Method for producing enzyme fertilizer by taking perilla straw as raw material

Abstract

The invention discloses a production method of a ferment fertilizer by taking perilla straws as a raw material, which comprises the following steps: collecting the perilla straws and screening out unnecessary perilla leaves, crushing, sterilizing, adding soybean meal, and spraying a composite enzymatic hydrolysate for enzymolysis; and adding brown sugar and water, and carrying out aerobic fermentation and anaerobic fermentation. The purple perilla straw, the soybean meal and the like are subjected to enzymolysis and then are fermented, so that the fermentation speed can be increased, the utilization rate of resources such as the straw and the yield of fermentation products are increased, and the nutritional ingredients of the enzyme fertilizer are enriched.

Description

Technical Field

[0001] The invention relates to the technical field of enzyme fertilizers, and in particular to a method for producing enzyme fertilizers using perilla straw as a raw material. Background Art

[0002] Perilla has multiple uses, including food, medicine, and oil. Its leaves are rich in perilla aldehyde, perilla ketone, baicalin, menthol, alpine baicalin, New Zealand vitexin II, and scutellarin. Studies have shown that perilla aldehyde and other perilla extracts promote the growth of various plants and inhibit the growth of pathogens. In recent years, the perilla industry has shown steady growth and holds broad market prospects. However, most perilla straw is currently discarded or directly crushed and returned to the fields. A small amount is properly processed and used as livestock feed, and discarded perilla straw generally still contains a small amount of fresh leaves, which prevents its high-value development and utilization.

[0003] Enzyme microbial fertilizer, a type of fertilizer rich in beneficial bacteria and trace elements, is produced through fermentation using active microorganisms. Due to its environmentally friendly and efficient properties, it is becoming a highly regarded new fertilizer in modern agriculture. However, because perilla straw primarily consists of cellulose and hemicellulose (it also contains small amounts of volatile oils, flavonoids, and phenolic acids), existing liquid enzyme fertilizer production methods result in a slow fermentation temperature increase and difficulty reaching temperatures above 50°C. The resulting product is low quality and prone to carrying pathogens. Summary of the Invention

[0004] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a method for producing enzyme fertilizer using perilla straw as raw material.

[0005] To achieve the above object, the technical solution adopted by the present invention is as follows: A method for producing enzyme fertilizer using perilla straw as raw material, characterized by comprising the following steps: Step 1) collecting the perilla stalks and the discarded perilla leaves, crushing and sterilizing them to obtain perilla powder; Step 2) Add soybean meal to the crushed perilla powder, spray the composite enzymatic hydrolysate, mix well and perform enzymatic hydrolysis for 4-6 hours to obtain a solid-liquid mixture, stirring regularly during the enzymatic hydrolysis process; Step 3) First, brown sugar and water are added to the solid-liquid mixture obtained in step 2), and then Bacillus subtilis is inoculated to carry out the first stage of aerobic fermentation. After the first stage of aerobic fermentation is completed, yeast is inoculated to continue aerobic fermentation for 5-10 days; then lactic acid bacteria are inoculated to carry out the second stage of anaerobic fermentation; Step 4): After the fermentation process in step 3), solid-liquid separation is performed, wherein the liquid portion is the target liquid enzyme fertilizer.

[0006] Furthermore, the perilla straw in step 1) above is fresh straw of perilla aldehyde type, perilla ketone type and hyssostigmine type.

[0007] Furthermore, the sterilization treatment in the above step 1) is specifically to soak the crushed perilla straw in a proper amount of tap water for 1-3 hours, then add a proper amount of water to boil for 0.5-1.5 hours and then drain.

[0008] Furthermore, the perilla straw in the above step 1) is crushed into 3-5 cm long straw powder.

[0009] Furthermore, in the above step 2), the amount of the composite enzymatic hydrolysis solution is 8%-15% of the total weight of the perilla powder, and the amount of soybean meal is 10-20% of the total weight of the perilla powder, and the composite enzymatic hydrolysis solution is obtained by dissolving the composite enzyme in a phosphate buffer solution, wherein the composite enzyme is composed of cellulase, ligninase, xylanase, and protease in a weight ratio of 2-4:1-3:1-2:1-2, and the mass ratio of the composite enzyme to the phosphate buffer solution is 10-15:1.

[0010] Furthermore, in the above step 3), the amount of brown sugar and water added is 30%-50% and 2-5 times of the chopped perilla powder, respectively.

[0011] Furthermore, in the above step 3), the dosage of Bacillus aspartate, yeast, and lactic acid bacteria is 8%-10% of the total weight of the crushed perilla powder.

[0012] Furthermore, in the above step 3), the time of the first stage aerobic fermentation is controlled to be 0.8-1.2 months; and the time of the second stage anaerobic fermentation is controlled to be 1.5-2.5 months.

[0013] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention first enzymatically hydrolyzes perilla straw and soybean meal and then ferments them, which can accelerate the fermentation speed, improve the utilization rate of resources such as straw and the yield of fermentation products, and enrich the nutritional components of enzyme fertilizer; 2. The present invention adopts a combination of aerobic fermentation and anaerobic fermentation, which can make the fermentation more thorough, enrich the biological activity and diversity of the liquid enzyme fertilizer product, and at the same time reduce the accumulation of harmful substances and improve the safety and quality of the liquid enzyme fertilizer; 3. The present invention sterilizes the straw before enzymatic hydrolysis and combines enzymatic hydrolysis, aerobic and anaerobic fermentation, which can effectively reduce the competition and production inhibition of miscellaneous bacteria and impurities during the enzymatic hydrolysis and fermentation process, thereby reducing production costs;

[0014] 4. The liquid enzyme fertilizer prepared by the present invention has the functions of improving soil and promoting plant growth, and is suitable for organic agriculture and green agriculture. DETAILED DESCRIPTION

[0015] Now, in conjunction with specific embodiments, the present invention will be further described. The following embodiments are only preferred embodiments of the present invention and do not limit the scope of the present invention. Any scheme without departing from the concept of the present invention should fall within the scope of protection of the present invention.

[0016] Example 1 (JS1) The present embodiment uses the enzyme fertilizer production method of perilla straw as raw material, and the specific steps are as follows: Step 1) collecting different types of dried perilla stalks and the discarded dried perilla leaves, first crushing them into 3-5 cm long straw powders, then soaking the crushed perilla stalks in appropriate amount of tap water for 3 hours, then boiling them in appropriate amount of water for 0.5 hours, draining them, and cooling them to below 60° C. to obtain perilla powder; Step 2), adding soybean meal to the perilla powder, the amount of soybean meal is 20% of the total weight of the perilla powder, and spraying the composite enzymatic hydrolysis solution, the amount of the composite enzymatic hydrolysis solution is 15% of the total weight of the perilla powder, and the composite enzymatic hydrolysis solution is obtained by dissolving the composite enzyme in a phosphate buffer solution, wherein the composite enzyme is composed of cellulase (compounded by endoglucanase, exoglucanase, and β-glucosidase in a weight ratio of 1:1:1), ligninase (compounded by laccase and manganese peroxidase in a weight ratio of 1:1), xylanase (compounded by endoxylanase and exoxylanase in a weight ratio of 1:1), protease (compounded by neutral protease and papain in a weight ratio of 1:1) in a weight ratio of 4:1:1:1, and the mass ratio of the composite enzyme to the phosphate buffer solution is 10:1; mixing and enzymolysis for 4 hours to obtain a solid-liquid mixture, and stirring once every 10 minutes during the enzymolysis process; Step 3) First, brown sugar and water are added to the solid-liquid mixture obtained by the treatment in step 2), with the amount of brown sugar and water added being 50% and 5 times the weight of the crushed perilla powder, respectively; then, Bacillus subtilis is inoculated to carry out a first-stage aerobic fermentation for a controlled period of 0.8 months; after the first-stage aerobic fermentation is completed, yeast is inoculated to continue aerobic fermentation for 5 days; then, lactic acid bacteria is inoculated to carry out a second-stage anaerobic fermentation for a controlled period of 1.5 months; the amount of Bacillus aspartate, yeast, and lactic acid bacteria is 10% of the total weight of the crushed perilla powder; Step 4): After the fermentation process in step 3), solid-liquid separation is performed, wherein the liquid part is liquid enzyme fertilizer.

[0017] Example 2 (JS2) The present embodiment uses the enzyme fertilizer production method of perilla straw as raw material, and the specific steps are as follows: Step 1), selecting fresh perilla aldehyde-type perilla straw and the selected fresh leaves, first crushing them into 3-5 cm long straw powder, then adding appropriate amount of tap water to soak the crushed perilla straw for 1 hour, then adding appropriate amount of water to boil for 1.5 hours, draining, and cooling to below 60°C to obtain perilla powder; Step 2), adding soybean meal to the perilla powder, the amount of soybean meal is 10% of the total weight of the perilla powder, and spraying the composite enzymatic hydrolysis solution, the amount of the composite enzymatic hydrolysis solution is 8% of the total weight of the perilla powder, and the composite enzymatic hydrolysis solution is obtained by dissolving the composite enzyme in a phosphate buffer solution, wherein the composite enzyme is composed of cellulase (compounded by endoglucanase, exoglucanase, and β-glucosidase in a weight ratio of 1:1:1), ligninase (compounded by laccase and manganese peroxidase in a weight ratio of 1:1), xylanase (compounded by endoxylanase and exoxylanase in a weight ratio of 1:1), protease (compounded by neutral protease and papain in a weight ratio of 1:1) in a weight ratio of 4:2:2:1, and the mass ratio of the composite enzyme to the phosphate buffer solution is 15:1; mixing and enzymolysis for 6 hours to obtain a solid-liquid mixture, and stirring once every 20 minutes during the enzymolysis process; Step 3) First, brown sugar and water are added to the solid-liquid mixture obtained by the treatment in step 2), with the amount of brown sugar and water being 35% and 3 times the weight of the chopped perilla powder, respectively; then, Bacillus subtilis is inoculated to carry out a first-stage aerobic fermentation for a controlled period of 1.2 months. After the first-stage aerobic fermentation is completed, yeast is inoculated to continue aerobic fermentation for 10 days; then, lactic acid bacteria is inoculated to carry out a second-stage anaerobic fermentation for a controlled period of 2.5 months; the amount of Bacillus aspartate, yeast, and lactic acid bacteria is 8% of the total weight of the chopped perilla powder; Step 4): After the fermentation process in step 3), solid-liquid separation is performed, wherein the liquid part is liquid enzyme fertilizer.

[0018] Example 3 (JS3) Except for the following differences, the rest is the same as Example 2: The fresh straw of perilla ketone type perilla and the unnecessary fresh leaves that have been screened out are selected.

[0019] Example 4 (JS4) Except for the following differences, the rest is the same as Example 2: The fresh straw of the oleic acid type perilla and the unnecessary fresh leaves that have been screened out are selected.

[0020] A field experiment was conducted with cotton as the experimental object. Five treatment groups were set up (see Table 1 for details). Each group had three replicate plots, and each plot was 30 m 2 (1.5×20), the experiment adopted a randomized block design.

[0021] Table 1 Specific implementation plans for different treatments

[0022] During the experiment, the growth period, agronomic characteristics of the plants, physiological and biochemical indicators, etc. were investigated, tested and recorded.

[0023] Five days after sowing, cotton seedling emergence was monitored and recorded daily. Fertilization began once the seedlings were fully grown, and cotton growth was monitored and recorded every ten days. Table 2 shows that application of perilla enzyme fertilizers can advance the bud and boll formation stages of cotton, with the scutellarin-based perilla enzyme fertilizer (JS4) showing the most significant effect.

[0024] Table 2 Effects of different treatments on cotton growth period

[0025] The plant height, leaf area and other traits of cotton were measured and recorded after fertilization for 2 times and 5 times respectively. As shown in Table 3, after fertilization for 2 times and 5 times, the plant height growth value and leaf area of cotton in the treatment group treated with perilla enzyme fertilizer were higher than those in the control group. Among them, the enzyme fertilizer (JS2) produced from fresh perilla straw of perilla aldehyde type had the strongest promoting effect on the growth of cotton seedlings after fertilization for 2 times; the enzyme fertilizer (JS4) produced from fresh perilla straw of chrysanthene type had the strongest promoting effect on the growth of cotton seedlings after fertilization for 5 times.

[0026] Table 3 Effects of different treatments on cotton seedling growth

[0027] Physiological and biochemical indicators such as cotton leaf chlorophyll content, nitrogen content, and photosynthetic rate were measured and recorded after two and five fertilizations. As shown in Table 4, except for the JS1 and JS3 treatment groups, which had lower chlorophyll content in cotton leaves than the control group after two fertilizations, the chlorophyll content and nitrogen content in the leaves of the other treatment groups were higher than those of the control group after two and five fertilizations, with the JS4 treatment group having the highest chlorophyll content. The photosynthetic rate of cotton in the JS1 and JS4 treatment groups was slightly lower than that of the control group, while the photosynthetic rate of cotton in the JS2 and JS3 treatment groups was significantly higher than that of the control group. This indicates that the application of perilla enzyme fertilizer can increase the chlorophyll and nitrogen content in cotton leaves, and that enzyme fertilizers produced from perilla aldehyde and perilla ketone fresh perilla straw can effectively increase the photosynthetic rate of cotton.

[0028] Table 4 Effects of different treatments on cotton physiology and biochemistry

[0029] After five fertilizations, cotton rhizosphere soil was collected to measure rhizosphere microbial diversity. Table 5 shows that the Shannon index of each enzyme fertilizer treatment group was higher than that of the control group. At the bacterial phylum level, the relative abundance of Firmicutes, Actinobacteria, and Proteobacteria in each enzyme fertilizer treatment group was higher than that of the control group. Numerous studies have shown that these three bacterial phyla, including rhizosphere growth-promoting bacteria such as Streptomyces, Nocardia, Rhizobia, and Bacillus, promote plant growth. This suggests that the application of perilla enzyme fertilizer can increase cotton rhizosphere microbial diversity and promote the proliferation of rhizosphere growth-promoting bacteria.

[0030] Table 5 Effects of different treatments on cotton soil microorganisms

[0031] In summary, enzyme fertilizer produced from waste perilla straw has a promoting effect on cotton growth and improves the rhizosphere microbial structure. Based on the experimental results, enzyme fertilizers produced from perilla ketone and perilla aldehyde types of perilla straw are more effective in promoting cotton growth and improving rhizosphere microbial structure.

Claims

1. A method for producing enzyme fertilizer using perilla straw as raw material, characterized in that: The steps include: Step 1) collecting the perilla stalks and the discarded perilla leaves, crushing and sterilizing them to obtain perilla powder; Step 2) Add soybean meal to the crushed perilla powder, spray the composite enzymatic hydrolysate, mix well and perform enzymatic hydrolysis for 4-6 hours to obtain a solid-liquid mixture, stirring regularly during the enzymatic hydrolysis process; Step 3) First, brown sugar and water are added to the solid-liquid mixture obtained in step 2), and then Bacillus subtilis is inoculated to carry out the first stage of aerobic fermentation. After the first stage of aerobic fermentation is completed, yeast is inoculated to continue aerobic fermentation for 5-10 days; then lactic acid bacteria are inoculated to carry out the second stage of anaerobic fermentation; Step 4): After the fermentation process in step 3), solid-liquid separation is performed, wherein the liquid part is liquid enzyme fertilizer.

2. A method for producing enzyme fertilizer using perilla straw as raw material according to claim 1, characterized in that: The perilla straw in step 1) above is fresh straw of perilla aldehyde type, perilla ketone type and hyssostigmine type.

3. A method for producing enzyme fertilizer using perilla straw as raw material according to claim 2, characterized in that: The sterilization treatment in the above step 1) is specifically to soak the crushed perilla straw in a proper amount of tap water for 1-3 hours, then add a proper amount of water to boil for 0.5-1.5 hours and then drain.

4. A method for producing enzyme fertilizer using perilla straw as raw material according to claim 2, characterized in that: In step 1), the perilla stalks are crushed into 3-5 cm long pieces.

5. A method for producing an enzyme fertilizer using perilla straw as raw material according to claim 1, 2, 3 or 4, wherein: In the above step 2), the amount of the composite enzymatic hydrolysis solution is 8%-15% of the total weight of the perilla powder, and the amount of soybean meal is 10-20% of the total weight of the perilla powder. The composite enzymatic hydrolysis solution is obtained by dissolving a composite enzyme in a phosphate buffer solution, wherein the composite enzyme is composed of cellulase, ligninase, xylanase, and protease in a weight ratio of 2-4:1-3:1-2:1-2, and the mass ratio of the composite enzyme to the phosphate buffer solution is 10-15:

1.

6. A method for producing enzyme fertilizer using perilla straw as raw material according to claim 5, characterized in that: In the above step 3), the amount of brown sugar and water added is 30%-50% and 2-5 times of the chopped perilla powder respectively.

7. A method for producing enzyme fertilizer using perilla straw as raw material according to claim 6, characterized in that: In the above step 3), the amount of Bacillus aspartate, yeast, and lactic acid bacteria is 8%-10% of the total weight of the crushed perilla powder.

8. A method for producing enzyme fertilizer using perilla straw as raw material according to claim 7, characterized in that: In the above step 3), the first stage aerobic fermentation time is controlled to be 0.8-1.2 months; the second stage anaerobic fermentation time is controlled to be 1.5-2.5 months.

Images