Method for producing enzyme by using apple pomace and molasses and application
Apple pomace and molasses are used as raw materials and are beaten, heated and fermented to prepare apple pomace enzymes, which solves the problem of waste of resources for food processing by-products and achieves efficient production of enzymes and soil improvement effects.
Patent Information
- Application Number
- CN202411702144.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-26
- Publication Date
- 2025-02-25
AI Technical Summary
Food processing by-products such as apple pomace and molasses have not been effectively utilized, resulting in waste of resources. At the same time, existing enzyme production methods rely on fresh fruits and vegetables, which have resource and environmental problems.
Apple pomace is beaten with water and then molasses, and heated under acidic conditions to decompose sucrose into fructose and glucose, providing a carbon source, promoting microbial growth, inoculation of mixed bacteria agents, and fermentation is made of apple pomace enzyme.
The waste recycling of apple pomace and molasses is realized, the active ingredient content of enzymes is improved, the active ingredient of enzymes is enhanced, the soil structure is improved, and the nutrient absorption and yield of plants is increased.
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Figure CN119504321A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of biotechnology, and specifically relates to a method for producing enzymes by utilizing apple pomace and molasses and application thereof. Background Art
[0002] Apple pomace contains rich nutrients such as soluble sugar, vitamins, minerals and cellulose. In addition, apple pomace also contains soluble substances such as pectin, fructose, unsaturated fatty acids and free amino acids. These ingredients make apple pomace a good animal and poultry feed resource. However, except for a small amount of these apple pomace used as fuel, feed and extracting pectin, most of them are treated as garbage, resulting in a huge waste of resources. Molasses is a byproduct of sugar cane sugar production. It is dark brown and viscous. It contains rich sugars (mostly sucrose), polyphenols and other mineral elements, and has antioxidant and anti-inflammatory biological activities. At present, molasses has been widely used in feed production, microbial fermentation, food processing and other industries. After mixing molasses with apple pomace pulp and heating and sterilizing, heating under acidic conditions can decompose sucrose into fructose and glucose, which can supplement the sugar content of apple pomace pulp, provide carbon source for microorganisms, and promote microbial growth and metabolism. Therefore, the production of enzymes using apple pomace and molasses will realize waste recycling and improve the utilization value of apple pomace and molasses.
[0003] Microbial enzymes refer to functional microbial fermentation products that are made from one or more fresh vegetables, fruits, mushrooms, Chinese herbal medicines, etc., fermented by a variety of beneficial microorganisms, and contain rich nutrients such as microorganisms, enzymes, minerals, and secondary metabolites. Bioenzymes can improve soil, establish a good soil microecological environment, and promote crop growth. The microbial metabolites in the enzymes can promote the decomposition of organic matter in the soil and convert it into components that can be directly absorbed and utilized by plants. Therefore, the use of bioenzymes can reduce the use of pesticides and fertilizers and reduce agricultural pollution. At present, plant comprehensive enzymes are widely used in the fields of food and health, and also show unique value in the fields of environmental protection and agriculture. By using the by-products of food processing to produce enzymes, not only can the recycling and high-value utilization of waste be achieved, but fresh fruits and vegetables can also be used to produce enzymes, providing new development ideas for the production of enzyme products, and have broad application prospects. Summary of the invention
[0004] In order to solve the above problems, the present invention provides a method for producing enzyme by utilizing apple pomace and molasses.
[0005] The first object of the present invention is to provide a method for producing enzymes using apple pomace and molasses, comprising the following steps:
[0006] (1) preparing apple pomace pulp: mixing apple peels, cores and pomace and adding water to mix and pulp to obtain apple pomace pulp;
[0007] Preferably, the amount of water added is 2 to 4 times the total mass of the apple peel, core and pomace.
[0008] (2) Add molasses: Add molasses to the apple pomace pulp, stir evenly and heat to sterilize.
[0009] Preferably, the added amount of molasses is 20% to 50% of the total weight of the apple peel, core and pomace.
[0010] Preferably, the sterilization temperature is 80-100°C and the time is 10-20 minutes.
[0011] (3) Inoculation and fermentation: inoculating a mixed bacterial agent and fermenting to obtain an enzyme. The mixed bacterial agent includes plantarum Lactobacillus (CICC21801), fermented mucilaginous Lactobacillus (CICC21835), fermented Pichia pastoris (CICC33372) and rhodotorula glutinosus (CICC31192), and the bacterial agent of the present invention is purchased from the China Industrial Microbiological Culture Collection Management Center.
[0012] Preferably, the mass ratio of each strain in the mixed bacterial agent is 1-2:1-2:1-3:1-3.
[0013] Preferably, the inoculation amount of the mixed bacterial agent is 0.2-0.5 g of mixed bacterial powder / 100 g of fruit pulp.
[0014] Preferably, the fermentation temperature is 25-35° C. and the fermentation time is 3-15 days.
[0015] Preparation of the bacterial agent in step (3):
[0016] 100-200 μL of seed liquid of the plant lactobacillus and fermented mucus lactobacillus is drawn from a freezing tube and inoculated into 100-200 mL of MRS liquid culture medium respectively, activated at 30-37° C. and 150 rpm for 24-48 h, then 1 mL of culture liquid is drawn and inoculated into 500-1000 mL of MRS liquid culture medium for expansion culture, after expansion culture at 30-37° C. and 150 rpm for 48-72 h, bacterial mud is collected by centrifugation, washed twice with sterile water, and then suspended in 10-20 mL of skim milk with a mass fraction of 10-15%, and freeze-dried for 10-20 h at -40° C. and a vacuum degree of 10-30 Pa to obtain freeze-dried bacterial powders of the plant lactobacillus and fermented mucus lactobacillus.
[0017] 100-200 μL of seed solution of the fermented Pichia pastoris and red yeast were drawn from the freezing tube and inoculated into 100-200 mL of YPD liquid culture medium respectively, and activated at 25-30° C. and 200 rpm for 48-72 h, and then 1 mL of culture solution was drawn into 500-1000 mL of YPD liquid culture medium for expansion culture. After expansion culture at 25-30° C. and 200 rpm for 48-96 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10-20 mL of skim milk with a mass fraction of 10-15%, and freeze-dried at -40° C. and a vacuum degree of 10-30 Pa for 10-20 h to obtain freeze-dried bacterial powder of the fermented Pichia pastoris and red yeast.
[0018] The obtained 4 kinds of bacterial powders were transferred into brown ampoules and sealed for storage.
[0019] The second object of the present invention is to provide an apple pomace enzyme prepared by the preparation method of apple pomace enzyme.
[0020] The third object of the present invention is to provide an application of an apple pomace enzyme product in the planting industry.
[0021] Beneficial effects of the present invention:
[0022] The present invention treats apple pomace by adding water to pulp and then adding molasses to inoculate mixed bacterial agents for fermentation, thereby exploring the changes in active chemical components of apple pomace enzyme after fermentation. The raw materials, apple pomace and molasses, are both byproducts of food processing. The addition of molasses can supplement the sugars and active substances in the apple pomace pulp, provide nutrients for microorganisms, and promote the growth of microorganisms. The results showed that the total polysaccharide content in the enzyme increased, and the acidity and sugar content ratio changed significantly after mixed bacterial agent fermentation, indicating that mixed bacterial agent fermentation can significantly increase the active ingredient content of apple pomace enzyme; the plant active ingredients of apple pomace enzyme are increased. In addition, apple pomace enzyme can improve the soil, improve soil compaction, and promote plant absorption of nutrients. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 The active ingredient changes of apple pomace enzyme in different implementation modes are shown in FIG.
[0024] Figure 2 The quality change diagram of the dishes in different treatment groups in the embodiment DETAILED DESCRIPTION
[0025] The present invention is further described in detail below in conjunction with specific embodiments and accompanying drawings, but it should be understood that the examples given are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way. Unless otherwise specified, the experimental methods used in the following examples are all conventional methods, and the materials, reagents, etc. used can be obtained through commercial channels.
[0026] Example 1
[0027] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0028] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0029] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0030] (3) Preparation of bacterial agents: 100 μL of seed solution of Lactobacillus plantarum and Lactobacillus fermentans were drawn from cryopreservation tubes and inoculated into 100 mL of MRS liquid culture medium respectively, and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powders of Lactobacillus plantarum and Lactobacillus fermentans were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0031] 100 μL of seed solution from the above-mentioned fermented Pichia pastoris and red yeast was drawn from the freezing tube and inoculated into 100 mL YPD liquid culture medium respectively, and activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture solution was drawn into 500 mL YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The fermented Pichia pastoris and red yeast freeze-dried bacterial powder were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0032] (4) Inoculation and fermentation: The four kinds of bacterial powder were evenly mixed in a mass ratio of 1:1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.5 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0033] Example 2
[0034] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0035] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 400 mL of water for pulping to obtain apple pomace pulp;
[0036] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0037] (3) Preparation of bacterial agents: 100 μL of seed solution of Lactobacillus plantarum and Lactobacillus fermentans were drawn from cryopreservation tubes and inoculated into 100 mL of MRS liquid culture medium respectively, and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powders of Lactobacillus plantarum and Lactobacillus fermentans were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0038] 100 μL of seed solution from the above-mentioned fermented Pichia pastoris and red yeast was drawn from the freezing tube and inoculated into 100 mL YPD liquid culture medium respectively, and activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture solution was drawn into 500 mL YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The fermented Pichia pastoris and red yeast freeze-dried bacterial powder were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0039] (4) Inoculation and fermentation: The four kinds of bacterial powder were evenly mixed in a mass ratio of 1:1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.5 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0040] Example 3
[0041] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0042] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0043] (2) Add molasses: Add 40 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0044] (3) Preparation of bacterial agents: 100 μL of seed solution of Lactobacillus plantarum and Lactobacillus fermentans were drawn from cryopreservation tubes and inoculated into 100 mL of MRS liquid culture medium respectively, and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powders of Lactobacillus plantarum and Lactobacillus fermentans were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0045] 100 μL of seed solution from the above-mentioned fermented Pichia pastoris and red yeast was drawn from the freezing tube and inoculated into 100 mL YPD liquid culture medium respectively, and activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture solution was drawn into 500 mL YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The fermented Pichia pastoris and red yeast freeze-dried bacterial powder were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0046] (4) Inoculation and fermentation: The four kinds of bacterial powder were evenly mixed in a mass ratio of 1:1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.5 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0047] Example 4
[0048] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0049] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0050] (2) Add molasses: Add 100g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 minutes:
[0051] (3) Preparation of bacterial agents: 100 μL of seed solution of Lactobacillus plantarum and Lactobacillus fermentans were drawn from cryopreservation tubes and inoculated into 100 mL of MRS liquid culture medium respectively, and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powders of Lactobacillus plantarum and Lactobacillus fermentans were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0052] 100 μL of seed solution from the above-mentioned fermented Pichia pastoris and red yeast was drawn from the freezing tube and inoculated into 100 mL YPD liquid culture medium respectively, and activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture solution was drawn into 500 mL YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The fermented Pichia pastoris and red yeast freeze-dried bacterial powder were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0053] (4) Inoculation and fermentation: The four kinds of bacterial powder were evenly mixed in a mass ratio of 1:1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.5 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0054] Example 5
[0055] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0056] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0057] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0058] (3) Preparation of bacterial agents: 100 μL of seed solution of Lactobacillus plantarum and Lactobacillus fermentans were drawn from cryopreservation tubes and inoculated into 100 mL of MRS liquid culture medium respectively, and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powders of Lactobacillus plantarum and Lactobacillus fermentans were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0059] 100 μL of seed solution from the above-mentioned fermented Pichia pastoris and red yeast was drawn from the freezing tube and inoculated into 100 mL YPD liquid culture medium respectively, and activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture solution was drawn into 500 mL YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The fermented Pichia pastoris and red yeast freeze-dried bacterial powder were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0060] (4) Inoculation and fermentation: The four kinds of bacterial powder were evenly mixed in a mass ratio of 1:1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.2 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0061] Comparative Example 1
[0062] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0063] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0064] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0065] (3) Preparation of bacterial agent: Lactobacillus plantarum (CICC21801) was inoculated into 100 mL of MRS liquid culture medium and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was aspirated and inoculated into 500 mL of MRS liquid culture medium for expansion. After expansion for 48 h under the same conditions, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk. The lyophilized powder of Lactobacillus plantarum was obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0066] (4) Inoculation and fermentation: Inoculate Lactobacillus plantarum at an inoculation rate of 0.5 g bacterial powder / 100 g fruit pulp and ferment at 30°C for 10 days.
[0067] Comparative Example 2
[0068] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0069] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0070] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0071] (3) 100 μL of seed solution of Rhodotorula glutinosus (CICC31192) was aspirated from the cryopreservation tube and inoculated into 100 mL of YPD liquid medium, and activated at 30°C and 200 rpm for 48 h. Then 1 mL of culture solution was aspirated and inoculated into 500 mL of YPD liquid medium for expansion. After 48 h of expansion under the same conditions, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk. The freeze-dried powder of Rhodotorula glutinosus was obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0072] (4) Inoculation and fermentation: Inoculate red yeast at a rate of 0.5 g of bacterial powder per 100 g of fruit pulp and ferment at 30°C for 10 days.
[0073] Comparative Example 3
[0074] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0075] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0076] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0077] (3) Preparation of bacterial agents: 100 μL of seed solution of Lactobacillus plantarum and Lactobacillus fermentans were drawn from cryopreservation tubes and inoculated into 100 mL of MRS liquid culture medium respectively, and activated at 37°C, 150 rpm for 24 h. Then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powders of Lactobacillus plantarum and Lactobacillus fermentans were obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0078] 100 μL of seed liquid was drawn from the cryopreservation tube and inoculated into 100 mL of YPD liquid culture medium, activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture liquid was drawn into 500 mL of YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powder of red yeast was obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0079] (4) Inoculation and fermentation: The three kinds of bacterial powder were mixed evenly in a weight ratio of 1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.5 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0080] Comparative Example 4
[0081] A preparation method for producing enzymes by using apple pomace and molasses, comprising:
[0082] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0083] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp, stir evenly, and sterilize at 85°C for 15 min:
[0084] (3) Preparation of bacterial agent: 100 μL of seed solution of fermented mucus lactobacillus was drawn from the cryopreservation tube and inoculated into 100 mL of MRS liquid culture medium, activated at 37°C, 150 rpm for 24 h, and then 1 mL of culture solution was drawn into 500 mL of MRS liquid culture medium for expansion culture. After expansion culture at 37°C, 150 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The fermented mucus lactobacillus freeze-dried powder was obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0085] 100 μL of seed solution of fermented Pichia pastoris and Rhodotorula glutinosus was drawn from the freezing tube and inoculated into 100 mL of YPD liquid culture medium respectively, and activated at 30°C and 200 rpm for 48 h, and then 1 mL of culture solution was drawn into 500 mL of YPD liquid culture medium for expansion culture. After expansion culture at 30°C and 200 rpm for 48 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10 mL of 10% skim milk by mass fraction. The freeze-dried bacterial powder of fermented Pichia pastoris and Rhodotorula glutinosus was obtained at -40°C and a vacuum degree of 10 Pa for 10 h.
[0086] (4) Inoculation and fermentation: The three kinds of bacterial powder were mixed evenly in a mass ratio of 1:1:1, and the mixed bacterial agent was inoculated at an inoculation rate of 0.5 g of mixed bacterial powder / 100 g of fruit pulp, and fermented at 30°C for 10 days.
[0087] Comparative Example 5
[0088] A preparation method for producing enzyme by utilizing apple pomace and molasses, comprising:
[0089] (1) preparing apple pomace pulp: mixing 200 g of peel, core and pomace and adding 800 mL of water for pulping to obtain apple pomace pulp;
[0090] (2) Add molasses: Add 70 g of molasses to the apple pomace pulp and mix well. Ferment naturally at 30°C for 10 days.
[0091] Example 8
[0092] Soil Improvement Testing
[0093] The apple pomace enzyme in Example 1 of the present invention was selected for soil improvement and planting experiments, 40 kg of urea per mu was added twice, and 30 cubic meters of water was applied per mu each time. 0.5 square meters was selected for each group of the experiment, and a total of 3 vegetables were used in each group for the experiment.
[0094] Table 1: Experimental groups
[0095]
[0096] The apple pomace enzymes prepared in Examples 1 to 5 and Comparative Examples 1 to 5 were tested, and the test results are shown in Table 2 below:
[0097] sample Total acid (g / L) Total sugar (g / L) Protein content (g / L) Example 1 4.25 15.66 17.33 Example 2 3.65 20.54 10.21 Example 3 3.05 5.28 10.06 Example 4 2.95 22.87 12.42 Example 5 3.20 18.69 10.75 Comparative Example 1 3.79 18.66 8.25 Comparative Example 2 2.47 15.02 9.32 Comparative Example 3 3.61 14.83 9.11 Comparative Example 4 3.35 13.92 9.75 Comparative Example 5 2.08 30.67 6.30
[0098] From the test results, it can be seen that the total protein content in the apple pomace enzyme obtained in Examples 1 to 5 of the present invention is higher than that in the apple pomace enzyme of Comparative Examples 1 to 5, indicating that the enzyme and nitrogen-containing active component content obtained by the apple pomace molasses enzyme fermented by four kinds of microorganisms are higher. However, in Examples 1 to 5, the ratio of apple pomace to water, the amount of molasses added, and the amount of bacterial agent inoculation all have a significant effect on the fermentation results. Therefore, Example 1 is the group with the best indicators, that is, the active component content of apple pomace enzyme can be increased by mixed bacterial agent fermentation.
[0099] Test Example 8, after growing for 65 days, the crops were harvested and weighed, and the data are shown in Table 3 below:
[0100]
[0101] As shown in the above table, the apple pomace enzyme can improve the soil, reduce soil compaction, promote plant absorption of nutrients, and increase plant yields by 22.10%.
[0102] The above description has fully disclosed the specific embodiments of the present invention. It should be pointed out that any changes made by those skilled in the art to the specific embodiments of the present invention do not deviate from the scope of the claims of the present invention. Accordingly, the scope of the claims of the present invention is not limited to the above specific embodiments.
Claims
1. A method for producing enzymes using apple pomace and molasses, characterized in that: The steps include: (1) preparing apple pomace pulp: mixing apple peels, cores and pomace and adding water to mix and pulp to obtain apple pomace pulp; (2) Adding molasses: adding molasses to the apple pomace pulp, stirring evenly and heating to sterilize; (3) Inoculation and fermentation: inoculating a mixed bacterial agent and fermenting to obtain an enzyme; the mixed bacterial agent includes plant lactobacillus CICC21801, fermented lactobacillus CICC21835, fermented Pichia pastoris CICC33372 and Rhodotorula glutinosus CICC31192.
2. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: In step (1), the amount of water added is 2 to 4 times the total mass of the apple peel, core and pomace.
3. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: In step (2), the amount of molasses added is 20% to 50% of the total weight of the apple peel, core and pomace.
4. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: In step (2), the sterilization temperature is 80-100°C and the time is 10-20 minutes.
5. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: In step (3), the mass ratio of each strain in the mixed bacterial agent is 1-2:1-2:1-3:1-3.
6. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: In step (3), the inoculation amount of the mixed bacterial agent is 0.2-0.5 g of mixed bacterial powder / 100 g of fruit pulp.
7. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: In step (3), the fermentation temperature is 25 to 35° C. and the fermentation time is 3 to 15 days.
8. The method for producing enzymes using apple pomace and molasses according to claim 1, characterized in that: Preparation of the bacterial agent in step (3): 100-200 μL of seed solution of the plant lactobacillus and fermented mucus lactobacillus are drawn from a freezing tube and inoculated into 100-200 mL of MRS liquid culture medium respectively, activated at 30-37° C. and 150 rpm for 24-48 h, then 1 mL of culture solution is drawn and inoculated into 500-1000 mL of MRS liquid culture medium for expansion culture, and after expansion culture at 30-37° C. and 150 rpm for 48-72 h, bacterial mud is collected by centrifugation, washed twice with sterile water, and then suspended in 10-20 mL of skim milk with a mass fraction of 10-15%, and freeze-dried at -40° C. and a vacuum degree of 10-30 Pa for 10-20 h to obtain freeze-dried bacterial powders of plant lactobacillus and fermented mucus lactobacillus; 100-200 μL of the seed solution of the fermented Pichia pastoris and red yeast were drawn from the cryopreservation tube and inoculated into 100-200 mL of YPD liquid culture medium respectively, activated at 25-30° C. and 200 rpm for 48-72 h, then 1 mL of the culture solution was drawn and inoculated into 500-1000 mL of YPD liquid culture medium for expansion culture, and after expansion culture at 25-30° C. and 200 rpm for 48-96 h, the bacterial sludge was collected by centrifugation, washed twice with sterile water, and then suspended in 10-20 mL of skim milk with a mass fraction of 10-15%, and freeze-dried at -40° C. and a vacuum degree of 10-30 Pa for 10-20 h to obtain freeze-dried bacterial powder of the fermented Pichia pastoris and red yeast; The obtained 4 kinds of bacterial powders were transferred into brown ampoules and sealed for storage.
9. An apple pomace enzyme obtained by the method for producing enzyme using apple pomace and molasses as described in any one of claims 1 to 8.
10. The use of apple pomace enzyme obtained by the method for producing enzyme using apple pomace and molasses according to any one of claims 1 to 8, characterized in that: Application and planting industry.