 m / z; ion source temperature: 100 °C; desolvation gas temperature: 400 °C; desolvation gas flow rate: 700 L / hr; cone backflush gas flow rate: 50 L / hr; collision energy: 6-20 eV; data were analyzed using MassLynx 4.1 software.

[0064] Specific results such as Figure 1 As shown, in this example, the content of bitter peptides and bitter amino acids in the Cistanche deserticola puree is reduced through composite enzymatic hydrolysis and Maillard reaction, thereby reducing the bitterness.

[0065] The contents of echinacoside and verbascoside in the Cistanche deserticola puree obtained in Example 1 and Comparative Examples 1-4 were measured.

[0066] Determination of echinacoside and verbascoside content:

[0067] Reference solution: Weigh 1 mg each of echinacoside reference and verbascoside reference in turn, place in a 5 ml volumetric flask, add 50% methanol to the mark, and prepare a 0.4 mg / mL reference solution.

[0068] Test solution: accurately weigh 1g of Cistanche freeze-dried powder and dried powder, place in a 50mL brown volumetric flask, accurately add 25mL of 50% methanol, seal, shake well, weigh, soak for 30min, treat with ultrasound (power 250W, frequency 40kHz) for 40min. Let cool to room temperature, weigh again, add 50% methanol to make up for the lost mass, and filter through a 0.22μm filter.

[0069] Chromatographic conditions: Octadecylsilane bonded silica gel was used as the filler column, mobile phase A was methanol, and mobile phase B was 0.1% formic acid solution.

[0070] Gradient conditions: 0-17 min: 26.5A: 73.5B; 17-20 min: 26.5→29.5A 73.5→70.5B; 20-47 min: 29.5A: 70.5B. The detection wavelength was 330 nm, the flow rate was 1.0 mL / min, the injection volume was 10 μL, and the column temperature was 30°C.

[0071] Take the mixed control sample and dilute it 2, 4, 10, 20, 40, and 80 times to obtain 6 different concentrations of reference solution, and measure it according to the above chromatographic conditions. Use the peak area as the ordinate and the concentration as the abscissa to perform linear regression and calculate the regression equation and linear range of each component.

[0072] Specific results such as Figure 2 As shown, in this example, the contents of echinacoside and verbascoside in Cistanche deserticola puree are significantly increased through composite enzymatic hydrolysis and Maillard reaction, thereby improving its nutritional value.

[0073] In summary, according to the embodiments of the present invention, the bitter peptides and bitter amino acids present in the Cistanche puree are reduced by composite enzymolysis and Maillard reaction, and the bitterness is reduced. Therefore, compared with the traditional Cistanche puree, the de-bittered Cistanche puree prepared by the present method not only reduces the bitterness, but also improves its sweetness and umami, making the overall taste richer. In addition, through enzymolysis and Maillard reaction treatment, the content of echinacoside and verbascoside in the Cistanche puree is significantly increased, thereby improving its nutritional value.

[0074] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms should not be understood as necessarily being directed to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine different embodiments or examples described in this specification.

[0075] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for preparing Cistanche deserticola puree based on Maillard reaction, characterized in that: The following steps are involved: (1) adding water to the Cistanche deserticola to make a pulp, thereby obtaining a Cistanche deserticola pulp that has not been debittered; (2) adding a composite protease to the undebittered Cistanche deserticola puree for enzymolysis to obtain a Cistanche deserticola enzymolysis solution; (3) adjusting the pH value of the Cistanche deserticola enzymatic hydrolyzate, heating and stirring to perform Maillard reaction, and cooling to obtain debittered Cistanche deserticola puree.

2. The preparation method according to claim 1, characterized in that The composite protease is a protease with a pH value of 5.0 to 9.

0.

3. The preparation method according to claim 1 or 2, characterized in that: The composite protease is papain and flavor protease, and the ratio of the two is 1:3; the usage amount of the composite protease is 0.3-0.4g.

4. The preparation method according to claim 1, characterized in that: In step (1), the solid-liquid ratio of the Cistanche deserticola to water is 1:

20.

5. The preparation method according to claim 1, characterized in that: In step (2), the enzymatic hydrolysis temperature is 45 to 55° C. and the enzymatic hydrolysis time is 1 to 2 h.

6. The preparation method according to claim 1, characterized in that: In step (3), the pH value of the Cistanche deserticola hydrolyzate is adjusted to 6-7.

7. The preparation method according to claim 6, characterized in that: The pH was adjusted to 6-7 by 2.0 mol / L NaOH and 1.0 mol / L HCl.

8. The preparation method according to claim 1, characterized in that: In step (3), the temperature of the Maillard reaction is 110-130° C., the time is 15-30 min, and the stirring speed is 1000-2000 r / min.

9. A Cistanche deserticola puree based on the Maillard reaction, characterized in that: The method is prepared according to any one of claims 1 to 8.

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