Kale composite fruit and vegetable enzyme and preparation method thereof

Abstract

The invention belongs to the field of preparation of fruit and vegetable enzymes, and particularly relates to a kale composite fruit and vegetable enzyme and a preparation method thereof. According to the present invention, the crushed mixed raw material and the mold agent are subjected to mixed fermentation, the fruit and vegetable enzyme liquid obtained through filtration is easily absorbed by the body, the viscous compound formed through cross-linking completely adsorbs beta-ionone and vanillin acetate, and the obtained collard compound enzyme liquid prepared from the prepared flavor adjusting nano-particle mixed fruit and vegetable enzyme liquid has characteristics of fine taste and good taste; hydrogen bonds are formed between the propylene glycol alginate and the sorbitan tristearate, the formed compound solution wraps the nutritional slurry prepared after grinding, and the obtained nutritional supplement composition is a stable suspension; the activity maintenance agent can effectively maintain the activity of the extracted fruit and vegetable enzyme, and when the activity maintenance agent is mixed with the kale compound enzyme liquid and the nutritional supplement composition, the prepared kale compound fruit and vegetable enzyme is stable in effect, not prone to losing activity in the storage process, rich in nutritional ingredient and excellent in flavor and taste.

Description

Technical Field

[0001] The invention belongs to the field of fruit and vegetable enzyme preparation, and specifically relates to a kale composite fruit and vegetable enzyme and a preparation method thereof. Background Art

[0002] As a "superfood", kale is rich in vitamins, minerals, antioxidants and dietary fiber, and has extremely high nutritional value. However, some nutrients in kale are difficult to be directly absorbed by the human body, which limits its effectiveness. Enzymatic hydrolysis technology can effectively decompose plant cell walls, release more active substances, and improve the bioavailability of nutrients.

[0003] Kale fruit and vegetable enzyme is a functional food made from kale as the main raw material, combined with other fruit and vegetable ingredients, through enzymatic hydrolysis and other processes. During the preparation process, the kale is usually washed, crushed, and made into a slurry, and then blanched to inactivate the chlorophyllase to maintain its green color. Subsequently, pectinase and protease are added for enzymatic hydrolysis to make the nutrients easier for the human body to absorb. This composite fruit and vegetable enzyme not only retains the nutritional components of kale, but also integrates the flavors of other fruits and vegetables. The final product can be used in beverages, health care products and other fields to meet consumers' demand for healthy food.

[0004] At present, the existing fruit and vegetable enzyme preparation technology has the following problems: first, the existing fruit and vegetable enzymes are easily lost in activity during processing and storage, affecting their efficacy; second, the nutrients in fruits and vegetables are destroyed during processing, and the nutrient content of the prepared fruit and vegetable enzymes is low; third, the existing fruit and vegetable enzyme preparation technology produces bitter or unpleasant flavor substances during the enzymatic hydrolysis process, affecting the taste of the fruit and vegetable enzymes. Summary of the invention

[0005] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides a kale composite fruit and vegetable enzyme and a preparation method thereof. In order to solve the problems that the existing fruit and vegetable enzymes are easy to lose activity, the efficacy decreases, the nutrient content is low, and the flavor is poor, the present invention uses a kale composite enzyme liquid, an activity maintaining agent, and a nutritional supplement composition to mix. The prepared kale composite fruit and vegetable enzyme has stable efficacy, rich nutrients, and good flavor.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is as follows: the present invention proposes a kale composite fruit and vegetable enzyme, and the raw materials for preparing the kale composite fruit and vegetable enzyme specifically include the following components in parts by weight: 40-60 parts of kale complex enzyme liquid, 15-19 parts of nutritional supplement composition, 0.8-1.2 parts of magnesium sulfate, 1.1-1.3 parts of calcium chloride, 0.6-0.9 parts of zinc sulfate, and 0.8-1.2 parts of L-cysteine.

[0007] Preferably, the raw materials for preparing the kale complex enzyme solution include the following components in parts by weight: 23-29 parts of kale, 16-18 parts of grapefruit, 19-22 parts of prunes, 7-11 parts of spinach, 6-10 parts of arugula, 11-13 parts of Parmesan cheese, 12-14 parts of quinoa, 1.4-1.7 parts of fungal agent, 14-17 parts of pullulan, 15-21 parts of polyglycerol ricinoleate, 0.8-1.4 parts of β-ionone, and 0.9-1.3 parts of vanillin acetate.

[0008] Preferably, the raw materials for preparing the nutritional supplement composition include the following components in parts by weight: 8-10 parts of chickpeas, 12-15 parts of pecan kernels, 6-10 parts of red snapper, 11-13 parts of propylene glycol alginate, 10-12 parts of sorbitan tristearate, and 0.7-1.1 parts of sodium hydroxide.

[0009] Preferably, the method for preparing the kale complex enzyme solution specifically comprises the following steps: S1. Put kale, grapefruit, prune, spinach, arugula, Parmesan cheese and quinoa into a grinder with a power of 2.2-2.6 kW, grind at a temperature of 20-26° C., for 18-26 min, and at a grinding speed of 3000-4000 r / min to obtain a mixed raw material; S2, placing the mold inoculant and 5% sterile glucose solution in a constant temperature shaking incubator with a power of 2.2-2.4 kW, the constant temperature shaking incubation temperature is 28°C, the constant temperature shaking incubation time is 24-30 h, the constant temperature shaking incubation speed is 220-240r / min, and the constant temperature shaking incubation is performed to obtain a mold solution; S3, placing the mixed raw material prepared by S1 and the mold liquid prepared by S2 into a fermentation tank with a power of 2.2-2.6 kW, fermenting at a temperature of 28-33° C., a fermentation speed of 90-110 r / min, and a fermentation time of 35-42 d, and filtering with a sterile filter membrane with a pore size of 0.22 mm after fermentation to obtain a fruit and vegetable enzyme liquid; S4, placing pullulan, polyglycerol ricinoleate, ultrapure water and rapeseed oil in a thermostatic agitator with a power of 1.6-1.9 kW, with a constant temperature of 38-55°C, a constant speed of 300-500 r / min, a constant temperature of 12-16 h, and reacting under constant temperature stirring to obtain a viscous compound; S5, placing β-ionone, vanillin acetate, and anhydrous ethanol in a stirrer with a power of 1.5-1.8 kW, stirring at a temperature of 25-33°C, a stirring speed of 300-320 r / min, and a stirring time of 10-15 min, adding the viscous compound prepared by S4 after dissolution, stirring at a temperature of 25-33°C, a stirring speed of 300-320 r / min, and a stirring time of 18-22 min, fully adsorbing, placing in a vacuum freeze nano spray dryer with a power of 2.3-2.8 kW, a vacuum degree of -0.05 MPa, a freeze nano spray drying temperature of -40°C, a freeze nano spray drying particle size of 100 nm, and a freeze nano spray drying time of 2.5-3 h, and freeze spray drying to obtain flavor-adjusting nanoparticles; S6. Put the flavor regulating nanoparticles prepared in S5 and the fruit and vegetable enzyme liquid prepared in S3 into a stirrer with a power of 1.5-1.8 kW, stir at a temperature of 20-26°C, a stirring speed of 300-400 r / min, and a stirring time of 20-30 min, mix them evenly to obtain a kale complex enzyme liquid.

[0010] Further, in S2, the mold agent is composed of Streptomyces herbicola, Mucor circinelloides, and Penicillium oxalicum, with a weight ratio of 1.1:0.9:1. The Streptomyces herbicola was purchased from the China General Microbiological Culture Collection Center with a preservation number of CGMCC4.6834, the Mucor circinelloides was purchased from the China General Microbiological Culture Collection Center with a preservation number of CGMCC3.7107, and the Penicillium oxalicum was purchased from the China General Microbiological Culture Collection Center with a preservation number of CGMCC3.15651. The solid-liquid ratio of the mold agent to 5% sterile glucose solution is 1:100 g / mL.

[0011] Furthermore, in S4, the mass fraction of the pullulan in ultrapure water is 8-12%, and the mass fraction of polyglycerol ricinoleate in rapeseed oil is 6-9%.

[0012] Furthermore, in S5, the mass fraction of the β-ionone in anhydrous ethanol is 1.7-2.6%.

[0013] Preferably, the method for preparing the nutritional supplement composition comprises the following steps: L1. Put chickpeas, pecan kernels and red snapper into a pulping machine with a power of 1.8-2.4 kW, the pulping temperature is 85-90°C, the pulping time is 35-40 min, the pulping speed is 18000-20000 r / min, and the pulping is performed to obtain a nutrient slurry; L2. Place propylene glycol alginate, sorbitan tristearate, sodium hydroxide, ultrapure water, and olive oil in a constant temperature stirrer with a power of 1.6-1.9 kW, with a constant temperature stirring temperature of 60-70°C, a constant temperature stirring speed of 300-500 r / min, and a constant temperature stirring time of 6-8 h to react and form a complex solution; L3. Place the nutrient slurry prepared by L1 and the complex solution prepared by L2 into a high-pressure homogenizer with a power of 1.8 kW, a pressure of 20 MPa, a high-pressure homogenization temperature of 60°C, a high-pressure homogenization time of 12-15 min, and a high-pressure homogenization speed of 1000-2000 r / min to obtain a nutritional supplement composition.

[0014] The present invention also proposes a method for preparing a kale composite fruit and vegetable enzyme, which specifically comprises the following steps: Step 1, put magnesium sulfate, calcium chloride, zinc sulfate, L-cysteine, and ultrapure water into a stirrer with a power of 1.5-1.8 kW, stir at a temperature of 28-33° C., stir at a speed of 400-600 r / min, stir for 12-18 min, and stir to obtain an active maintenance agent; Step 2: Add the kale complex enzyme solution to the activity maintaining agent prepared in step 1, stir at a temperature of 28-33° C., a stirring speed of 400-600 r / min, and a stirring time of 15-20 min, and mix well to obtain the kale enzyme solution; Step 3: Add the nutritional supplement composition to the kale enzyme solution prepared in step 2, use an ultrasonic agitator with a power of 1.8 kW, an ultrasonic frequency of 15 kHz, a stirring speed of 20,000 r / min, a stirring temperature of 25°C, and a stirring time of 10 min, and obtain the kale composite fruit and vegetable enzyme after uniform dispersion.

[0015] Preferably, in step 1, the mass fraction of magnesium sulfate in ultrapure water is 1.4%.

[0016] The beneficial effects achieved by the present invention are as follows: The invention adopts kale, grapefruit, prune, spinach, rocket, Parmesan cheese and quinoa to be crushed to obtain a mixed raw material rich in vitamins, minerals, dietary fiber, antioxidants and protein, and the mixed raw material is fermented with a mold agent containing herbicidal streptomyces, circinelloides and penicillium oxalicum, so that the anti-nutritional factors in the plant materials are decomposed, and the fruit and vegetable enzyme liquid obtained after filtration contains small molecular substances that are more easily absorbed by the body, and the obtained fruit and vegetable enzyme liquid has high bioavailability; pullulan and polyglycerol ricinoleate are cross-linked through hydrogen bonds to form a network structure, and the obtained viscous compound fully absorbs β-ionone and vanillin acetate with aromatic and sweet odors; after freeze spray drying, the obtained flavor regulating nanoparticles can be uniformly dispersed in the fruit and vegetable enzyme liquid to form a stable suspension system, thereby improving the smoothness and softness of the fruit and vegetable enzyme liquid, making it have better viscosity and fluidity, and the kale composite enzyme liquid obtained after mixing has a delicate taste; chickpeas, pecan kernels and red snapper are mixed and slurried to obtain a fat-containing A nutrient slurry containing propylene glycol alginate and sorbitan tristearate is prepared, and then stirred. The carboxyl group of propylene glycol alginate and the sorbitan group of sorbitan tristearate can form hydrogen bonds. The formed complex solution can form a stable interface layer on the surface of the nutrient slurry to prevent it from directly contacting the external environment. This encapsulation effect can prevent the nutrients from being degraded or precipitated prematurely during the digestion process, thereby improving their solubility and dispersibility in the digestive tract. The obtained nutritional supplement composition is a stable suspension. Magnesium sulfate, calcium chloride, zinc sulfate, and L-cysteine ​​are dissolved in ultrapure water. The obtained activity maintaining agent can effectively maintain the activity of the extracted fruit and vegetable enzymes by providing metal ions, maintaining the structural stability of the fruit and vegetable enzymes, and providing a reducing environment. The activity maintaining agent is mixed with the kale composite enzyme liquid and the nutritional supplement composition. The prepared kale composite fruit and vegetable enzyme has stable efficacy, is not easy to lose activity during storage, is rich in nutrients, and has excellent flavor and taste. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the present invention or the technical solutions in the prior art, the following will be described in a clear and understandable manner in conjunction with the accompanying drawings. Obviously, the drawings described below are only for the present invention, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0018] Figure 1 This is a scanning electron microscope image of the internal structure of the viscous compound of the present invention; Figure 2 This is a graph showing the first black discharge time of each group of mice in Experimental Example 1 of the present invention; Figure 3 This is a result graph showing the total amount of black feces discharged by each group of mice within 5 hours as described in Experimental Example 1 of the present invention; Figure 4 This is a graph showing the intestinal propulsion rate of mice described in Experimental Example 1 of the present invention; Figure 5 This is a picture of the finished product of the kale composite fruit and vegetable enzyme of the present invention. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments; based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0020] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only and are not intended to limit the content of this application.

[0021] The experimental methods in the following examples are conventional methods unless otherwise specified; the experimental materials and test strains used in the following examples are purchased from commercial channels unless otherwise specified.

[0022] Example 1: This example provides a kale composite fruit and vegetable enzyme and a preparation method thereof. The kale composite fruit and vegetable enzyme comprises the following components in parts by weight: 40 parts of kale complex enzyme liquid, 15 parts of nutritional supplement composition, 0.8 parts of magnesium sulfate, 1.1 parts of calcium chloride, 0.6 parts of zinc sulfate, and 0.8 parts of L-cysteine.

[0023] The raw materials for preparing the kale complex enzyme solution include the following components in parts by weight: 23 parts of kale, 16 parts of grapefruit, 19 parts of prunes, 7 parts of spinach, 6 parts of rocket, 11 parts of Parmesan cheese, 12 parts of quinoa, 1.4 parts of mold agent, 14 parts of pullulan, 15 parts of polyglycerol ricinoleate, 0.8 parts of β-ionone, and 0.9 parts of vanillin acetate.

[0024] The raw materials for preparing the nutritional supplement composition include the following components in parts by weight: 8 parts of chickpeas, 12 parts of pecan kernels, 6 parts of red snapper, 11 parts of propylene glycol alginate, 10 parts of sorbitan tristearate, and 0.7 parts of sodium hydroxide.

[0025] The preparation method of the kale complex enzyme solution specifically comprises the following steps: S1. Put kale, grapefruit, prune, spinach, arugula, Parmesan cheese, and quinoa into a 2.2 kW grinder at a grinding temperature of 20° C., a grinding time of 18 min, and a grinding speed of 3000 r / min to obtain a mixed raw material; S2, placing the mold inoculant and 5% sterile glucose solution in a constant temperature shaking incubator with a power of 2.2 kW, the constant temperature shaking incubation temperature is 28°C, the constant temperature shaking incubation time is 24 h, the constant temperature shaking incubation speed is 220 r / min, and the constant temperature shaking incubation is performed to obtain a mold solution; S3, putting the mixed raw material prepared by S1 and the mold liquid prepared by S2 into a fermentation tank with a power of 2.2 kW, fermenting at a temperature of 28°C, a fermentation speed of 90 r / min, and a fermentation time of 35 days, and filtering with a sterile filter membrane filter with a pore size of 0.22 mm after fermentation to obtain a fruit and vegetable enzyme liquid; S4, putting pullulan, polyglycerol ricinoleate, ultrapure water and rapeseed oil into a thermostatic stirrer with a power of 1.6 kW, with a constant temperature of 38°C, a constant speed of 300 r / min, a constant stirring time of 12 h, and reacting under constant temperature stirring to obtain a viscous compound; S5, put β-ionone, vanillin acetate, and anhydrous ethanol into a 1.5 kW stirrer, with a stirring temperature of 25°C, a stirring speed of 300 r / min, and a stirring time of 10 min. After dissolving, add the viscous compound prepared by S4, with a stirring temperature of 25°C, a stirring speed of 300 r / min, and a stirring time of 18 min. After sufficient adsorption, put into a vacuum freeze-nano spray dryer with a power of 2.3 kW, with a vacuum degree of -0.05 MPa, a freeze-nano spray drying temperature of -40°C, a freeze-nano spray drying particle size of 100 nm, and a freeze-nano spray drying time of 2.5 h. After freeze-spray drying, flavor-adjusting nanoparticles are obtained; S6. Put the flavor regulating nanoparticles prepared in S5 and the fruit and vegetable enzyme liquid prepared in S3 into a stirrer with a power of 1.5 kW, stir at a temperature of 20°C, a stirring speed of 300 r / min, and a stirring time of 20 min. Mix them evenly to obtain a kale complex enzyme liquid.

[0026] In S2, the mold agent is composed of Streptomyces herbicola, Mucor circinelloides, and Penicillium oxalicum, and the weight ratio is 1.1:0.9:1. The Streptomyces herbicola was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC4.6834, the Mucor circinelloides was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC3.7107, and the Penicillium oxalicum was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC3.15651. The solid-liquid ratio of the mold agent to 5% sterile glucose solution is 1:100 g / mL.

[0027] In S4, the mass fraction of the pullulan in ultrapure water is 8%, and the mass fraction of polyglycerol ricinoleate in rapeseed oil is 6%.

[0028] In S5, the mass fraction of β-ionone in anhydrous ethanol is 1.7%.

[0029] The method for preparing the nutritional supplement composition specifically comprises the following steps: L1. Put chickpeas, pecan kernels and red snapper into a pulping machine with a power of 1.8 kW, the pulping temperature is 85°C, the pulping time is 35 min, the pulping speed is 18000 r / min, and the pulping is performed to obtain a nutrient slurry; L2, put propylene glycol alginate, sorbitan tristearate, sodium hydroxide, ultrapure water, and olive oil into a constant temperature stirrer with a power of 1.6 kW, the constant temperature stirring temperature is 60°C, the constant temperature stirring speed is 300 r / min, the constant temperature stirring time is 6 h, react and form a complex solution; L3. Put the nutrient slurry prepared by L1 and the complex solution prepared by L2 into a high-pressure homogenizer with a power of 1.8 kW, a pressure of 20 MPa, a high-pressure homogenization temperature of 60°C, a high-pressure homogenization time of 12 min, and a high-pressure homogenization speed of 1000 r / min to obtain a nutritional supplement composition.

[0030] This embodiment also provides a method for preparing kale composite fruit and vegetable enzyme, which specifically comprises the following steps: Step 1: Put magnesium sulfate, calcium chloride, zinc sulfate, L-cysteine ​​and ultrapure water into a 1.5 kW stirrer at a stirring temperature of 28°C, a stirring speed of 400 r / min, a stirring time of 12 min, and stir to mix to obtain an active maintenance agent; Step 2: Add the kale complex enzyme solution to the activity maintaining agent prepared in step 1, stir at a temperature of 28° C., a stirring speed of 400 r / min, and a stirring time of 15 min, and mix well to obtain the kale enzyme solution; Step 3: Add the nutritional supplement composition to the kale enzyme solution prepared in step 2, use an ultrasonic agitator with a power of 1.8 kW, an ultrasonic frequency of 15 kHz, a stirring speed of 20,000 r / min, a stirring temperature of 25°C, and a stirring time of 10 min, and obtain the kale composite fruit and vegetable enzyme after uniform dispersion.

[0031] In step 1, the mass fraction of magnesium sulfate in ultrapure water is 1.4%.

[0032] Example 2: This example provides a kale composite fruit and vegetable enzyme and a preparation method thereof. The kale composite fruit and vegetable enzyme comprises the following components in parts by weight: 50 parts of kale complex enzyme liquid, 17 parts of nutritional supplement composition, 1.1 parts of magnesium sulfate, 1.2 parts of calcium chloride, 0.7 parts of zinc sulfate, and 1 part of L-cysteine.

[0033] The raw materials for preparing the kale complex enzyme solution include the following components in parts by weight: 26 parts of kale, 17 parts of grapefruit, 21 parts of prunes, 9 parts of spinach, 8 parts of rocket, 12 parts of Parmesan cheese, 13 parts of quinoa, 1.5 parts of mold agent, 15 parts of pullulan, 18 parts of polyglycerol ricinoleate, 1.2 parts of β-ionone, and 1.1 parts of vanillin acetate.

[0034] The raw materials for preparing the nutritional supplement composition include the following components in parts by weight: 9 parts of chickpeas, 13 parts of pecan kernels, 8 parts of red snapper, 12 parts of propylene glycol alginate, 11 parts of sorbitan tristearate, and 0.9 parts of sodium hydroxide.

[0035] The preparation method of the kale complex enzyme solution specifically comprises the following steps: S1. Put kale, grapefruit, prune, spinach, arugula, Parmesan cheese, and quinoa into a 2.4 kW grinder at a grinding temperature of 23° C., a grinding time of 22 min, and a grinding speed of 3500 r / min to obtain a mixed raw material; S2, placing the mold inoculant and 5% sterile glucose solution in a constant temperature shaking incubator with a power of 2.3 kW, the constant temperature shaking incubation temperature is 28°C, the constant temperature shaking incubation time is 28 h, the constant temperature shaking incubation speed is 230 r / min, and the constant temperature shaking incubation is performed to obtain a mold solution; S3, placing the mixed raw material prepared by S1 and the mold liquid prepared by S2 into a fermentation tank with a power of 2.4 kW, fermenting at a temperature of 31°C, a fermentation speed of 100 r / min, and a fermentation time of 38 days, and filtering with a sterile filter membrane filter with a pore size of 0.22 mm after fermentation to obtain a fruit and vegetable enzyme liquid; S4, putting pullulan, polyglycerol ricinoleate, ultrapure water and rapeseed oil into a thermostatic stirrer with a power of 1.8 kW, with a constant temperature of 42°C, a constant speed of 400 r / min, a constant time of 15 h, and reacting under constant temperature stirring to obtain a viscous compound; S5, put β-ionone, vanillin acetate, and anhydrous ethanol into a stirrer with a power of 1.7 kW, the stirring temperature is 28°C, the stirring speed is 310 r / min, the stirring time is 13 min, and after dissolving, add the viscous compound prepared by S4, the stirring temperature is 30°C, the stirring speed is 300 r / min, the stirring time is 20 min, fully adsorb, put into a vacuum freeze nano spray dryer with a power of 2.6 kW, the vacuum degree is -0.05 MPa, the freeze nano spray drying temperature is -40°C, the freeze nano spray drying particle size is 100 nm, the freeze nano spray drying time is 2.8 h, and freeze spray drying is performed to obtain flavor-adjusting nanoparticles; S6. Put the flavor regulating nanoparticles prepared in S5 and the fruit and vegetable enzyme liquid prepared in S3 into a stirrer with a power of 1.6 kW, stir at a temperature of 23°C, a stirring speed of 330 r / min, and a stirring time of 25 min, mix them evenly to obtain a kale complex enzyme liquid.

[0036] In S2, the mold agent is composed of Streptomyces herbicola, Mucor circinelloides, and Penicillium oxalicum, and the weight ratio is 1.1:0.9:1. The Streptomyces herbicola was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC4.6834, the Mucor circinelloides was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC3.7107, and the Penicillium oxalicum was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC3.15651. The solid-liquid ratio of the mold agent to 5% sterile glucose solution is 1:100 g / mL.

[0037] In S4, the mass fraction of the pullulan in ultrapure water is 10%, and the mass fraction of polyglycerol ricinoleate in rapeseed oil is 7%.

[0038] In S5, the mass fraction of β-ionone in anhydrous ethanol is 2.3%.

[0039] The method for preparing the nutritional supplement composition specifically comprises the following steps: L1. Put chickpeas, pecan kernels and red snapper into a 2.2 kW refiner at a refining temperature of 88°C, a refining time of 37 min and a refining speed of 19000 r / min to obtain a nutrient slurry; L2, put propylene glycol alginate, sorbitan tristearate, sodium hydroxide, ultrapure water, and olive oil into a constant temperature stirrer with a power of 1.7 kW, the constant temperature stirring temperature is 65°C, the constant temperature stirring speed is 400 r / min, the constant temperature stirring time is 7 h, react and form a complex solution; L3. Put the nutrient slurry prepared by L1 and the complex solution prepared by L2 into a high-pressure homogenizer with a power of 1.8 kW, a pressure of 20 MPa, a high-pressure homogenization temperature of 60°C, a high-pressure homogenization time of 13 min, and a high-pressure homogenization speed of 1500 r / min to obtain a nutritional supplement composition.

[0040] This embodiment also provides a method for preparing kale composite fruit and vegetable enzyme, which specifically comprises the following steps: Step 1: Put magnesium sulfate, calcium chloride, zinc sulfate, L-cysteine ​​and ultrapure water into a 1.6 kW stirrer at a stirring temperature of 30°C, a stirring speed of 500 r / min, a stirring time of 15 min, and stir to mix to obtain an active maintenance agent; Step 2: Add the kale complex enzyme solution to the activity maintaining agent prepared in step 1, stir at a temperature of 30° C., a stirring speed of 500 r / min, and a stirring time of 18 min, and mix well to obtain the kale enzyme solution; Step 3: Add the nutritional supplement composition to the kale enzyme solution prepared in step 2, use an ultrasonic agitator with a power of 1.8 kW, an ultrasonic frequency of 15 kHz, a stirring speed of 20,000 r / min, a stirring temperature of 25°C, and a stirring time of 10 min, and obtain the kale composite fruit and vegetable enzyme after uniform dispersion.

[0041] In step 1, the mass fraction of magnesium sulfate in ultrapure water is 1.4%.

[0042] Example 3: This example provides a kale composite fruit and vegetable enzyme and a preparation method thereof. The kale composite fruit and vegetable enzyme comprises the following components in parts by weight: 60 parts of kale complex enzyme liquid, 19 parts of nutritional supplement composition, 1.2 parts of magnesium sulfate, 1.3 parts of calcium chloride, 0.9 parts of zinc sulfate, and 1.2 parts of L-cysteine.

[0043] The raw materials for preparing the kale complex enzyme solution include the following components in parts by weight: 29 parts of kale, 18 parts of grapefruit, 22 parts of prunes, 11 parts of spinach, 10 parts of rocket, 13 parts of Parmesan cheese, 14 parts of quinoa, 1.7 parts of mold agent, 17 parts of pullulan, 21 parts of polyglycerol ricinoleate, 1.4 parts of β-ionone, and 1.3 parts of vanillin acetate.

[0044] The raw materials for preparing the nutritional supplement composition include the following components in parts by weight: 10 parts of chickpeas, 15 parts of pecan kernels, 10 parts of red snapper, 13 parts of propylene glycol alginate, 12 parts of sorbitan tristearate, and 1.1 parts of sodium hydroxide.

[0045] The preparation method of the kale complex enzyme solution specifically comprises the following steps: S1. Put kale, grapefruit, prune, spinach, arugula, Parmesan cheese, and quinoa into a 2.6 kW grinder at a grinding temperature of 26° C., a grinding time of 26 min, and a grinding speed of 4000 r / min to obtain a mixed raw material; S2, placing the mold inoculant and 5% sterile glucose solution in a constant temperature shaking incubator with a power of 2.4 kW, the constant temperature shaking incubation temperature is 28°C, the constant temperature shaking incubation time is 30 h, the constant temperature shaking incubation speed is 240 r / min, and the constant temperature shaking incubation is performed to obtain a mold solution; S3, placing the mixed raw material prepared by S1 and the mold liquid prepared by S2 into a fermentation tank with a power of 2.6 kW, fermenting at a temperature of 33°C, a fermentation speed of 110 r / min, and a fermentation time of 42 days, and filtering with a sterile filter membrane filter with a pore size of 0.22 mm after fermentation to obtain a fruit and vegetable enzyme liquid; S4, putting pullulan, polyglycerol ricinoleate, ultrapure water and rapeseed oil into a thermostatic stirrer with a power of 1.9 kW, with a constant temperature of 55°C, a constant speed of 500 r / min, a constant time of 16 h, and reacting under constant temperature stirring to obtain a viscous compound; S5, put β-ionone, vanillin acetate, and anhydrous ethanol into a stirrer with a power of 1.8 kW, the stirring temperature is 33°C, the stirring speed is 320 r / min, and the stirring time is 15 min. After dissolving, add the viscous compound prepared by S4, the stirring temperature is 33°C, the stirring speed is 320 r / min, and the stirring time is 22 min. After sufficient adsorption, put it into a vacuum freeze nano spray dryer with a power of 2.8 kW, the vacuum degree is -0.05 MPa, the freeze nano spray drying temperature is -40°C, the freeze nano spray drying particle size is 100 nm, the freeze nano spray drying time is 3 h, and freeze spray drying is performed to obtain flavor-adjusting nanoparticles; S6. Put the flavor regulating nanoparticles prepared in S5 and the fruit and vegetable enzyme liquid prepared in S3 into a stirrer with a power of 1.8 kW, stir at a temperature of 26°C, a stirring speed of 400 r / min, and a stirring time of 30 min. Mix them evenly to obtain a kale complex enzyme liquid.

[0046] In S2, the mold agent is composed of Streptomyces herbicola, Mucor circinelloides, and Penicillium oxalicum, and the weight ratio is 1.1:0.9:1. The Streptomyces herbicola was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC4.6834, the Mucor circinelloides was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC3.7107, and the Penicillium oxalicum was purchased from the China General Microbiological Culture Collection Administration Center with a preservation number of CGMCC3.15651. The solid-liquid ratio of the mold agent to 5% sterile glucose solution is 1:100 g / mL.

[0047] In S4, the mass fraction of the pullulan in ultrapure water is 12%, and the mass fraction of polyglycerol ricinoleate in rapeseed oil is 9%.

[0048] In S5, the mass fraction of the β-ionone in the anhydrous ethanol is 2.6%.

[0049] The method for preparing the nutritional supplement composition specifically comprises the following steps: L1. Put chickpeas, pecan kernels and red snapper into a 2.4 kW refiner at a refining temperature of 90°C, a refining time of 40 min and a refining speed of 20,000 r / min to obtain a nutrient slurry; L2, put propylene glycol alginate, sorbitan tristearate, sodium hydroxide, ultrapure water, and olive oil into a constant temperature stirrer with a power of 1.9 kW, the constant temperature stirring temperature is 70°C, the constant temperature stirring speed is 500 r / min, the constant temperature stirring time is 8 h, react and form a complex solution; L3. Put the nutrient slurry prepared by L1 and the complex solution prepared by L2 into a high-pressure homogenizer with a power of 1.8 kW, a pressure of 20 MPa, a high-pressure homogenization temperature of 60°C, a high-pressure homogenization time of 15 min, and a high-pressure homogenization speed of 2000 r / min to obtain a nutritional supplement composition.

[0050] This embodiment also provides a method for preparing kale composite fruit and vegetable enzyme, which specifically comprises the following steps: Step 1: Put magnesium sulfate, calcium chloride, zinc sulfate, L-cysteine ​​and ultrapure water into a stirrer with a power of 1.8 kW, stir at a temperature of 33°C, a stirring speed of 600 r / min, and a stirring time of 18 min, stir and mix to obtain an active maintenance agent; Step 2: Add the kale complex enzyme solution to the activity maintaining agent prepared in step 1, stir at a temperature of 33° C., a stirring speed of 600 r / min, and a stirring time of 20 min, and mix well to obtain the kale enzyme solution; Step 3: Add the nutritional supplement composition to the kale enzyme solution prepared in step 2, use an ultrasonic agitator with a power of 1.8 kW, an ultrasonic frequency of 15 kHz, a stirring speed of 20,000 r / min, a stirring temperature of 25°C, and a stirring time of 10 min, and obtain the kale composite fruit and vegetable enzyme after uniform dispersion.

[0051] In step 1, the mass fraction of magnesium sulfate in ultrapure water is 1.4%.

[0052] Comparative Example 1: This comparative example provides a composite fruit and vegetable enzyme and a preparation method thereof. The only difference from Example 1 is that the added kale composite enzyme liquid does not contain flavor regulating nanoparticles, and the remaining components, component contents, and method steps are the same as those in Example 1.

[0053] Comparative Example 2: This comparative example provides a composite fruit and vegetable enzyme and a preparation method thereof, which is different from Example 1 only in that the added nutritional supplement composition does not contain a complex solution, and the remaining components, component contents, and method steps are the same as those in Example 1.

[0054] Experimental Example 1: Efficacy and stability determination test.

[0055] The test steps for determining the efficacy and stability of the kale composite fruit and vegetable enzyme prepared in Examples 1-3 of the present invention are as follows: (1) Thirty-five Kunming mice weighing 25 g were randomly divided into five groups, with seven mice in each group. The mice in each group were deprived of water but not food for 72 h at a temperature of 24°C and a humidity of 50% to induce a constipation model caused by dryness and dehydration. (2) The mice in Example 1-3 groups were fed 3.5 mL of the kale composite fruit and vegetable enzyme prepared in Example 1-3 every day, the mice in Comparative Example 1-2 groups were fed 3.5 mL of the composite fruit and vegetable enzyme prepared in Comparative Example 1-2 every day, and the mice in the control group were fed 3.5 mL of commercially available common fruit and vegetable enzymes (purchased from Xiamen Zhenhailai Biotechnology Co., Ltd.) every day, and the mice were deprived of water but not food for 10 days; (3) 10 days after modeling, the mice in Example 1-3, Comparative Example 1-2, and Control groups were fed with 2% ink suspension prepared with distilled water by gavage. After each mouse was fed 3.5 mL, the mice were placed in a mouse cage with a clean white filter paper underneath. The time when each mouse first excreted black feces and the total number of black feces excreted by the mouse within 5 hours were observed and recorded. (4) The kale composite fruit and vegetable enzyme prepared in Example 1-3, the composite fruit and vegetable enzyme prepared in Comparative Example 1-2, and the commercially available common fruit and vegetable enzyme in the control group were stored in a refrigerator at 4°C for 2 days to obtain the kale composite fruit and vegetable enzyme of Example 1-3 after storage, the composite fruit and vegetable enzyme of Comparative Example 1-2 after storage, and the commercially available common fruit and vegetable enzyme after storage; (5) 35 Kunming mice weighing 25 g were randomly divided into 5 groups, with 7 mice in each group. The mice were kept at a temperature of 24°C and a humidity of 50% with free access to water and food. (6) The mice in Example 1-3 groups were fed 3.5 mL of the stored kale composite fruit and vegetable enzyme prepared in Example 1-3 every day, the mice in Comparative Example 1-2 groups were fed 3.5 mL of the stored composite fruit and vegetable enzyme prepared in Comparative Example 1-2 every day, and the mice in the control group were fed 3.5 mL of the stored commercially available common fruit and vegetable enzyme (purchased from Xiamen Zhenhailai Biotechnology Co., Ltd.) every day. After 10 days of feeding, the mice in Example 1-3 groups, Comparative Example 1-2 groups, and the control group were fed 3.5 mL of a 2% carbon ink suspension prepared with distilled water by gavage. Each mouse was fed 30 minutes after gavage. The mice were killed by cervical dislocation, and the stomach and intestine were immediately removed by laparotomy and spread on a glass plate. The moving distance of the carbon tip in the intestine and the total length of the small intestine (from pylorus to ileum) were measured, and the small intestine advancement percentage was calculated according to the formula: advancement percentage (%) = distance between the front end of the carbon tip and the pylorus / total length of the small intestine × 100%.

[0056] Result analysis: Figure 1 This is a scanning electron microscope image of the internal structure of the viscous compound of the present invention. Figure 2 This is a graph showing the first black discharge time of each group of mice in Experimental Example 1 of the present invention. Figure 3The total number of black feces discharged within 5 hours in each group of mice described in Experimental Example 1 of the present invention is shown in the figure. By gavage, the mice in the dryness and dehydration constipation model groups of Examples 1-3 were fed 3.5 mL of the kale compound fruit and vegetable enzyme prepared in Examples 1-3 every day, and were fed without water or fasting for 10 d. They were gavaged with a 2% ink suspension made of distilled water. The time when the mice in the Example 1-3 groups first discharged black feces was 170.1 min, 164.9 min, and 158.8 min, respectively. The total number of black feces discharged within 5 hours in the Example 1-3 groups of mice was 5.8, 6.7, and 7.3, respectively. By gavage, the mice in the dryness and dehydration constipation model groups of Comparative Examples 1-2 were fed 3.5 mL of the compound fruit and vegetable enzyme prepared in Comparative Examples 1-2 every day, and were fed without water or fasting for 10 d. They were gavaged with a 2% ink suspension made of distilled water. The time when the mice in the Comparative Examples 1-2 groups first discharged black feces was 171.8 min, 179.1 min, the total number of black feces discharged by mice in groups 1-2 of the control group within 5 hours was 5.2 and 4.3 respectively. The mice in the control group with constipation model were fed with 3.5 mL of commercially available common fruit and vegetable enzymes for 10 days without water or fasting by gavage, and 2% ink suspension prepared with distilled water was gavaged. The time for the first discharge of black feces by mice in the control group was 218.6 min, and the total number of black feces discharged by mice in the control group within 5 hours was 2.8; Figure 4 This is a graph showing the intestinal propulsion rate of mice described in Experimental Example 1 of the present invention. As shown in the figure, the mice in groups 1-3 of Examples 1-3 were gavaged with 3.5 mL of the kale composite fruit and vegetable enzyme prepared in Example 1-3 after storage every day. After 10 days, they were gavaged with a 2% carbon ink suspension prepared with distilled water, and the intestinal propulsion rates of the mice in groups 1-3 of Examples 1-3 were 96.3%, 97.0%, and 97.6%, respectively. The mice in groups 1-2 of Comparative Examples 1-2 were gavaged with 3.5 mL of the composite fruit and vegetable enzyme prepared in Example 1-2 after storage every day. After 10 days, they were gavaged with a 2% carbon ink suspension prepared with distilled water, and the intestinal propulsion rates of the mice in groups 1-2 of Comparative Examples 1-2 were 95.8% and 92.6%, respectively. The mice in the control group were gavaged with 3.5 mL of commercially available ordinary fruit and vegetable enzymes after storage every day, and 10 days later, they were gavaged with a 2% carbon ink suspension prepared with distilled water, and the intestinal propulsion rates of the mice in groups 1-2 of Comparative Examples 1-2 were 95.8% and 92.6%, respectively. d later, the mice in the control group were gavaged with a 2% carbon ink suspension prepared with distilled water, and the intestinal propulsion rate was 74.2%, indicating that the kale composite fruit and vegetable enzyme prepared by the present invention can significantly relieve constipation, promote intestinal peristalsis, has stable efficacy, and is not easy to lose activity during storage.

[0057] Experimental Example 2: Nutritional supplementation measurement test.

[0058] The test steps for determining the nutritional supplement of kale composite fruit and vegetable enzymes prepared in Examples 1-3 of the present invention are as follows: (1) The determination method of protein content in kale composite fruit and vegetable enzymes refers to the standard GB 5009.5-2016, the determination method of vitamin B12 content refers to the standard GB 5413.14-2010, the determination method of iron, calcium and zinc refers to the standard GB5413.21-2010, and the determination method of fatty acids refers to the standard GB5009.168-2016; (2) 1 L of the kale composite fruit and vegetable enzyme prepared in Example 1-3 was taken as Example 1-3 group, 1 L of the composite fruit and vegetable enzyme prepared in Comparative Example 1-2 was taken as Comparative Example 1-2 group, and 1 L of commercially available ordinary fruit and vegetable enzyme (purchased from Xiamen Zhenhailai Biotechnology Co., Ltd.) was taken as the control group. Five parallel measurements were set for each group, and the results were recorded in Table 1.

[0059] Table 1. Fruit and vegetable enzyme nutritional components test table

[0060] Result analysis: As shown in Table 1, the protein, vitamin B12, iron, calcium, zinc, and fatty acids in the kale composite fruit and vegetable enzyme prepared in Examples 1-3 are higher than those in the commercially available ordinary fruit and vegetable enzyme in the control group, indicating that the complex solution formed by the carboxyl group of propylene glycol alginate and sorbitan tristearate can form a stable interface layer on the surface of the nutrient slurry to prevent it from direct contact with the external environment, thereby effectively supplementing the nutritional components in the kale composite fruit and vegetable enzyme.

[0061] Experimental Example 3: Flavor and taste measurement test.

[0062] The test steps for determining the flavor and taste of the kale composite fruit and vegetable enzyme prepared in Examples 1-3 of the present invention are as follows: (1) Take 100 mL of the kale composite fruit and vegetable enzyme prepared in Examples 1-3 as the fruit and vegetable enzyme samples of Examples 1-3, take 100 mL of the composite fruit and vegetable enzyme prepared in Comparative Examples 1-2 as the fruit and vegetable enzyme samples of Comparative Examples 1-2, and take 100 mL of commercially available common fruit and vegetable enzyme (purchased from Xiamen Zhenhailai Biotechnology Co., Ltd.) as the fruit and vegetable enzyme sample of the control group, and take 7 parallel samples in each group; (2) Table 2 was used as the sensory test scoring standard for each group of fruit and vegetable enzyme samples. The color, appearance, smell and taste of each group of fruit and vegetable enzyme samples were evaluated and scored, and the average value was recorded. The results are shown in Table 3; Table 2. Fruit and vegetable enzyme sample scoring standard table

[0063] Table 3. Fruit and vegetable enzyme sample scoring table

[0064] Result analysis: Figure 5 This is a finished product of the kale composite fruit and vegetable enzyme of the present invention, as shown in Figure 5 As shown in Table 3, the color, appearance, smell and taste scores of the kale composite fruit and vegetable enzymes prepared in Examples 1-3 are higher than those of the composite fruit and vegetable enzymes prepared in Comparative Example 1 and the commercially available ordinary fruit and vegetable enzymes in the control group, and the color, appearance, smell and taste of the kale composite fruit and vegetable enzymes prepared in Examples 1-3 are stable, indicating that pullulan and polyglycerol ricinoleate are cross-linked through hydrogen bonds to form a network structure, and the obtained viscous compound fully absorbs β-ionone and vanillin acetate with aromatic and sweet odors, thereby improving the smoothness and softness of the kale composite fruit and vegetable enzyme and giving it excellent flavor.

[0065] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

[0066] The present invention and its implementation methods are described above, which is not restrictive. The drawings are only one of the implementation methods of the present invention, and the actual application is not limited thereto. In short, if ordinary technicians in the field are inspired by it and design methods and embodiments similar to the technical solution without creativity without departing from the purpose of the invention, they should all fall within the protection scope of the present invention.

Claims

1. A kale composite fruit and vegetable enzyme, characterized in that: The kale composite fruit and vegetable enzyme comprises the following components: 40-60 parts of kale complex enzyme liquid, 15-19 parts of nutritional supplement composition, 0.8-1.2 parts of magnesium sulfate, 1.1-1.3 parts of calcium chloride, 0.6-0.9 parts of zinc sulfate, and 0.8-1.2 parts of L-cysteine; The raw materials for preparing the kale complex enzyme solution include the following components in parts by weight: 23-29 parts of kale, 16-18 parts of grapefruit, 19-22 parts of prunes, 7-11 parts of spinach, 6-10 parts of rocket, 11-13 parts of Parmesan cheese, 12-14 parts of quinoa, 1.4-1.7 parts of mold agent, 14-17 parts of pullulan, 15-21 parts of polyglycerol ricinoleate, 0.8-1.4 parts of β-ionone, and 0.9-1.3 parts of vanillin acetate; The raw materials for preparing the nutritional supplement composition include the following components in parts by weight: 8-10 parts of chickpeas, 12-15 parts of pecan kernels, 6-10 parts of red snapper, 11-13 parts of propylene glycol alginate, 10-12 parts of sorbitan tristearate, and 0.7-1.1 parts of sodium hydroxide.

2. The kale composite fruit and vegetable enzyme according to claim 1, characterized in that: The preparation method of the kale complex enzyme solution specifically comprises the following steps: S1, crushing kale, grapefruit, prune, spinach, rocket, Parmesan cheese, and quinoa to obtain a mixed raw material; S2, mixing the mold agent with 5% sterile glucose solution, adding the mixed raw material prepared in S1, and fermenting, and filtering with a sterile filter membrane with a pore size of 0.22 mm after fermentation to obtain a fruit and vegetable enzyme liquid; S3, stirring and reacting pullulan with polyglycerol ricinoleate, ultrapure water and rapeseed oil to obtain a viscous compound; S4, dissolving β-ionone and vanillin acetate in anhydrous ethanol, adding the viscous compound prepared in S3, fully adsorbing, and freeze-spray drying to obtain flavor-adjusting nanoparticles; S5. Evenly mix the flavor regulating nanoparticles prepared in S4 and the fruit and vegetable enzyme solution prepared in S2 to obtain kale composite enzyme solution.

3. The kale composite fruit and vegetable enzyme according to claim 2, characterized in that: In S2, the fungal agent consists of Streptomyces herbicola, Mucor circinelloides, and Penicillium oxalicum, with a weight ratio of 1.1:0.9:1, and the solid-liquid ratio of the fungal agent to 5% sterile glucose solution is 1:100 g / mL.

4. The kale composite fruit and vegetable enzyme according to claim 3, characterized in that: In S3, the mass fraction of the pullulan in ultrapure water is 8-12%, and the mass fraction of polyglycerol ricinoleate in rapeseed oil is 6-9%.

5. The kale composite fruit and vegetable enzyme according to claim 4, characterized in that: In S4, the mass fraction of the β-ionone in the anhydrous ethanol is 1.7-2.6%.

6. The kale composite fruit and vegetable enzyme according to claim 5, characterized in that: The method for preparing the nutritional supplement composition specifically comprises the following steps: L1, mixing chickpeas, pecan kernels and red snapper, and grinding them to obtain a nutrient slurry; L2, stirring propylene glycol alginate, sorbitan tristearate, sodium hydroxide, ultrapure water, and olive oil to react to form a complex solution; L3. The nutrient slurry prepared in L1 and the complex solution prepared in L2 are homogenized under high pressure to obtain a nutritional supplement composition.

7. The kale composite fruit and vegetable enzyme according to claim 6, characterized in that: In L2, the mass fraction of propylene glycol alginate in ultrapure water is 5-8%, and the mass fraction of sorbitan tristearate in olive oil is 4-6%.

8. A method for preparing the kale composite fruit and vegetable enzyme according to any one of claims 1 to 7, characterized in that: The specific steps include: Step 1: Stir and fully dissolve magnesium sulfate, calcium chloride, zinc sulfate, L-cysteine, and ultrapure water to obtain an activity maintaining agent; Step 2, adding the kale complex enzyme solution to the activity maintaining agent prepared in step 1, and mixing to obtain the kale enzyme solution; Step 3: Evenly disperse the nutritional supplement composition in the kale enzyme solution prepared in step 2 to obtain kale composite fruit and vegetable enzyme.

9. The method for preparing the kale composite fruit and vegetable enzyme according to claim 8, characterized in that: In step 1, the mass fraction of magnesium sulfate in ultrapure water is 1.4%.

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