Preparation method and application of fruit and vegetable environment-friendly enzyme

Abstract

The invention discloses a preparation method and application of a fruit and vegetable environment-friendly enzyme, application of the fruit and vegetable environment-friendly enzyme in improving growth performance of fruit trees and crops, and particularly application of the fruit and vegetable environment-friendly enzyme in replacing chemical fertilizers and pesticides. The production method of the fruit and vegetable environment-friendly enzyme comprises the following steps: (1) selecting materials; (2) environment disinfection; (3) equipment disinfection; (4) culture medium formulation; (5) production feeding; (6) directional fermentation; (7) tank releasing: after the fermentation time expires, clarification of fermentation liquid, decomposition and sedimentation of fruits and vegetables, tank releasing, opening of a sampling port valve of the fermentation tank during fermentation tank releasing, and sampling for inspection; (8) detecting items; and (9) performing solid-liquid separation to obtain an enzyme product. The enzyme product obtained through natural fermentation of fruits and vegetables has a very good effect in the aspect of growth performance of fruit trees and crops, so that the enzyme product can replace chemical fertilizers and pesticides, is beneficial to improving the eating taste of the fruits and the crops, and can also reduce pesticide residues of the fruits and the crops.

Description

Technical Field

[0001] The invention relates to the technical field of enzyme production, and in particular to a method for producing an environmentally friendly fruit and vegetable enzyme and application thereof. Background Art

[0002] Environmentally friendly enzymes are products containing specific biologically active ingredients that are made from plant raw materials, with or without auxiliary materials, and through microbial fermentation. They can be used in multiple environmental protection industries such as odor control, water purification, and soil improvement, and have been widely promoted and applied at home and abroad. The use of fruit by-products to produce environmentally friendly enzymes provides a new way for their resource utilization. In recent years, some scholars have successfully used fruit by-products to prepare environmentally friendly enzymes. For example: Wang Yang et al. used defective fruits and discarded vegetables to ferment naturally for 12 months to prepare environmentally friendly enzymes and applied them to aquaculture. They have strong in vitro inhibitory and lethal effects on Gram-negative pathogenic microorganisms; Bu Junzhi et al. used orange peels, apple peels, cucumbers and other raw materials to prepare environmentally friendly enzymes after 3 months of fermentation. After testing, they showed that they had protease, lipase and amylase activities and could be used for household cleaning.

[0003] The performance of each batch of enzyme products obtained by the existing production process of environmentally friendly enzymes is unstable, and they have not been applied in the fields related to fruit tree or crop planting.

[0004] Based on this, the present invention designs a method for preparing an environmentally friendly fruit and vegetable enzyme and its application to solve the above problems. Summary of the invention

[0005] The purpose of the present invention is to provide a method for preparing an environmentally friendly fruit and vegetable enzyme and its application to solve the problems raised in the above background technology.

[0006] To achieve the above object, the present invention provides the following technical solution: a method for preparing environmentally friendly fruit and vegetable enzymes, the specific steps are as follows:

[0007] (1) Material selection: Select fruits and vegetables of the current season, clean them and dry them;

[0008] (2) Environmental disinfection: spray the environment with a solution of Chlorhexidine and use ultraviolet light to disinfect for 15-60 minutes;

[0009] (3) Equipment disinfection: Disinfect the fermentation tank with 70-75% alcohol as a disinfectant;

[0010] (4) Culture medium formula: 7%-15% brown sugar, 20%-40% fruits and vegetables, 50%-73% water, pH is neutral, and the culture medium is prepared;

[0011] (5) Production and feeding: Crush the fruits and vegetables and put them into the fermentation tank, the final volume of which shall not exceed 75%-80% of the space of the fermentation tank, package them, and label them. The label shall have the production date;

[0012] (6) Directed fermentation: Use the dominant bacteria of fruits and vegetables to transform microorganisms and ferment naturally in a cool place for 2-4 months;

[0013] (7) Release the tank: After the fermentation time expires, the fermentation liquid is clear and the fruits and vegetables are decomposed and settled, and the tank can be released. When releasing the fermentation tank, open the sampling port valve of the fermentation tank and take samples for inspection;

[0014] (8) Test items: Take enzyme samples to test their properties and sensory indicators, general physical and chemical indicators of pH and TDS, and characteristic physical and chemical indicators of total acid, organic acid, and total bacterial count;

[0015] (9) Solid-liquid separation: The fermentation liquid that meets the inspection standards is filtered through a 40-80 mesh screen or centrifuged in a box centrifuge to obtain the liquid enzyme product.

[0016] Preferably, the fruits and vegetables in step (1) are seasonal fruit and vegetable waste collected from the market, with rotten and spoiled fruits and vegetables removed.

[0017] Preferably, in step (6), the exhaust is performed once every 3-4 days within half a month after the start of fermentation.

[0018] An application of an environmentally friendly fruit and vegetable enzyme, and the application of the environmentally friendly fruit and vegetable enzyme in improving the growth performance of fruit trees and crops, specifically the application of the environmentally friendly fruit and vegetable enzyme to replace chemical fertilizers and pesticides.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention can improve the performance uniformity of each batch of the environmentally friendly enzyme product by disinfecting the environment and equipment and placing the cans for inspection, so that the product is conducive to large-scale production and market promotion; the enzyme product obtained by natural fermentation of fruits and vegetables in the present invention has a very good effect in the application of the growth performance of fruit trees and crops, so the enzyme product can replace chemical fertilizers and pesticides, is beneficial to improving the edible taste of fruits and crops, and can also reduce pesticide residues in fruits and crops.

[0020] Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for describing the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0022] Figure 1 Comparison of loquat tree diseases of the present invention;

[0023] Figure 2 The inhibitory activity of naturally fermented loquat enzyme against Candida albicans;

[0024] Figure 3 The antibacterial activity of naturally fermented loquat enzyme against Malassezia furfur;

[0025] Figure 4 The antibacterial activity of naturally fermented loquat enzyme against Bacillus aeruginosa

[0026] Figure 5 The antibacterial activity of naturally fermented loquat enzyme against Staphylococcus aureus. DETAILED DESCRIPTION

[0027] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0028] 1: Research on the production process of environmentally friendly enzymes

[0029] Main reagents and raw materials

[0030] Loquat: Xianyou Qingfeng Ecological Farm Co., Ltd. Brown sugar (food grade): Guangxi Mashan Nanhua Sugar Co., Ltd.; YPD medium and MRS medium: Guangdong Huankai Microbiological Technology Co., Ltd.

[0031] Main instruments and equipment

[0032] LC-2030 high performance liquid chromatograph: Shimadzu Corporation, Japan; Allegra X-15R centrifuge: Beckman Company, Germany; 752 UV spectrophotometer: Shanghai Precision Scientific Instrument Co., Ltd.; DKZ-2B electric constant temperature water bath: Shanghai Yiheng Technology Co., Ltd.; S20K acidity meter: Mettler-Toledo Company.

[0033] Test methods

[0034] Preparation of loquat enzyme: Wash loquats 2-3 times with tap water and purified water, dry them in a clean place, slice them, remove the core, prepare them according to different ratios of sugar: loquat: water, and ferment them. Take samples at intervals, and store the supernatant after centrifugation in a -80℃ ultra-low temperature refrigerator for testing.

[0035] Index determination

[0036] Determination of effective viable bacterial count: Determine according to the method specified in NY / T 2321. Take 5 mL of the sample and add it to 100 mL of sterile water with glass beads. Let it stand for 20 minutes and fully shake it on a rotary shaker at 200 r / min for 30 minutes to form a mother liquid bacterial suspension. Use a 1 mL sterile transfer tube to draw 0.5 mL of the above mother liquid bacterial suspension and add it to 4.5 mL of sterile water. Perform a series of dilutions at a ratio of 1:10 to obtain 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 Dilution multiples of bacterial suspension. Take 3 consecutive appropriate dilutions for each sample, use a 0.2mL sterile pipette to take 0.1mL of bacterial suspension of different dilutions, add them to the pre-prepared solid culture medium plate, and evenly spread the bacterial suspension of different dilutions on the surface of the plate. Repeat each dilution 3 times, and use sterile water as a blank control, and culture under appropriate conditions.

[0037]

[0038] pH value and total acid determination: pH value is determined according to the method specified in GB / T 10468. Preheat the pH meter, calibrate the instrument with standard solution, and then determine the pH of the sample suspension. Record the reading after the instrument is stable. The determination method of total acid content refers to GB12456-2021 "Determination of Total Acid in Food". Pipette 0.5mL of test solution into a 150mL conical flask with a condenser, add about 25mL of carbon dioxide-free water, mix well, place in a boiling water bath and boil for 30min, cool to room temperature and dilute to 50mL with carbon dioxide-free water for determination. Use a pipette to draw 25mL of test solution, cool to room temperature, add it to a 250mL conical flask, add 2 drops of phenolphthalein indicator solution, and titrate with 0.01mol / L sodium hydroxide titration solution until it is slightly red and does not fade for 30s. Record the volume value of the sodium hydroxide titration solution. According to the above operation, use the same volume of carbon dioxide-free water instead of the test solution for a blank test, and record the volume value of the consumed sodium hydroxide titration solution. The total acid content in the sample is calculated according to the following formula:

[0039] X=[c×(V1-V2)×k×F÷m]×1000

[0040] X: total acid content in the sample (g / L); c: concentration of sodium hydroxide standard titration solution (mol / L); V1: volume of sodium hydroxide standard titration solution consumed when titrating the test solution (mL); V2: volume of sodium hydroxide standard titration solution consumed in the blank test (mL); k: acid conversion factor (lactic acid: 0.09); F: dilution factor of the sample; m: mass of the sample (mL); 1000: conversion factor.

[0041] Determination of polysaccharide content: Determine according to the method specified in SN / T 4260. Pipette 0, 0.2, 0.4, 0.6, 0.8, 1mL of 100mg / mL glucose solution into 20mL stoppered test tubes, and fill to 1mL with distilled water. Add 1mL of 5% phenol solution to each test tube, and then quickly add 5mL of concentrated sulfuric acid. After standing for 10min, place the test tube in a 30℃ water bath to react for 20min. Take a sample and measure the optical density at a wavelength of 490nm by spectrophotometry (use pure water instead of standard solution as a blank control). The horizontal axis is the glucose concentration, and the vertical axis is the optical density value. Draw a standard curve. Sample detection: Use a pipette to transfer 1mL of sample solution to a stoppered test tube, and measure the optical density of the solution in the same way, and calculate its polysaccharide content according to the standard curve.

[0042] Determination of organic acid (lactic acid): Determine according to the method specified in GB 5009.157. Weigh 2.5g of lactic acid into a 50mL volumetric flask, dilute to volume, and prepare a 5000μg / mL lactic acid standard stock solution. Pipette 0.5, 1, 2, 5, and 10mL of the lactic acid standard stock solution into a 25mL volumetric flask, dilute to the scale with 0.1% phosphoric acid solution, prepare the lactic acid standard curve working solution, and store at 4°C. Start sampling and analysis after the HPLC is stable.

[0043] The chromatographic column and chromatographic operating conditions are as follows: the chromatographic column is Kromasil100-5-C18 (4.6mm×250mm), the mobile phase is a mobile phase with a ratio of 0.1% phosphoric acid solution to methanol (97.5%-2.5%), the flow rate is 0.6mL / min, the column temperature is 40°C, the injection volume is 20μL, and the detection wavelength is 210nm; a standard curve is drawn with the chromatographic peak area as the ordinate and the concentration of the standard working solution as the abscissa.

[0044] Sample detection: After the fermentation sample was diluted 10 times with deionized water, the test solution filtered through a 0.22 μm microporous filter membrane was used to determine its organic acid content by HPLC to obtain the peak area, and the concentration of organic acid in the test solution was obtained according to the standard curve.

[0045] Investigation on the amount of brown sugar added

[0046] Preliminary investigations found that brown sugar is more suitable for fermentation, and the effects of different proportions of brown sugar on environmentally friendly enzyme fermentation were investigated. The test plan was designed according to different proportions of brown sugar-pulp-water. When the initial pulp content was 30%, brown sugar addition was tested at 5%, 10% and 15% to produce three groups of enzymes. During the fermentation process, factors such as pH, polysaccharide content, organic acid content and effective viable bacteria were continuously monitored as indicators to investigate the effects of different sugar additions on fermentation.

[0047] In a 1L fermenter, the pH changes during the enzyme fermentation process with three different ratios of ingredients were recorded. The results are shown in Table 1. The pH of the loquat enzyme fermentation with different carbon source test groups was relatively stable during the fermentation process, and the pH value of the fermentation liquid was stable between pH 3 and 4 until the end of fermentation.

[0048] Table 1 pH test results of fermentation process with different brown sugar ratios in 1L fermenter

[0049]

[0050] In a 1L fermentation bottle, the changes in the number of effective live bacteria during the fermentation process of the three brown sugar content enzymes were recorded. The results are shown in Table 2. The changes in the number of effective live bacteria during the fermentation process of loquat enzyme fermentation with different brown sugar content test groups were different. The number of effective live bacteria in the 5% brown sugar group in the early and middle stages of fermentation was 10 7 Order of magnitude, maintained at 10 in the middle and late stages of fermentation 6 The number of effective live bacteria in the 10% brown sugar group was relatively stable, and the number of effective live bacteria remained at 10 during the entire fermentation cycle. 8 In terms of order of magnitude, the effective viable bacteria count of the 15% brown sugar group was relatively low, and the effective viable bacteria count remained at 10 throughout the fermentation cycle. 4 ~10 5 This indicates that 10% brown sugar is more suitable and conducive to the growth of loquat enzyme flora and maintains a stable microbial ecological environment; high concentrations of brown sugar are not conducive to the growth of flora.

[0051] Table 2 Results of effective viable bacteria count during fermentation in 1L fermenter with different amounts of brown sugar added

[0052]

[0053] In a 1L fermenter, the changes in crude polysaccharide content during enzyme fermentation in three test groups with different brown sugar contents were recorded. The results are shown in Table 3. The initial sugar contents were 5%, 10% and 15% respectively. With the progress of fermentation, the sugar consumption rates in each group were different: the 5% brown sugar test group consumed faster during 0-15 days, and the sugar content decreased to 2.8%, and the concentration change gradually decreased during 25-145 days, indicating that there was no longer a large amount of sugar consumption during this period, the microbial growth tended to be stable, and the fermentation was basically over; the 10% brown sugar test group consumed less than the 5% brown sugar group during 0-15 days, and the sugar content decreased to 9.1%, and the sugar content gradually decreased during 25-145 days, indicating that a large amount of sugar was consumed during this period, the microbial growth was in good condition, and the polysaccharide content did not change significantly in the late fermentation period (115-145 days), indicating that the microbial growth tended to be stable and the fermentation was basically over; the 15% brown sugar test group consumed slower during 0-145 days, and the sugar content gradually decreased to 13.6% during the whole fermentation process, and the polysaccharide content changed very little. It is speculated that the high concentration of sugar content is not suitable for microbial growth.

[0054] Table 3 Polysaccharide detection results during the fermentation process of 1L fermenter with different amounts of brown sugar added

[0055]

[0056]

[0057] In a 1L fermentation tank, the changes in organic acid content during the fermentation process of the three test groups with different brown sugar contents were recorded, and the results are shown in Table 4. The results show that the changes in organic acid content in the three test groups with different brown sugar contents showed different trends. Among them, the organic acid content of the 5% brown sugar test group was relatively low, ranging from 2.02 to 4.47 g / L. In the early and middle stages of fermentation (15 to 55 days), the organic acid content was between 3.31 and 4.47 g / L, and the organic acid content dropped to 2.02 g / L at 55 to 140 days of fermentation; the organic acid content of the 10% brown sugar test group showed a trend of gradual decrease, and the organic acid content gradually increased from 15 to 25 days of fermentation. The organic acid content in the 15% brown sugar test group was relatively high, reaching 12.89 g / L in the early fermentation period (15-25 days), and decreased in the middle and late fermentation period (25-145 days), ranging from 8.16 to 12.89 g / L.

[0058] Table 4 Results of organic acid detection during fermentation in 1L fermenter with different amounts of brown sugar added

[0059]

[0060] Taking factors such as pH, crude polysaccharide content, organic acid content and effective live bacteria count in the fermentation liquid as indicators, the investigation results showed that 10% brown sugar was more conducive to enzyme fermentation.

[0061] 2: Environmentally friendly enzymes improve fruit tree growth performance and fruit application

[0062] Taking loquat trees as the research object, trees of the same variety with different numbers were selected as the control group and experimental group for experiments. The control group was cultivated conventionally, and the experimental group was sprayed with enzymes. The effects of environmentally friendly enzymes on loquat yield and quality were investigated, and the differences in production performance indicators and fruit quality-related indicators between the control group and the experimental group were analyzed.

[0063] method

[0064] Loquat planting experiment: experimental group and control group were set up in different areas, and two areas far away from the pond and separated by hills were set as experimental area and control area respectively. The routine operations of experimental area and control area were the same, including pruning, flower combing, fruit combing and mowing twice a year. In the control area, 5kg of compound fertilizer and urea and other fertilizers were applied per tree in May each year, and herbicides were used for weeding once. In the experimental area, environmentally friendly enzymes made from waste fruits such as defective loquats were used to replace compound fertilizers, urea and herbicides. In principle, they were sprayed once a month, and about 0.25kg of enzyme filtrate was sprayed on each tree each time (diluted to 20 times with water and sprayed on the fruit trees). Spraying was stopped when flowering, harvesting, rainy days, etc.

[0065] Results and Analysis

[0066] The comparison of loquat fruit yield for three consecutive years in 2022, 2023 and 2024 is shown in Table 5. Without the use of compound fertilizers and urea, the results show that under the same planting environment, the loquat yield of the experimental group is equivalent to that of the control group. The specific situation is: in 2022, the yield of the experimental group is equivalent to that of the control group, the yield of the experimental group in 2023 increased by 6.5%, and the yield of the experimental group in 2024 increased by 8.3%; the insect pest situation of the fruit trees in the two groups is similar, and the specific situation cannot be calculated by equal scoring per tree. The insect pest distribution is uneven, but the results of the two groups are not much different, and there is no obvious difference in the number and type of pests caught by the insect trap; the disease situation is also unevenly distributed and cannot be displayed with specific data, but compared with the overall planting area, the control area has more leaf spot diseases, thinner leaves, and some leaves are curled and wrinkled. The loquat leaves in the enzyme group are relatively large, the leaves are flat and unfolded, and there are fewer spotted leaves. The disease situation is shown in Table 5. Figure 1. The results of the pesticide residue detection of loquat fruits in 2022 and 2023 showed that no relevant pesticide residues such as methyl parathion, chlorpyrifos, hexachlorvos, dimethoate, and acephate were detected. The quality inspection reports of loquat fruits in 2023 and 2024 showed that the total phenol, total sugar, and vitamin C contents of the loquat fruits in the enzyme test group were higher than those in the control group, as shown in Table 6; the appearance test area was more complete, substantial, and fuller than the control area, and the taste was also better, and it received praise after being tasted by experts.

[0067] Yield is an important indicator to measure the production capacity of loquat trees, and fertilization is an important way to ensure loquat yield. Conventional farming requires at least 10kg of compound fertilizer per loquat tree per year. Studies have shown that the application of plant enzymes in agricultural planting has a good yield-increasing effect, which can increase crop fruiting rate and yield, and can also improve fruit quality and increase the content of key quality indicators such as soluble sugar and vitamins in fruits. The organic acids, polyphenols, and amino acids contained in the inferior loquat environmental enzymes used in this experiment are all beneficial ingredients for crop growth and can effectively promote crop growth. This may be the reason why the fruit trees in the experimental group still maintain a considerable yield after the environmental enzymes replace compound organic fertilizers, and even gradually increase the yield of loquats. By comparison, it was found that the fruit quality of the enzyme group was higher than that of the control group. By using enzyme fertilizers instead of compound fertilizers, weight loss and efficiency can be achieved while improving quality.

[0068] Common spot diseases of loquat trees are caused by fungal pathogens. Leaf spot diseases mainly overwinter in the form of conidia and mycelium on the damaged trees of virus leaves or virus fruits. They are prone to occur in warm and humid environments, and invade and infect many times every year. The frequent use of pesticides has increased the resistance of plant crops to pests and diseases, polluting the environment and reducing the effectiveness of pest control. Environmentally friendly enzymes are weakly acidic, which is not conducive to the growth of pathogens. At the same time, the enzyme fermentation liquid contains rich proteases, total phenols and other active substances that have a certain inhibitory effect on pathogens. For example, β-glucanase can specifically act on the cell wall of plant pathogenic fungi, thereby inhibiting the growth and reproduction of fungi. It is an important plant antifungal substance.

[0069] Table 5 Comparison of loquat fruit production from 2022 to 2024

[0070]

[0071] Table 6 Data table of loquat fruit related quality indicators

[0072]

[0073] Note: The total phenol content was calculated by drawing a standard curve with gallic acid (GA), and the result was expressed as g.GA / kg.

[0074] summary

[0075] Taking loquat trees as the research object, loquat trees of the same variety but different numbers were selected as the control group and experimental group for the experiment. The control group was cultivated by conventional farming methods, and the experimental group was sprayed with enzymes. The effects of different process enzymes on loquat yield and quality were investigated, and the differences in production performance indicators and fruit quality-related indicators between the control group and the experimental group were analyzed. The results showed that the loquat yield of the experimental group was higher than that of the control group; the results of pesticide residue detection for two consecutive years met the requirements, the loquat fruit appearance experiment showed that the experimental group was more complete, substantial, and fuller than the control group, and the taste was better, and the experts gave good evaluations; the loquat fruit detection report showed that the total phenols, total sugars, VC and other detection indicators of the experimental group were higher than those of the control group; the loquat tree experimental group had fewer diseases, the loquat leaves were relatively large, and there were fewer spotted leaves.

[0076] 3. Application of environmentally friendly enzymes to improve crop growth performance

[0077] Rice was used as the research object and divided into a control group and an experimental group. The experimental group was only sprayed with enzymes and the amount of chemical fertilizers was reduced, while the control group used fertilizers and pesticides according to conventional methods to assess rice yield and disease conditions.

[0078] method:

[0079] Rice planting experiment: Experimental and control groups were set up in different areas, separated by village roads. The breeding and transplanting steps were the same, and they were uniformly managed by Nanxing Village, the test site. The water and fertilizer management time was consistent. The experimental group sprayed enzymes for the first time one month after transplanting, and then sprayed once every month. The enzyme stock solution was 10 kg per mu, diluted 20-50 times, and sprayed with a high-pressure car washer. Four times in one planting cycle. The control group applied 25 catties of urea and 25 catties of compound fertilizer 7-8 days after transplanting, and the experimental group did not apply chemical fertilizers. The conventional group applied 20 catties of potassium fertilizer and 30 catties of compound fertilizer at 15-20 days, and the experimental group was consistent. The control group used thiazide isoprocarb and chlorpyrifos at the same time at 15-20 days, and the experimental group only sprayed enzymes without pesticides. The control group found pests about two months after planting, and used chlorpyrifos and carbendazim for prevention and control, and the experimental group sprayed enzymes. One month before harvest, that is, about three months after planting, the control group had serious diseases, so Jinggang tricyclazole and benzoprocyclazole were used again. The disease in the experimental group was mild, so the enzyme was sprayed again.

[0080] Each experimental plot was sampled at five points, with 10 rice plants surveyed at each point and the fourth leaf of each rice plant surveyed.

[0081] The survey method is based on the "Guidelines for Field Efficacy Tests of Pesticides" and the rice blast leaf blast classification survey standards:

[0082] 0: No disease

[0083] Level 1: Brown spots as small as a pinpoint

[0084] Grade 3: small, round or slightly elongated brown necrotic gray spots, 1-2 mm in diameter

[0085] Level 5: Typical rice blast spots, with the affected area less than 10%

[0086] Level 7: Typical rice blast spots, with an affected area of ​​26%-50%

[0087] Level 9: Total leaf death

[0088] Calculation formula for biological control effect:

[0089]

[0090] Survey data

[0091]

[0092] Results: The color change of enzyme rice was more uniform than that of the control group, the disease was significantly less than that of the control group, the yield was higher than that of the control group, and the lodging resistance was significantly higher than that of the control group.

[0093] Pesticide residue detection in rice:

[0094] Drug testing method reference;

[0095] GB 23200.9-2016 National Food Safety Standard Determination of 475 Drugs and Related Chemical Residues in Cereals by Gas Chromatography-Mass Spectrometry and GB / T 20770-2008 Determination of 486 Drugs and Related Chemical Residues in Cereals by Liquid Chromatography-Tandem Mass Spectrometry;

[0096] Test table for rice sprayed with environmentally friendly enzymes:

[0097]

[0098]

[0099] Test table for conventionally grown rice:

[0100]

[0101] The total of 441 pesticide residue tests include 666, atrazine, chlormequat, aldrin, anthracene, chlorpyrifos, carbendazim and other routine 441 tests.

[0102] summary:

[0103] Taking rice as the research object, rice fields of the same variety in different regions were selected as the control group and experimental group for experiments. The control group was cultivated with conventional farming methods, and the experimental group was sprayed with enzymes. The effects of different process enzymes on loquat yield and quality were examined, and the differences in production performance indicators and rice pesticide residue-related indicators between the control group and the experimental group were analyzed. The pesticide residue detection results of the experimental groups all met the requirements. The rice appearance experiment showed that the experimental group was more complete, substantial, and fuller than the control group, and the taste was also better, and the experts gave good evaluations. The pesticide residue detection of the control group showed that the residues of chlorpyrifos and carbendazim exceeded the detection limit of the method. The color change of enzyme rice was more uniform than that of the control group, the disease was significantly lower than that of the control group, the yield was higher than that of the control group, and the lodging resistance was significantly higher than that of the control group.

[0104] IV: Conclusion of the test on the inhibition of loquat environmental enzyme on pathogenic bacteria

[0105] 1. Antibacterial effect of loquat enzyme on Candida albicans

[0106] The antibacterial activity of naturally fermented loquat enzyme against Candida albicans was investigated. The results showed that loquat enzyme had a good inhibitory effect on Candida albicans. The higher the enzyme content, the higher the antibacterial activity. The antibacterial activity of the enzyme at a concentration of 50% was as high as over 93%. Figure 2 .

[0107] 2. Antibacterial effect of loquat enzyme on Malassezia furfur

[0108] The antibacterial activity of naturally fermented loquat enzyme against Malassezia furfur was investigated. The results showed that loquat enzyme had a good inhibitory effect on Malassezia furfur. The inhibitory effect of low-concentration enzyme could reach more than 50%. The higher the enzyme content, the better the inhibitory effect. Figure 3 .

[0109] 3. Antibacterial effect of loquat enzyme on Bacillus aeruginosa

[0110] The antibacterial activity of naturally fermented loquat enzyme against Bacillus aeruginosa was investigated. The results showed that loquat enzyme had a good antibacterial effect. The antibacterial rate of 10% enzyme against Bacillus aeruginosa could reach more than 63%. When the enzyme concentration was higher than 25%, it completely inhibited the activity of Bacillus aeruginosa. Figure 4 .

[0111] 4. Antibacterial effect of loquat enzyme on Staphylococcus aureus

[0112] The antibacterial activity of naturally fermented loquat enzyme against Staphylococcus aureus was investigated. The results showed that loquat enzyme had a certain inhibitory effect on Staphylococcus aureus, and high concentration enzyme had a good antibacterial effect. Figure 5 .

[0113] Conclusion: Loquat enzyme has a certain inhibitory effect on both fungi and bacteria. Its mechanism of action may be that probiotics as dominant bacteria inhibit the growth of harmful bacteria, or it may be that the enzyme system (enzyme) secreted by fruits and vegetables themselves or microorganisms in the slow fermentation process has a decomposition and destruction effect on the cell wall of bacteria / fungi. Its effect is different from that of chemical antibiotics. It is not only effective against bacteria, but also against fungi. It strongly supports the conclusion of the above planting experiment, that is, environmentally friendly enzymes can replace various antibiotics in the planting process.

[0114] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0115] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. An application of an environmentally friendly enzyme for fruits and vegetables, characterized in that: The application of environmentally friendly fruit and vegetable enzymes in improving the growth performance of fruit trees and crops, specifically the use of environmentally friendly fruit and vegetable enzymes to replace chemical fertilizers and pesticides.

2. The method for preparing the environmentally friendly enzyme for fruits and vegetables according to claim 1, characterized in that: The specific steps are as follows: (1) Material selection: Select fruits and vegetables of the current season, clean them and dry them; (2) Environmental disinfection: spray the environment with a solution of Chlorhexidine and use ultraviolet light to disinfect for 15-60 minutes; (3) Equipment disinfection: Disinfect the fermentation tank with 70-75% alcohol as a disinfectant; (4) Culture medium formula: 7%-15% brown sugar, 20%-40% fruits and vegetables, 50%-73% water, pH is neutral, and the culture medium is prepared; (5) Production and feeding: Crush the fruits and vegetables and put them into the fermentation tank, the final volume of which shall not exceed 75%-80% of the space of the fermentation tank, package them, and label them. The label shall have the production date; (6) Directed fermentation: Use the dominant bacteria of fruits and vegetables to transform microorganisms and ferment naturally in a cool place for 2-4 months; (7) Release the tank: After the fermentation time expires, the fermentation liquid is clear and the fruits and vegetables are decomposed and settled, and the tank can be released. When releasing the fermentation tank, open the sampling port valve of the fermentation tank and take samples for inspection; (8) Test items: Take enzyme samples to test their properties and sensory indicators, general physical and chemical indicators of pH and TDS, and characteristic physical and chemical indicators of total acid, organic acid, and total bacterial count; (9) Solid-liquid separation: The fermentation liquid that meets the inspection standards is filtered through a 40-80 mesh screen or centrifuged in a box centrifuge to obtain the liquid enzyme product.

3. The method for producing an environmentally friendly fruit and vegetable enzyme according to claim 2, characterized in that: The fruits and vegetables in step (1) are seasonal fruit and vegetable waste collected from the market, with rotten and spoiled fruits and vegetables removed.

4. The method for producing an environmentally friendly enzyme for fruits and vegetables according to claim 2, characterized in that: In the step (6), exhaust is performed once every 3-4 days within half a month after the start of fermentation.

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