CN113040305A - Fermented pumpkin pulp rich in pumpkin polysaccharide and preparation method thereof - Google Patents
Fermented pumpkin pulp rich in pumpkin polysaccharide and preparation method thereof Download PDFInfo
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- 235000015136 pumpkin Nutrition 0.000 title claims abstract description 201
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- 229960001790 sodium citrate Drugs 0.000 claims description 9
- 235000010352 sodium erythorbate Nutrition 0.000 claims description 9
- 239000004320 sodium erythorbate Substances 0.000 claims description 9
- RBWSWDPRDBEWCR-RKJRWTFHSA-N sodium;(2r)-2-[(2r)-3,4-dihydroxy-5-oxo-2h-furan-2-yl]-2-hydroxyethanolate Chemical compound [Na+].[O-]C[C@@H](O)[C@H]1OC(=O)C(O)=C1O RBWSWDPRDBEWCR-RKJRWTFHSA-N 0.000 claims description 9
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- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241000408747 Lepomis gibbosus Species 0.000 description 1
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- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
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- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 238000009924 canning Methods 0.000 description 1
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- 230000002349 favourable effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
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- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
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- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Preparation Of Fruits And Vegetables (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly relates to a preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide, and the fermented pumpkin pulp rich in pumpkin polysaccharide. According to the invention, firstly, the pumpkin pulp is treated by using the ultrasonic-assisted compound enzyme method to fully release pumpkin polysaccharide, then the growth promoter for promoting probiotics to produce polysaccharide is added, and then the polysaccharide is further produced in a fermentation mode, so that the polysaccharide content in the fermented pumpkin pulp can be greatly improved, and compared with the polysaccharide content in the traditional fermentation method, the polysaccharide content in the fermented pumpkin pulp prepared by the method is improved by about 1 time. Repeated experiments prove that the growth and reproduction capacity of probiotics in the pumpkin pulp can be improved by adding dipotassium phosphate, manganese sulfate and the like as growth promoters, and the polysaccharide content of the pumpkin pulp is improved by utilizing the fermentation of the probiotics to generate more polysaccharide substances, so that the polysaccharide content in the product can reach above 156.43 mg/L.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide, and the fermented pumpkin pulp rich in pumpkin polysaccharide.
Background
Pumpkin polysaccharide is a sugar chain formed by bonding many different types of glycosidic bonds, and mainly consists of rhamnose, arabinose, glucose and galactose. A large number of researches show that the pumpkin polysaccharide has the functions of resisting oxidation, reducing the blood sugar concentration, inhibiting tumor bodies, improving the immunity of organisms and the like. Currently, research on pumpkins is more, for example:
the pumpkin preparation method disclosed in the Chinese invention 'pumpkin whole pulp and preparation method' with publication number CN108056422A improves the utilization rate of pumpkin through the treatments of pumpkin cleaning, cutting, crushing, pulp boiling, pulping, slag removal, homogenization and the like, simplifies the processing steps, and the pumpkin whole pulp with the soluble solid content of 6-15 percent, the insoluble solid content of 25-50 percent and the pH value of 5.8-6.8 is processed. The preparation method comprises the following steps: A. cleaning: cleaning the pumpkin; B. cutting into sections; C. crushing: crushing and adding softened water to obtain pumpkin pulp; D. boiling the soybean milk: heating the pulp to 95-100 deg.C, and maintaining for 5-10 min; E. pulping: pulping and refining the softened pumpkin pulp; f: cooling; g: deslagging: treating the pumpkin pulp by a horizontal slag remover to remove hard pumpkin seed shells; h: homogenizing; i: sterilizing; j: canning and storing. The invention maximally utilizes the pumpkin, refines the pumpkin whole pulp through double pulping, deslagging and homogenizing processes, and obtains the pumpkin whole pulp product with rich nutrition, bright color value, smooth taste and good flavor.
Publication No.: CN105962196A, the Chinese invention "method for fermenting pumpkin pulp by plant probiotics" discloses a method for fermenting pumpkin pulp by plant probiotics, which adopts two enzymolysis processes, uses high-temperature amylase for one-time enzymolysis, can effectively reduce the viscosity of pumpkin, and improves the pulp yield to about 95%; the secondary enzymolysis uses medium-temperature amylase and glucoamylase, so that the sweet feeling of the pumpkin can be effectively improved, the viscosity is reduced again, the pumpkin is endowed with better aroma and fine and semitransparent texture, and the taste of the pumpkin is fuller and sweeter. After the two times of enzymolysis, the soluble solid of the pumpkin is improved by 1 to 2 points, thereby reducing about 30 percent of the addition amount of glucose. The method of the invention properly reduces the viscosity of the pumpkin puree through an enzymolysis process and a plant probiotic fermentation process, simultaneously generates lactic acid and salt substances thereof with fresh-keeping function, reduces the pH value of the product, effectively reduces the sterilization condition of the pumpkin puree, expands the application width of the pumpkin product in the food industry and prolongs the quality guarantee period of the raw materials of the pumpkin product.
The invention of China with publication No. CN110419700A, a preparation method of fermented pumpkin powder with high probiotic content and low sugar, discloses a preparation method of fermented pumpkin powder with high probiotic content and low sugar, which comprises the steps of inoculating bacillus coagulans, lactobacillus and yeast as composite fermentation strains to pumpkin slurry prepared after cleaning and seed removing treatment through the action of various probiotics and enzymolysis, performing co-fermentation of the composite strains and enzymolysis of an enzyme preparation to obtain pumpkin slurry (powder) with low sugar content and rich viable bacteria, wherein sugar in the product can be fully utilized by the viable bacteria for growth of the probiotic bacteria, and finally reducing the reduced sugar content of the pumpkin powder to below 5 percent, and the total number of the probiotic bacteria can reach 106cfu/g to 108cfu/g, not only has higher bacteria count but also has low sugar content, and is convenient for diabetes patients and other patients needing blood sugar control.
The normal-temperature live bacterium beverage disclosed in the invention CN107198076A of China 'an Probiotics Normal-temperature live bacterium beverage of Bacillus coagulans and a preparation method thereof' is prepared by carrying out enzymolysis on pumpkin pulp through pectinase, cellulase and carbohydrase, mixing with the Bacillus coagulans, fermenting, adding a flavoring agent, and blending, wherein the pumpkin pulp is prepared by boiling pumpkin blocks and then adding water for pulping; the pumpkin cake seasoning comprises the following raw materials in parts by weight, wherein per 100 parts by weight of pumpkin block raw materials are sequentially added with pectinase, cellulase, glucoamylase and a seasoning in parts by weight: 0.18-0.5 part of pectinase, 0.5-1.8 parts of cellulase, 0.25-1.5 parts of saccharifying enzyme and 4.75-7.25 parts of flavoring agent; the bacillus coagulans probiotic normal-temperature live bacterium beverage prepared by the preparation method disclosed by the invention not only maintains the unique fragrance of pumpkin, but also has strong fermentation fragrance, is moderate in sour and sweet, is coordinated in flavor, contains probiotic bacillus coagulans which is easy to store at normal temperature, and is easy to store at normal temperature.
The method for extracting pumpkin polysaccharide by using the ultrasonic compound enzyme method is common, but organic solvents such as ethanol and the like are added in the conventional common extraction method, so that the pumpkin polysaccharide is not suitable for direct eating by people. The method has mild extraction conditions, does not add organic solvents, can maintain other nutrient components in the extracted pumpkin pulp not to be damaged, and can further perform probiotic fermentation on the reaction liquid to improve the polysaccharide content in the pumpkin pulp.
Disclosure of Invention
The invention firstly utilizes an ultrasonic-assisted compound enzyme method to treat pumpkin pulp to fully release pumpkin polysaccharide, and then adds a growth promoter for promoting probiotics to produce polysaccharide to further produce polysaccharide in a fermentation mode. The invention aims to provide a preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide, which comprises the following steps:
1) peeling and removing seeds of pumpkin, cutting the pumpkin into uniform slices, blanching, adding purified water, crushing and pulping to obtain crushed pulp, and adding sodium erythorbate in the crushing and pulping process;
2) mixing the crushed pulp in the step 1) with a compound enzyme beneficial to dissolving out pumpkin polysaccharide, and performing ultrasonic-assisted enzymolysis to obtain enzymolysis pumpkin pulp; the complex enzyme comprises pectinase, cellulase and papain; the adding mass of the complex enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed slurry respectively;
3) adding glucose, dipotassium phosphate, sodium citrate, manganese sulfate and yeast extract into the enzymatic hydrolysis pumpkin pulp obtained in the step 2), uniformly mixing, and homogenizing, wherein the substances are all probiotic growth promoters; the adding mass of each growth promoting agent is respectively 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymolysis pumpkin pulp;
4) and (3) sterilization: sterilizing for 5-10min under the sterilization condition of 100-110 ℃ to obtain sterilized pumpkin pulp;
5) inoculating compound probiotic strains into the sterilized pumpkin pulp obtained in the step 4) for fermentation for 20-28 h; the composite probiotic strain consists of lactobacillus plantarum and streptococcus thermophilus; the strain proportion of the composite probiotics is 1: 1.
preferably, the addition amount of the sodium erythorbate is 2-4 per mill of the mass of the pumpkin pulp.
Preferably, the ultrasonic-assisted enzymolysis sequentially comprises an ultrasonic treatment process and an enzymolysis process; the time of the ultrasonic treatment process is 10-15 min, and the ultrasonic intensity of the ultrasonic treatment is 4-5W/mL.
Preferably, the temperature of the enzymolysis process is 50-55 ℃, and the time of the enzymolysis process is 40-100 min.
Preferably, the independent enzyme activities of the pectinase, the cellulase and the papain in the compound enzyme are 15-25 ten thousand U/g.
Preferably, the inoculation concentration of the composite probiotics in the step 5) is 107～108CFU/mL, the inoculation amount is 6-8% of the mass of the pumpkin pulp.
Preferably, the fermentation is finished when the pH value of the fermentation liquid in the step 5) is 3.5-4.0.
The invention also provides the fermented pumpkin pulp rich in pumpkin polysaccharide, which is prepared by the preparation method.
Advantageous effects
The invention provides a preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide. The pumpkin polysaccharide is released by adopting an ultrasonic compound enzyme method, after the compound enzyme which is favorable for dissolving out the pumpkin polysaccharide is added for enzymolysis, ultrasonic-assisted enzymolysis is carried out, and the pumpkin polysaccharide is fully dissolved and released through an ultrasonic cavitation effect, so that the yield of the pumpkin polysaccharide is improved, and the polysaccharide content of the raw material is improved as much as possible; then, a fermentation mode is adopted, and the composite probiotic growth promoter is added, so that the growth of composite probiotics in the pumpkin pulp is facilitated, the pumpkin polysaccharide is further released and generated through fermentation, and meanwhile, probiotic metabolites such as short-chain fatty acid, polypeptide and the like can be generated, so that the nutrition of the pumpkin pulp product is comprehensive and balanced. Repeated experiments prove that the growth and reproduction capacity of probiotics in the pumpkin pulp can be improved by adding dipotassium phosphate, manganese sulfate and the like as growth promoters, more polysaccharide substances are generated by fermentation of the probiotics, the polysaccharide content of the pumpkin pulp is improved, and the polysaccharide content in the product can reach above 156.43 mg/L. The combination of the two process methods is adopted to finally improve the pumpkin polysaccharide content in the pumpkin pulp, so that the probiotic fermented pumpkin pulp drink rich in pumpkin polysaccharide is produced, and meanwhile, the pumpkin pulp has more comprehensive and balanced nutrition due to the generation of probiotic metabolites such as short-chain fatty acid, polypeptide and the like.
The invention has simple process technology, abundant and easily obtained raw materials, and is suitable for implementation and popularization.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The lactic acid bacteria adopted by the embodiments of the invention come from the industrial microorganism culture collection and management center (CICC), and the strains are as follows: lactobacillus plantarum subspecies plantaris (CICC21824), Streptococcus thermophilus (CICC20373), are dominant strains suitable for fermentation of pumpkin pulp after strain selection.
Pectinase and cellulase were purchased from Ningxia and Debi biotechnology, Inc., and papain was purchased from Nanning Pompe bioengineering, Inc.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
A preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide comprises the following steps:
1) slicing and blanching: washing fresh and non-rotten pumpkins for 2-3 times by flowing clear water, removing seeds and peels, cutting the pumpkins into uniform slices of 5mm, and blanching the pumpkins in boiling water for 5 min;
2) crushing and pulping: adding purified water with the same volume into the blanched pumpkin slices, adding sodium erythorbate with the total volume of 2 per mill, and pulping to prepare pumpkin pulp;
3) ultrasonic wave and enzyme treatment: adding a complex enzyme preparation with strong enzymolysis capability and beneficial to dissolving out pumpkin polysaccharide into the pumpkin pulp, placing the pumpkin pulp in an ultrasonic crusher for ultrasonic treatment for 10min, and after the ultrasonic treatment is finished, continuing carrying out water bath oscillation enzymolysis for 80min at 55 ℃. The compound enzyme comprises pectinase, cellulase and papain, the adding mass of each enzyme in the compound enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed pulp respectively, and the ultrasonic intensity is 4W/mL;
the independent enzyme activities of the pectinase, the cellulase and the papain in the compound enzyme are respectively 25 ten thousand U/g, 15 ten thousand U/g and 10 ten thousand U/g;
4) adding a probiotic growth promoter: adding glucose, dipotassium hydrogen phosphate, sodium citrate, manganese sulfate and yeast extract into the enzymatic hydrolysis pumpkin pulp, wherein the adding mass is 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymatic hydrolysis pumpkin pulp respectively;
5) homogenizing: homogenizing the pumpkin pulp added with the probiotic growth promoter under the homogenizing pressure of 20MPa and at the homogenizing temperature of 45 ℃;
6) and (3) sterilization: sterilizing the homogenized pumpkin pulp at 100 deg.C for 10 min;
7) inoculation: sterilizing the pumpkin pulp, quickly cooling to 34 ℃, and mixing the composite probiotic agent according to the proportion of 108Inoculating CFU/mL into the pumpkin pulp, wherein the inoculation amount is 8%, fermenting for 20h at 34 ℃, and finishing fermentation when the fermentation pH value is 4.0;
the composite probiotics comprise lactobacillus plantarum and streptococcus thermophilus; the ratio of the strains is 1: 1;
8) the fermented pumpkin pulp prepared by the method has the pumpkin polysaccharide content of 156.43mg/L through detection.
Example 2
A preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide comprises the following steps:
1) slicing and blanching: washing fresh non-rotten pumpkins for 2-3 times by flowing clear water, removing seeds and peels, cutting the pumpkins into uniform slices of 5mm, and blanching the pumpkins in boiling water for 4 min;
2) crushing and pulping: adding purified water with the same volume into the blanched pumpkin slices, adding sodium erythorbate with the total volume of 4 per mill, and pulping to prepare pumpkin pulp;
3) ultrasonic wave and enzyme treatment: adding a compound enzyme preparation with strong enzymolysis capability and beneficial to dissolving out pumpkin polysaccharide into the pumpkin pulp, placing the pumpkin pulp in an ultrasonic crusher for ultrasonic treatment for 15min, and after the ultrasonic treatment is finished, continuing carrying out water bath oscillation enzymolysis for 100min at 50 ℃. The compound enzyme comprises pectinase, cellulase and papain, the adding mass of each enzyme of the compound enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed pulp, and the ultrasonic intensity is 4.5W/mL;
the independent enzyme activities of the pectinase, the cellulase and the papain in the compound enzyme are respectively 20 ten thousand U/g, 25 ten thousand U/g and 15 ten thousand U/g;
4) adding a probiotic growth promoter: adding glucose, dipotassium hydrogen phosphate, sodium citrate, manganese sulfate and yeast extract agent into the enzymatic hydrolysis pumpkin pulp, wherein the adding mass is respectively 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymatic hydrolysis pumpkin pulp;
5) homogenizing: homogenizing the pumpkin pulp added with the probiotic growth promoter under the homogenizing pressure of 25MPa and at the homogenizing temperature of 40 ℃;
6) and (3) sterilization: sterilizing the homogenized pumpkin pulp at 105 deg.C for 6 min;
7) inoculation: sterilizing the pumpkin pulp, quickly cooling to 30 ℃, and mixing the composite probiotic agent according to the proportion of 107Inoculating CFU/mL into the pumpkin pulp, wherein the inoculation amount is 8%, fermenting for 24h at 34 ℃, and finishing fermentation when the fermentation pH value is 4.0;
the composite probiotics comprise lactobacillus plantarum and streptococcus thermophilus; the ratio of the strains is 1: 1.
8) The fermented pumpkin pulp prepared by the method has the pumpkin polysaccharide content of 158.56mg/L through detection.
Example 3
A preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide comprises the following steps:
1) slicing and blanching: washing fresh non-rotten pumpkins for 2-3 times by flowing clear water, removing seeds and peels, cutting the pumpkins into uniform slices of 5mm, and blanching the pumpkins in boiling water for 5 min;
2) crushing and pulping: adding purified water with the same volume into the blanched pumpkin slices, adding sodium erythorbate with the total volume of 4 per mill, and pulping to prepare pumpkin pulp;
3) ultrasonic wave and enzyme treatment: adding a complex enzyme preparation with strong enzymolysis capability and beneficial to dissolving out pumpkin polysaccharide into the pumpkin pulp, placing the pumpkin pulp in an ultrasonic crusher for 12min, and after the ultrasonic treatment, continuing to carry out water bath oscillation enzymolysis for 40min at 55 ℃. The compound enzyme comprises pectinase, cellulase and papain, the adding mass of each enzyme of the compound enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed pulp respectively, and the ultrasonic intensity is 5W/mL;
the independent enzyme activities of the pectinase, the cellulase and the papain in the compound enzyme are respectively 10 ten thousand U/g, 15 ten thousand U/g and 10 ten thousand U/g;
4) adding a probiotic growth promoter: adding glucose, dipotassium hydrogen phosphate, sodium citrate, manganese sulfate and yeast extract into the enzymatic hydrolysis pumpkin pulp, wherein the adding mass is 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymatic hydrolysis pumpkin pulp respectively;
5) homogenizing: homogenizing the pumpkin pulp added with the probiotic growth promoter under the homogenizing pressure of 22MPa and at the homogenizing temperature of 43 ℃;
6) and (3) sterilization: sterilizing the homogenized pumpkin pulp at 103 deg.C for 8 min;
7) inoculation: sterilizing the pumpkin pulp, quickly cooling to 32 ℃, and mixing the composite probiotic agent according to the proportion of 108CFU/mL is inoculated in the pumpkin pulp,the inoculation amount is 6%, the fermentation is carried out for 20h at 32 ℃, and the fermentation is finished when the fermentation pH value is 3.8;
the composite probiotics comprise lactobacillus plantarum and streptococcus thermophilus; the ratio of the strains is 1: 1;
8) the fermented pumpkin pulp prepared by the method has the pumpkin polysaccharide content of 160.23mg/L through detection.
Control test:
a control example was set up to illustrate the effect of different preparation methods on the polysaccharide content of pumpkin pulp. Comparative example 1: the comparative example 1 is different from the examples 1 to 3 in that: the ultrasonic-assisted pectinase-cellulase-papain complex enzyme treatment is not carried out on the pumpkin pulp, and other process steps are the same as those of the invention. The pumpkin pulp obtained by final fermentation has the pumpkin polysaccharide content of 104.78 mg/L. Comparative example 2: the comparative example is different from examples 1 to 3 in that: the probiotic growth promoter glucose, dipotassium phosphate, sodium citrate, manganese sulfate and yeast extract are not added into the pumpkin pulp, and other process steps are the same as those of the invention. The pumpkin pulp obtained by final fermentation has the pumpkin polysaccharide content of 112.42 mg/L. Comparative example 3: the comparative example is different from examples 1 to 3 in that: 1. the pumpkin pulp is not subjected to ultrasonic-assisted pectinase-cellulase-papain complex enzyme treatment; 2. the probiotic growth promoter glucose, dipotassium phosphate, sodium citrate, manganese sulfate and yeast extract are not added into the pumpkin pulp, and other process steps are the same as those of the invention. The pumpkin pulp obtained by final fermentation has the pumpkin polysaccharide content of 81.62 mg/L.
The pumpkin polysaccharide is determined by the following method:
1. the experimental method comprises the following steps:
extracting total polysaccharide by water extraction and alcohol precipitation method, and measuring the content of the total polysaccharide by phenol-sulfuric acid method.
The determination step comprises:
2. the extraction method comprises the following steps:
5mL of the pumpkin slurry of the invention and each control example are respectively sucked, 1mL of deionized water is added, and the mixture is fully homogenized. Extracting in 100 deg.C water bath for 2h (cover must be closed to prevent water loss), cooling, centrifuging at 10000g for 10min, and collecting supernatant. Sucking 0.2mL of supernatant, slowly adding 0.8mL of absolute ethyl alcohol, uniformly mixing, standing overnight at 4 ℃, centrifuging 10min at 10000g, removing supernatant, adding 1mL of water into precipitate, and fully and uniformly mixing to dissolve the precipitate.
3. The determination step comprises:
the microplate reader wavelength was adjusted to 490 nm. Sucking 200 μ L of the supernatant of the treatment solution, adding 100 μ L of reagent I and 0.5mL of concentrated sulfuric acid, mixing, performing water bath at 90 ℃ for 20min, and cooling with running water. 200 mu L of supernatant was added to an ELISA plate, and the absorbance A was measured at 490 nm.
4. And (3) calculating the content:
glucose was used as a control and the regression equation was determined under standard conditions as y-7.981 x-0.0037, R2When the concentration was 0.9973, x was the glucose content (mg/mL) and y was the absorbance.
Total polysaccharide content (μ g/g dry weight) — (a +0.0037) ÷ 7.981 × V1÷V2×V3÷W×1000
＝626.49×(A+0.0037)÷W
V1: the volume of the re-dissolved solution after alcohol precipitation is 1 mL; v2: the volume of alcohol precipitation is 0.2 mL; v3Adding 1mL of water during extraction; w: sample mass, g; 1000, mg to μ g.
5. The measurement data are shown in the following table:
the content of the polysaccharide in the same pumpkin sample produced by adopting different processes is shown in the table:
finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, those skilled in the art will appreciate that; the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. A preparation method of fermented pumpkin pulp rich in pumpkin polysaccharide comprises the following steps:
1) peeling and removing seeds of pumpkin, cutting the pumpkin into uniform slices, blanching, adding purified water, crushing and pulping to obtain crushed pulp, and adding sodium erythorbate in the crushing and pulping process;
2) mixing the crushed pulp in the step 1) with a compound enzyme beneficial to dissolving out pumpkin polysaccharide, and performing ultrasonic-assisted enzymolysis to obtain enzymolysis pumpkin pulp; the complex enzyme comprises pectinase, cellulase and papain; the adding mass of the complex enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed slurry respectively;
3) adding glucose, dipotassium phosphate, sodium citrate, manganese sulfate and yeast extract into the enzymatic hydrolysis pumpkin pulp obtained in the step 2), uniformly mixing and homogenizing; the adding mass is respectively 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymolysis pumpkin pulp;
4) and (3) sterilization: sterilizing the pumpkin pulp at 100-110 ℃ for 5-10 min;
5) inoculating compound probiotics into the sterilized pumpkin pulp obtained in the step 4) for fermentation for 20-28 h; the composite probiotics comprise lactobacillus plantarum and streptococcus thermophilus; the ratio of the strains is 1: 1.
2. The preparation method according to claim 1, wherein the sodium erythorbate is added in an amount of 2 to 4% by mass of the pumpkin pulp.
3. The preparation method according to claim 1, wherein the ultrasonic-assisted enzymolysis sequentially comprises an ultrasonic treatment process and an enzymolysis process; the time of the ultrasonic treatment process is 10-15 min, and the ultrasonic intensity is 4-5W/mL.
4. The preparation method according to claim 3, wherein the temperature of the enzymolysis process is 50-55 ℃ and the time is 40-100 min.
5. The preparation method of claim 1, wherein the independent enzyme activities of the pectinase, the cellulase and the papain in the complex enzyme are 15-25 ten thousand U/g.
6. The preparation method according to claim 1, wherein the inoculation concentration of the complex probiotics in the step 5) is 107～108CFU/mL, the inoculation amount is 6-8% of the mass of the pumpkin pulp.
7. The preparation method according to claim 1, wherein the fermentation is terminated when the pH of the fermentation liquid in the step 5) is 3.5 to 4.0.
8. The method of any one of claims 1 to 7, comprising the steps of:
1) slicing and blanching: washing fresh and non-rotten pumpkins for 2-3 times by flowing clear water, removing seeds and peels, cutting the pumpkins into uniform slices of 5mm, and blanching the pumpkins in boiling water for 5 min;
2) crushing and pulping: adding purified water with the same volume into the blanched pumpkin slices, adding sodium erythorbate with the total volume of 2 per mill, and pulping to prepare pumpkin pulp;
3) ultrasonic wave and enzyme treatment: adding a complex enzyme preparation into the pumpkin pulp, placing the pumpkin pulp in an ultrasonic crusher for ultrasonic treatment for 10min, and after the ultrasonic treatment is finished, continuing carrying out water bath oscillation enzymolysis for 80min at 55 ℃;
the compound enzyme comprises pectinase, cellulase and papain, the adding mass of each enzyme in the compound enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed pulp respectively, and the ultrasonic intensity is 4W/mL;
the independent enzyme activities of the pectinase, the cellulase and the papain in the compound enzyme are respectively 25 ten thousand U/g, 15 ten thousand U/g and 10 ten thousand U/g;
4) adding a probiotic growth promoter: adding glucose, dipotassium hydrogen phosphate, sodium citrate, manganese sulfate and yeast extract into the enzymatic hydrolysis pumpkin pulp, wherein the adding mass is 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymatic hydrolysis pumpkin pulp respectively;
5) homogenizing: homogenizing the pumpkin pulp added with the probiotic growth promoter under the homogenizing pressure of 20MPa and at the homogenizing temperature of 45 ℃;
6) and (3) sterilization: sterilizing the homogenized pumpkin pulp at 100 deg.C for 10 min;
7) inoculation: sterilizing the pumpkin pulp, quickly cooling to 34 ℃, and mixing the composite probiotic agent according to the proportion of 108Inoculating CFU/mL into pumpkin pulp, wherein the inoculation amount is 8%, fermenting at 34 ℃ for 20h, and finishing fermentation when the fermentation pH value is 4.0.
9. The production method according to any one of claims 1 to 7, comprising the steps of:
1) slicing and blanching: washing fresh non-rotten pumpkins for 2-3 times by flowing clear water, removing seeds and peels, cutting the pumpkins into uniform slices of 5mm, and blanching the pumpkins in boiling water for 5 min;
2) crushing and pulping: adding equal volume of purified water into the blanched pumpkin slices, wherein the total volume of the added purified water is 4 percent0Pulping sodium erythorbate to obtain pulp;
3) ultrasonic wave and enzyme treatment: adding a complex enzyme preparation into the pumpkin pulp, placing the pumpkin pulp in an ultrasonic crusher for ultrasonic treatment for 12min, and after the ultrasonic treatment is finished, continuing carrying out water bath oscillation enzymolysis for 40min at 55 ℃;
the compound enzyme comprises pectinase, cellulase and papain, the adding mass of the compound enzyme is 3 per mill, 5 per mill and 1.5 per mill of the mass of the crushed pulp respectively, and the ultrasonic intensity is 5W/mL;
the independent enzyme activities of the pectinase, the cellulase and the papain in the compound enzyme are respectively 10 ten thousand U/g, 15 ten thousand U/g and 10 ten thousand U/g;
4) adding a probiotic growth promoter: adding glucose, dipotassium hydrogen phosphate, sodium citrate, manganese sulfate and yeast extract into the enzymatic hydrolysis pumpkin pulp, wherein the adding mass is 5%, 0.2%, 0.28%, 0.005% and 0.01% of the mass of the enzymatic hydrolysis pumpkin pulp respectively;
5) homogenizing: homogenizing the pumpkin pulp added with the probiotic growth promoter under the homogenizing pressure of 22MPa and at the homogenizing temperature of 43 ℃;
6) and (3) sterilization: sterilizing the homogenized pumpkin pulp at 103 deg.C for 8 min;
7) inoculation: sterilizing the pumpkin pulp, quickly cooling to 32 ℃, and mixing the composite probiotic agent according to the proportion of 108And (3) inoculating the CFU/mL into the pumpkin pulp, wherein the inoculation amount is 6%, fermenting for 20 hours at 32 ℃, and finishing the fermentation when the fermentation pH value is 3.8.
10. Fermented pumpkin pulp enriched in pumpkin polysaccharides obtainable by the process according to any one of claims 1 to 9.
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