Modified plant extract for degrading ammonia gas as well as preparation method and application of modified plant extract

Abstract

The invention discloses a modified plant extract for degrading ammonia gas as well as a preparation method and application thereof, and belongs to the technical field of deodorants. Comprising the following steps: (1) dissolving a rapeseed extract, and adding glyceraldehyde to obtain a rapeseed extract-glyceraldehyde mixed solution; and (2) carrying out grafting reaction on the rapeseed extract-glyceraldehyde mixed solution, centrifuging and dialyzing to obtain the modified plant extract. Compared with a traditional enzyme deodorant method, the method has the advantages that the method for modifying the selected plant extract is easy to implement, simple, safe and convenient for experimental large-scale production, the deodorization rate for ammonia gas is high, about 85% of ammonia gas can be degraded, and the plant extract is modified through a damp-heat method, so that the production efficiency is high.

Description

Technical Field

[0001] The invention relates to the technical field of deodorants, and in particular to a modified plant extract for degrading ammonia, and a preparation method and application thereof. Background Art

[0002] Lipase belongs to the carboxyl ester hydrolase class and can gradually hydrolyze triglycerides into glycerol and fatty acids. Lipase exists in fat-containing animal, plant and microbial (such as mold, bacteria, etc.) tissues, including phosphatase, sterolase and carboxylesterase. Fatty acids are widely used in food, medicine, leather, daily chemicals and other aspects.

[0003] Lipase is widely present in animals, plants and microorganisms. The seeds of oil crops, such as castor seeds and rapeseed, contain more lipase in plants. When the oil seeds germinate, lipase can work together with other enzymes to catalyze the decomposition of oil substances to produce sugars, providing the nutrients and energy necessary for the rooting and germination of seeds; the pancreas and adipose tissue of higher animals contain more lipase in animals. There is a small amount of lipase in the intestinal fluid to supplement the insufficient fat digestion of pancreatic lipase. There is a small amount of butyric acid glycerol esterase in the gastric juice of carnivorous animals. In animals, various types of lipase control processes such as digestion, absorption, fat reconstruction and lipoprotein metabolism; the lipase content in bacteria, fungi and yeast is richer. Due to the large variety of microorganisms, fast reproduction and easy genetic variation, they have a wider range of pH, temperature and substrate specificity than animals and plants, and lipases from microorganisms are generally secreted extracellular enzymes. The main fermentation microorganisms are Aspergillus niger, Candida, etc. It is suitable for large-scale industrial production and obtaining high-purity samples. Therefore, microbial lipase is an important source of industrial lipase. Generally, lipases from different sources have different characteristics and are also of great significance in theoretical research.

[0004] Glyceraldehyde is an organic compound with a molecular formula of C3H6O3 and a molecular weight of 90.08. It is a triose (monosaccharide) and the simplest aldose. It is a sweet white crystal and an intermediate product of carbohydrate metabolism. Glyceraldehyde and dihydroxyacetone can be prepared by mild oxidation of glycerol. The oxidant can be hydrogen peroxide, catalyzed by a divalent iron salt. L-sorbose can be oxidized and broken to obtain L-glyceraldehyde. D-fructose can be oxidized and broken to obtain D-glyceraldehyde. Glyceraldehyde is easily converted into dihydroxyacetone by enolization, and forms an equilibrium mixture with the latter in aqueous solution, called triose. Its aldehyde group is relatively active and is generally stored in the form of methanol acetal.

[0005] The article Identification, purification, and characterization of a thermallystable lipase from rice bran. A new member of the (phospho) lipase family (DOI: 10.1104 / PP.010604) first discovered a thermallystable lipase (EC3.1.1.3.) in rice (Oryza sativa) bran and purified the enzyme to homogeneity using octyl agarose chromatography. Using [9,10-(3)H]triolein as a substrate, the enzyme was purified 7.6 times at 80 °C, with a final specific activity of 0.38 μmol•min ( -1 )•mg( -1 ). The article Purification and Characterization, of Rice Bran Lipase II (DOI: 10.1080 / 00021369.1976.10862049) purified a rice bran lipase (lipase II) by ammonium sulfate precipitation, followed by sequential chromatography on DEAE-cellulose, Sephadex G-75, and CH-Sephadex C-50. Both polyacrylamide disk electrophoresis and ultracentrifugation showed that the enzyme protein was homogeneous.

[0006] The use of polymers to modify lipases further broadens the application scope of polymer-modified enzymes. At the same time, there are a wide variety of polymers available for modification, providing a variety of options. Therefore, this research field has broad prospects.

[0007] Ammonia is a colorless gas with a strong pungent odor. It is mostly found in industrial waste gas, livestock breeding factories, sewers, etc. At present, the commonly used methods at home and abroad are adsorption method, combustion method, high-energy ion method and biological method. At present, for the treatment of malodorous gases such as ammonia, it is necessary to find a low-cost, high-efficiency and widely applicable method. Summary of the invention

[0008] The purpose of the present invention is to provide a modified plant extract for degrading ammonia and a preparation method and application thereof, so as to solve the problems existing in the above-mentioned prior art.

[0009] To achieve the above object, the present invention provides the following solutions: One of the technical solutions of the present invention is a method for preparing a modified plant extract for degrading ammonia, comprising the following steps: (1) dissolving the rapeseed extract and adding glyceraldehyde to obtain a rapeseed extract-glyceraldehyde mixed solution; (2) subjecting the rapeseed extract-glyceraldehyde mixed solution to a grafting reaction, centrifuging, and dialyzing to obtain the modified plant extract.

[0010] The second technical solution of the present invention is the modified plant extract prepared by the preparation method.

[0011] The third technical solution of the present invention is the use of the modified plant extract in the degradation of ammonia.

[0012] A fourth technical solution of the present invention is a deodorant for degrading ammonia, comprising the modified plant extract.

[0013] Based on the above technical solution, the present invention has the following technical effects: Compared with the traditional enzyme deodorant method, the advantages of the method of the present invention are mainly reflected in the following aspects: 1) The modification method of the selected plant extracts is easy to implement, simple and safe, and convenient for experimental large-scale production; 2) The deodorization rate of ammonia is high, and it can degrade about 85% of ammonia; 3) Modification of plant extracts by wet heat method has high production efficiency and is green and environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0015] Figure 1 The figure is a flow chart of the experiment of the embodiment of the present invention.

[0016] Figure 2 This is a diagram of the identification results of the rapeseed extract prepared by the present invention. The samples from the left to right are: rapeseed oil standard (ethyl acetate), rapeseed oil standard (n-butanol), 90min sample, 120min sample, 160min sample, 220min sample, 260min sample, and 300min sample. DETAILED DESCRIPTION

[0017] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0018] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0019] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0020] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0021] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0022] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0023] The embodiment of the present invention provides a method for preparing a modified plant extract for degrading ammonia, comprising the following steps: (1) dissolving the rapeseed extract and adding glyceraldehyde to obtain a rapeseed extract-glyceraldehyde mixed solution; (2) subjecting the rapeseed extract-glyceraldehyde mixed solution to a grafting reaction, centrifuging, and dialyzing to obtain the modified plant extract.

[0024] In some specific embodiments, the solvent for dissolving the rapeseed extract is PBS buffer with a pH of 4-11, preferably 8.

[0025] In some specific embodiments, the mass ratio of the rapeseed extract to glyceraldehyde is 1:(1-10), preferably 1:5.

[0026] In some specific embodiments, the grafting reaction conditions are: heating in a water bath at 30-60° C. for 30-90 min, preferably heating in a water bath at 45° C. for 50 min.

[0027] In some specific embodiments, the centrifugation condition is: centrifugation at 6000 r / min for 5 min.

[0028] In some specific embodiments, the dialysis condition is: dialysis in distilled water at 4°C for 20 hours.

[0029] The embodiment of the present invention also provides a modified plant extract prepared by the preparation method.

[0030] The embodiment of the present invention also provides the use of the modified plant extract in degrading ammonia.

[0031] The embodiment of the present invention also provides a deodorant for degrading ammonia, comprising the modified plant extract.

[0032] The problem to be solved by the present invention is to prepare a modified plant extract, conduct a deodorization experiment, and make a deodorant of a natural extract. The basic concept of the technical solution adopted by the present invention is: accurately weigh a certain mass of rapeseed plant extract according to a proportion, add it to a PBS solution with a certain pH value, stir and dissolve it, and then put it in a refrigerator for storage. Accurately take a certain mass of glyceraldehyde according to a proportion, add it to the PBS mixed solution in the first step, stir and dissolve it, and then put it in a refrigerator for storage. The rapeseed plant extract is modified by a wet heat method to obtain a rapeseed extract-glyceraldehyde grafted product, and the grafted product is dialyzed, freeze-dried, and powdered. By adjusting the wet heat process temperature, pH value, time and other conditions, a grafted product is obtained, and the product can degrade ammonia in the air and achieve a deodorizing effect.

[0033] According to the reaction flow chart, the preparation process is: accurately weigh, stir and dissolve, put into a test tube for hydrothermal treatment, adjust the hydrothermal temperature, take out after an appropriate hydrothermal time, put in an ice water bath to end the experiment, centrifuge the mixture to retain the supernatant, dialyze the supernatant against 4°C distilled water, and freeze-dry the dialyzate to obtain the rapeseed extract-glyceraldehyde grafted product.

[0034] Example 1 Preparation of rapeseed extract: 40g of rapeseed seeds were processed with a pulverizer at 3800r / min for 10min, and 20mL of 4°C acetone was added thereto. The obtained paste was stirred at a constant temperature of 4°C (700rpm) for 10h. Filtrate 1 and solid substance 1 were then obtained by vacuum filtration. In order to improve the extraction rate, solid substance 1 was washed several times with 4°C acetone to separate filtrate 2. The obtained filtrate 1 and filtrate 2 were allowed to stand in a sterile environment at room temperature until completely dried, and a solid powder was obtained after the solvent was completely evaporated, which was the rapeseed extract. The solid powder was passed through a 40-mesh sieve, bottled and sterilized at 4°C for later use.

[0035] Identification of Rapeseed Extract: Thin layer chromatography was used with rapeseed oil as the substrate. 50 ml of rapeseed oil was taken, 10 g of "rapeseed extract" was added, and the reaction was shaken at a constant temperature of 38°C. Sampling was performed at 90, 120, 160, 220, 260, and 300 min. Ethyl acetate was used as the extractant, and 100 μL of sample was added to each milliliter of ethyl acetate. Ethyl acetate and n-butanol were used as the rapeseed oil extractants (to explore which extractant was more suitable), respectively. The extractants were shaken at high speed until uniform and set aside. The developing agent ratio was chloroform: methanol: water = 7:3:1, and the color developer was 10% ethanolic sulfuric acid solution. The capillary glass tube was used to spot the plate, and the plate was placed in the extraction cylinder after it was completely dry. After the extraction was completed, it was dried in an oven at 80°C and placed on a color developer at 120°C for color development.

[0036] The results are as follows Figure 2 As shown, the TLC results proved that the acetone extract contained rapeseed lipase, which could decompose rapeseed oil.

[0037] Ammonia degradation test: According to the national standard HJ 533-2009, the degradation rate of rapeseed extract to ammonia was tested by Nessler reagent spectrophotometer method. 5 grams of modified plant extract was placed in a container and 1 m 3 ammonia, connect the container to a conical flask containing 50ml sulfuric acid absorption liquid (prepared according to national standards) through a glass tube, seal the bottle mouth with a balloon to ensure that the conical flask container is airtight, and collect the gas for 40 minutes to obtain the sample to be tested; take 0.1ml of the sample to be tested and add it to a colorimetric tube, add 4.9ml double distilled water, add 0.1ml potassium sodium tartrate solution (CAS: 304-59-6, purchased from Shanghai Titan Technology Co., Ltd.), 0.1ml Nessler's reagent (purchased from Shanghai Titan Technology Co., Ltd.), let it stand for 10 minutes, and use an ultraviolet spectrophotometer to test the absorbance at 420nm. The blank group is a water sample, and the data is collected to calculate the ammonia degradation rate.

[0038] The control group did not add plant extracts, 1m 3 Ammonia gas is passed into 50 ml of sulfuric acid absorption liquid, and the control group data is x2: the test concentration of undegraded ammonia gas.

[0039] Weigh 4 grams of homemade rapeseed extract from the laboratory accurately according to the proportion, add it to 100 mL of PBS buffer solution with a pH of 8, stir with a magnetic stirrer for 40 minutes, and store the solution in a 4°C refrigerator after it is fully dissolved; weigh 20 grams of glyceraldehyde accurately according to the proportion, add glyceraldehyde to the rapeseed extract-PBS buffer solution and stir for 15 minutes, so that it is fully fused and stored in a 4°C refrigerator for later use. Take 25 mL of the rapeseed extract-glyceraldehyde mixture and put it in a stoppered test tube, place it in a water bath at 45°C for grafting reaction for 50 minutes, and immediately put it in an ice water bath after the water bath ends to end the reaction. After centrifugation at 6000r / min for 5 minutes, take the supernatant and dialyze it with 4°C distilled water for 20 hours. Precool the dialysate in a -20°C refrigerator, then freeze-dry it in a freeze dryer for 24 hours and grind it into powder to obtain the modified rapeseed extract. Put 5 grams of modified plant extract in a container and pass 1m 3 ammonia, and after 60 minutes, the ammonia content in the container was detected to obtain the ammonia degradation rate. The calculation formula is as follows: y=0.0054x+0.0147, R²=0.9921; Where, y: absorbance, x: sample ammonia concentration.

[0040] Ammonia degradation rate Y = (x2-x1) / x2*100%; Wherein, Y is the ammonia degradation rate; x1 is the test concentration of ammonia after degradation in the container; x2 is the test concentration of undegraded ammonia.

[0041] Example 2 The only difference from Example 1 is that the pH of the PBS buffer solution is 4, and the remaining steps are completely consistent with Example 1.

[0042] Example 3 The only difference from Example 1 is that the pH of the PBS buffer solution is 6, and the remaining steps are completely consistent with Example 1.

[0043] Example 4 The only difference from Example 1 is that the pH of the PBS buffer solution is 11, and the remaining steps are completely consistent with Example 1.

[0044] Example 5 The only difference from Example 1 is that the grafting reaction time is 30 minutes, and the other steps are completely consistent with Example 1.

[0045] Example 6 The only difference from Example 1 is that the grafting reaction time is 70 minutes, and the other steps are completely consistent with Example 1.

[0046] Example 7 The only difference from Example 1 is that the grafting reaction time is 90 minutes, and the other steps are completely consistent with Example 1.

[0047] Example 8 The only difference from Example 1 is that the mass ratio of rapeseed extract to glyceraldehyde is 1:1 (ie, the amount of glyceraldehyde is 4 g), and the remaining steps are completely consistent with Example 1.

[0048] Example 9 The only difference from Example 1 is that the mass ratio of rapeseed extract to glyceraldehyde is 1:2 (ie, the amount of glyceraldehyde is 8 g), and the remaining steps are completely consistent with Example 1.

[0049] Example 10 The only difference from Example 1 is that the mass ratio of rapeseed extract to glyceraldehyde is 1:10 (ie, the amount of glyceraldehyde is 40 g), and the remaining steps are completely consistent with Example 1.

[0050] Embodiment 11 The only difference from Example 1 is that the temperature of the grafting reaction is 30° C., and the other steps are completely consistent with Example 1.

[0051] Example 12 The only difference from Example 1 is that the temperature of the grafting reaction is 60° C., and the other steps are completely consistent with Example 1.

[0052] Results The ammonia degradation rate results of Examples 1-12 are shown in Table 1.

[0053] Table 1

[0054] It can be seen from Table 1 that the modification method of Example 1 of the present invention has the highest efficiency. By comparing Examples 1-12, it can be seen that the deodorization effect changes to varying degrees by changing the conditions of the modification scheme, including the pH value of the PBS buffer solution, the grafting time, the raw material ratio and the grafting temperature. It is meaningless to have a too long grafting time.

[0055] The present invention takes ammonia as the main deodorization research object, mixes rapeseed extract and glyceraldehyde in proportion, and grafts them by wet heat method. The modification of the extract is achieved by controlling the temperature, pH value, reaction time, etc. of the modification process. The modified extract is used to test the degradation of ammonia, and a modified plant extract that can deodorize is successfully developed.

[0056] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A method for preparing a modified plant extract for degrading ammonia, characterized in that: The following steps are involved: (1) dissolving the rapeseed extract and adding glyceraldehyde to obtain a rapeseed extract-glyceraldehyde mixed solution; (2) subjecting the rapeseed extract-glyceraldehyde mixed solution to a grafting reaction, centrifuging, and dialyzing to obtain the modified plant extract.

2. The preparation method according to claim 1, characterized in that: The solvent for dissolving the rapeseed extract is PBS buffer with a pH value of 4-11.

3. The preparation method according to claim 1, characterized in that: The mass ratio of the rapeseed extract to glyceraldehyde is 1:(1-10).

4. The preparation method according to claim 1, characterized in that: The grafting reaction conditions are: heating in a water bath at 30-60° C. for 30-90 min.

5. The preparation method according to claim 1, characterized in that: The centrifugal condition is: 6000r / min for 5min.

6. The preparation method according to claim 1, characterized in that: The dialysis conditions are: dialysis in distilled water at 4°C for 20 hours.

7. The modified plant extract prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the modified plant extract as claimed in claim 7 in degrading ammonia.

9. A deodorant for degrading ammonia, characterized in that: The modified plant extract according to claim 7.

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