Preparation method of orange dietary fiber as well as product and application of orange dietary fiber
By adopting enzymatic reaction and ethanol precipitation during the extraction process of orange dietary fiber, the problems of high production cost and poor stability of dietary fiber in the prior art are solved, and orange dietary fiber with excellent adsorption and inhibitory activity are prepared, which is suitable for a variety of application fields.
Patent Information
- Application Number
- CN202510195227.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-21
- Publication Date
- 2025-06-20
AI Technical Summary
In the prior art, the production cost of dietary fiber is high, the product stability is poor, and the active ingredients of orange dietary fiber are less studied, and there is a lack of low-cost, simple process and stable extraction methods.
Orange dietary fiber is prepared by homogenizing the orange pulp and adding water to the enzyme reaction, and the complex enzyme extraction method of neutral protease and cellulase is performed to perform enzymatic decomposition, precipitation, redissolution, dialysis and drying.
The prepared orange pulp soluble fiber extract has excellent adsorption nitrite activity and inhibits cholesterol esterase activity, and has good glucose adsorption ability and excellent water retention. It is suitable for food industry and dietary fiber supplements.
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Figure CN120167635A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of nutritional products, and particularly relates to a preparation method, product and application of orange dietary fiber. Background Art
[0002] Dietary fiber is an important class of nutrients. Existing studies have shown that dietary fiber not only helps to promote intestinal health, but also can improve blood glucose control and reduce cholesterol levels, playing an important role in preventing various chronic diseases.
[0003] Dietary fiber includes soluble dietary fiber (SDF) and insoluble dietary fiber (IDF). Dietary fiber has broad application prospects in the food industry. It can be used as a natural food additive to enhance the taste and texture of food; it can also be added to products such as beverages and yogurts to increase the fiber content and nutritional value of the products. In addition, dietary fiber can be used to make low-calorie, high-fiber foods to meet consumers' demand for healthy foods.
[0004] As a dietary fiber supplement, dietary fiber also has broad market prospects in the health product industry. It can help people increase their intake of dietary fiber and promote intestinal health; it can also be used as an auxiliary ingredient in weight loss products to help consumers control their weight.
[0005] However, during the extraction process of dietary fiber, the active components of dietary fiber may be damaged, which reduces its nutritional value and health functions. In the prior art, the production cost of dietary fiber is relatively high and the product stability is poor, which limits its wide application.
[0006] Oranges are rich in dietary fiber, but there is little research on the dietary fiber in orange pulp, and the activity of orange dietary fiber needs to be further studied. Developing a method for extracting orange dietary fiber with low cost, simple process and better stability and studying its functional activity are technical problems to be solved. Summary of the Invention
[0007] The purpose of this part is to outline some aspects of the embodiments of the present invention and briefly introduce some preferred embodiments.
[0008] As one aspect of the present invention, the present invention provides a preparation method of orange dietary fiber, which includes homogenizing orange pulp, adding water, sequentially adding neutral protease and cellulase for enzymatic hydrolysis reaction, centrifuging, taking the supernatant, adding an ethanol solution with a volume concentration of 70-75% for precipitation, centrifuging to collect the precipitate, redissolving with water, dialyzing, and drying to obtain orange dietary fiber.
[0009] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: adding water after homogenizing the orange pulp means adding 12 - 15 times the volume of water after homogenizing the orange pulp.
[0010] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: adding neutral protease and cellulase in sequence for enzymatic hydrolysis reaction means first adjusting the pH to 6.5 - 6.8, adding neutral protease, hydrolyzing at a temperature of 40 - 45°C for 0.5 - 2 h, then adjusting the pH to 4.5 - 5, adding cellulase, and hydrolyzing at a temperature of 50 - 55°C for 2 - 3 h.
[0011] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: the activity concentration of the neutral protease is 1 - 2.4 U / mL; the activity concentration of the cellulase is 2 - 20 U / mL.
[0012] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: the volume concentration of the ethanol solution is 71 - 74%.
[0013] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: adding an ethanol solution with a volume concentration of 70 - 75% for precipitation means precipitating at a temperature of 2 - 6°C for 12 - 24 h.
[0014] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: adding water for re - dissolution means adding 10 - 15 times the mass of the precipitate of water for re - dissolution; the dialysis is carried out with a 20 kDa dialysis bag for 48 - 72 h.
[0015] As a preferred embodiment of the preparation method of the orange dietary fiber described in the present invention: the drying is carried out by vacuum freeze - drying at - 40°C for 24 - 48 h.
[0016] The present invention also provides the orange dietary fiber prepared by the preparation method of the orange dietary fiber described above.
[0017] Advantages of the present invention: The soluble fiber extract of orange pulp prepared by the present invention has excellent activities of adsorbing nitrite and inhibiting cholesterol esterase. The composite enzyme extraction method using protease and cellulase has better adsorption activity compared with the products extracted by alkali extraction or acid extraction, and at the same time has higher activity of inhibiting cholesterol esterase. Among them, the final concentration of ethanol affects the structure and activity of the extracted fiber. In addition, the orange fiber extract of the present invention also has good glucose adsorption ability and excellent water - holding capacity. The orange fiber extract of the present invention can be used in the food industry, such as beverages, baked foods, dairy products, etc., as well as in the fields of dietary fiber supplements, pharmaceutical excipients, etc., and can assist in reducing blood lipids and blood sugar, adsorbing harmful substances, and being beneficial to intestinal health. The preparation method of the present invention is simple and has a wide application prospect. Description of the Drawings
[0018] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for the description of the embodiments will be briefly introduced below, where:
[0019] Figure 1 Test results of the nitrite adsorption rate of the orange fiber extract prepared for each embodiment.
[0020] Figure 2 Test results of the inhibition rate of the orange fiber extract prepared for each embodiment on cholesterol esterase activity.
[0021] Figure 3 Test results of the water retention of the orange fiber extract prepared for each embodiment. Specific embodiments
[0022] To make the above objects, features, and advantages of the present invention more obvious and understandable, the specific embodiments of the present invention will be described in detail below in conjunction with specific embodiments.
[0023] Example 1:
[0024] Peel the oranges and discard the peels. Homogenize the orange pulp (3000 r / min, 2 min), add deionized water with a volume 15 times that of the pulp slurry and mix evenly. Adjust the pH to 6.8, add neutral protease (CAS No.: 9068-59-1) at 2.4 U / mL, and enzymatically hydrolyze at 45 °C for 1 h. Adjust the pH to 4.5, add cellulase (CAS No.: 9012-54-8) at 10 U / mL, and enzymatically hydrolyze at 50 °C for 2 h. Centrifuge at 200 r / min for 10 min, collect the supernatant, add 95% ethanol with a volume 3 times that of the supernatant, precipitate at 4 °C for 12 h, centrifuge at 800 r / min for 10 min to collect the precipitate, dissolve the precipitate in deionized water with a mass 10 times that of the precipitate, dialyze with a 20 kDa dialysis bag for 48 h, and then vacuum freeze-dry at -40 °C for 24 h to obtain the orange fiber extract.
[0025] Prepare a 150 mg / L sodium nitrite aqueous solution, and dilute it to 0, 5 mg / L, 10 mg / L, 20 mg / L, 30 mg / L, and 40 mg / L respectively. Take 1 mL of each diluted solution, add 2.5 mL of 60 wt% acetic acid, then add 2.5 mL of 10 g / L sulfanilic acid and 2.5 mL of 1 g / L N-1-naphthylethylenediamine. Add deionized water to make the volume up to 25 mL. After standing in the dark for 25 min, measure the absorbance at 550 nm and draw the nitrite standard curve. Take 0.15 g of orange fiber extract sample and add it to 25 mL of 20 mg / L sodium nitrite aqueous solution. Adjust the pH to 2, place it in a constant temperature shaker at 37 °C. Take out the sample at 30 min and 90 min of adsorption respectively, centrifuge at 4000 r / min for 20 min. Take 1 mL of the supernatant, add 2.5 mL of 60 wt% acetic acid, then add 2.5 mL of 10 g / L sulfanilic acid and 2.5 mL of 1 g / L N-naphthylethylenediamine. Add deionized water to make the volume up to 25 mL. After standing in the dark for 25 min, measure the nitrite adsorption rate at 30 min and 90 min of adsorption respectively at 550 nm.
[0026] Prepare a 0.1 mol / L phosphate buffer solution with pH = 7.0 (containing 0.1 mol / L sodium chloride and 5.16 mmol / L sodium taurocholate), add 0.2 mmol / L cholesteryl palmitate and 1 mg / mL orange fiber extract sample, mix at 37 °C for 10 min, add 1000 U / L cholesterol esterase, react at 37 °C for 10 min, measure the absorbance at 405 nm, and calculate the inhibition rate of the sample on cholesterol esterase activity. The blank control is without adding the orange fiber extract sample.
[0027] Water retention test: Accurately weigh 10 g of orange fiber extract, place it in an environment with 90% air humidity for 24 h, take it out and weigh it. Then place it in a dry environment with air humidity below 20%, and weigh it again after 48 h. Calculate the weight loss rate. The weight loss rate = ((the weight of the second weighing - the weight of the first weighing) / the weight of the first weighing) × 100%. The measured weight loss rate is 3.15%.
[0028] Research Example 1:
[0029] Peel the oranges and discard the peels. Homogenize the orange pulp, add deionized water with a volume 15 times that of the pulp slurry, mix evenly, adjust the pH to 6.8, add neutral protease (CAS No.: 9068-59-1) at 2.4 U / mL, enzymatically hydrolyze at 45 °C for 1 h, adjust the pH to 4.5, add cellulase (CAS No.: 9012-54-8) at 10 U / mL, enzymatically hydrolyze at 50 °C for 2 h, centrifuge at 200 r / min for 10 min, collect the supernatant, add 95% ethanol with a volume 5 times that of the supernatant, precipitate at 4 °C for 12 h, centrifuge at 800 r / min for 10 min to collect the precipitate, redissolve with deionized water with a mass 10 times that of the precipitate, dialyze with a 20 kDa dialysis bag for 48 h, and then vacuum freeze-dry at -40 °C for 24 h to obtain the orange fiber extract.
[0030] Research Example 2:
[0031] Peel the oranges and discard the peels. Homogenize the orange pulp, add deionized water with a volume 15 times that of the pulp slurry, mix evenly, adjust the pH to 6.8, add neutral protease (CAS No.: 9068-59-1) at 2.4 U / mL, enzymatically hydrolyze at 45 °C for 1 h, adjust the pH to 4.5, add cellulase (CAS No.: 9012-54-8) at 10 U / mL, enzymatically hydrolyze at 50 °C for 2 h, centrifuge at 200 r / min for 10 min, collect the supernatant, add 95% ethanol with a volume 1.5 times that of the supernatant, precipitate at 4 °C for 12 h, centrifuge at 800 r / min for 10 min to collect the precipitate, redissolve with deionized water with a mass 10 times that of the precipitate, dialyze with a 20 kDa dialysis bag for 48 h, and then vacuum freeze-dry at -40 °C for 24 h to obtain the orange fiber extract.
[0032] Research Example 3:
[0033] Peel the oranges and discard the peels. Homogenize the orange pulp, add a hydrochloric acid aqueous solution with a pH of 2 and a volume 15 times that of the pulp slurry, magnetically stir at 40 °C for 2 h, centrifuge at 200 r / min for 10 min, collect the supernatant, adjust to pH = 7 with sodium hydroxide, add 95% ethanol with a volume 3 times that of the supernatant, precipitate at 4 °C for 12 h, centrifuge at 800 r / min for 10 min to collect the precipitate, redissolve with deionized water with a mass 10 times that of the precipitate, dialyze with a 20 kDa dialysis bag for 48 h, and then vacuum freeze-dry at -40 °C for 24 h to obtain the orange fiber extract.
[0034] Research Example 4:
[0035] Peel the oranges and discard the peels. Homogenize the orange pulp, add an aqueous solution of Na2CO3 with a mass concentration of 5% at 15 times the volume of the pulp slurry, stir magnetically at 40 °C for 2 h, centrifuge at 200 r / min for 10 min, collect the supernatant, adjust to pH = 7 with hydrochloric acid, add 95% ethanol at 3 times the volume of the supernatant, precipitate at 4 °C for 12 h, centrifuge at 800 r / min for 10 min to collect the precipitate, dissolve the precipitate in deionized water at 10 times the mass, dialyze with a 20 kDa dialysis bag for 48 h, and then vacuum freeze-dry at -40 °C for 24 h to obtain the orange fiber extract.
[0036] The test results of the nitrite adsorption rate of the orange fiber extracts prepared in each example are shown in Table 1 and Figure 1 .
[0037] Table 1
[0038] Adsorption rate at 30 min (%) Adsorption rate at 90 min (%) Example 1 45.4％ 95.1％ Research Example 1 36.2％ 81.4％ Research Example 2 31.7％ 77.3％ Research Example 3 25.8％ 61.2％ Research Example 4 26.1％ 68.6％
[0039] The test results of the inhibition rate of the cholesterol esterase activity of the orange fiber extracts prepared in each example are shown in Table 2 and Figure 2 .
[0040] Table 2
[0041]
[0042] The test results of the water retention of the orange fiber extracts prepared in each example are shown in Table 3 and Figure 3 .
[0043] Table 3
[0044] Weight loss rate (%) Example 1 3.15％ Research Example 1 3.42％ Research Example 2 3.79％ Research Example 3 6.71％ Research Example 4 4.36％
[0045] Comparative Example 1:
[0046] Take the orange peels, dry and crush them, pass through a 60-mesh sieve, add deionized water at 15 times the volume and mix evenly, adjust the pH = 6.8, add neutral protease (CAS No.: 9068-59-1) at 2.4 U / mL, enzymatically hydrolyze at 45 °C for 1 h, adjust the pH = 4.5, add cellulase (CAS No.: 9012-54-8) at 10 U / mL, enzymatically hydrolyze at 50 °C for 2 h, centrifuge at 200 r / min for 10 min, collect the supernatant, add 95% ethanol at 3 times the volume of the supernatant, precipitate at 4 °C for 12 h, centrifuge at 800 r / min for 10 min to collect the precipitate, dissolve the precipitate in deionized water at 10 times the mass, dialyze with a 20 kDa dialysis bag for 48 h, and then vacuum freeze-dry at -40 °C for 24 h to obtain the orange fiber extract.
[0047] Experimental results: The nitrite adsorption rates at 30 min and 90 min were tested according to the method of Example 1 and were 23.6% and 57.0% respectively.
[0048] The inhibition rate of cholesterol esterase activity was 58%.
[0049] The weight loss rate of the water retention test sample was 5.47%.
[0050] In summary, the orange pulp soluble fiber extract prepared by the present invention has excellent nitrite adsorption activity and cholesterol esterase inhibition activity. The composite enzyme extraction method using protease and cellulase has better adsorption activity than the products extracted by alkali or acid, and at the same time has higher cholesterol esterase inhibition activity. Among them, the final ethanol concentration affects the structure and activity of the extracted fiber. In addition, the orange fiber extract of the present invention also has good glucose adsorption ability and excellent water retention. The orange fiber extract of the present invention can be used in the food industry, such as beverages, baked foods, dairy products, etc., as well as in the fields of dietary fiber supplements, pharmaceutical excipients, etc., and can assist in reducing blood lipids and blood sugar, adsorbing harmful substances, and being beneficial to intestinal health. The preparation method of the present invention is simple and has a wide application prospect.
[0051] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention, and they should all be covered within the scope of the claims of the present invention.
Claims
1. A method for preparing orange dietary fiber, characterized in that: The method comprises the steps of homogenizing orange pulp, adding water, sequentially adding neutral protease and cellulase for enzymolysis reaction, centrifuging, taking supernatant, adding ethanol solution with a volume concentration of 70-75% for precipitation, centrifuging and collecting precipitation, adding water for redissolution, dialyzing, and drying to obtain orange dietary fiber.
2. The method for preparing orange dietary fiber according to claim 1, characterized in that: The step of adding water after homogenizing the orange pulp comprises adding 12 to 15 times the volume of water after homogenizing the orange pulp.
3. The method for preparing orange dietary fiber according to claim 1 or 2, characterized in that: The neutral protease and cellulase are added sequentially for enzymatic hydrolysis reaction by first adjusting the pH to 6.5-6.8, adding the neutral protease, and performing enzymatic hydrolysis at 40-45°C for 0.5-2h, then adjusting the pH to 4.5-5, adding the cellulase, and performing enzymatic hydrolysis at 50-55°C for 2-3h.
4. The method for preparing orange dietary fiber according to claim 3, characterized in that: The activity concentration of the neutral protease is 1-2.4 U / mL; the activity concentration of the cellulase is 2-20 U / mL.
5. The method for preparing orange dietary fiber according to claim 1 or 2, characterized in that: The ethanol solution (volume concentration is 71-74%).
6. The method for preparing orange dietary fiber according to claim 1 or 2, characterized in that: The precipitation of adding ethanol solution with a volume concentration of 70-75% is carried out at a temperature of 2-6° C. for 12-24 hours.
7. The method for preparing orange dietary fiber according to claim 1 or 2, characterized in that: The water-adding re-dissolution is to add water 10 to 15 times the mass of the precipitate for re-dissolution; and the dialysis is to use a 20 kDa dialysis bag for 48 to 72 hours.
8. The method for preparing orange dietary fiber according to claim 1 or 2, characterized in that: The drying is performed by vacuum freeze drying at -40°C for 24 to 48 hours.
9. Use of the orange dietary fiber prepared by the preparation method of orange dietary fiber according to claim 1 in food and / or pharmaceutical additives.
10. Use of the orange dietary fiber prepared by the preparation method of orange dietary fiber according to claim 1 in preparing food and / or pharmaceutical additives having the functions of absorbing nitrite, inhibiting cholesterol esterase activity and retaining water.