CN107320451B - Lipase wettable powder and application thereof in silkworm breeding - Google Patents
Lipase wettable powder and application thereof in silkworm breeding Download PDFInfo
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- CN107320451B CN107320451B CN201710589580.6A CN201710589580A CN107320451B CN 107320451 B CN107320451 B CN 107320451B CN 201710589580 A CN201710589580 A CN 201710589580A CN 107320451 B CN107320451 B CN 107320451B
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- lipase
- wettable powder
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/465—Hydrolases (3) acting on ester bonds (3.1), e.g. lipases, ribonucleases
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/145—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/01—Carboxylic ester hydrolases (3.1.1)
- C12Y301/01003—Triacylglycerol lipase (3.1.1.3)
Abstract
The invention discloses lipase wettable powder and application thereof in silkworm breeding. The wettable powder comprises the following raw materials in percentage by mass: 75-85% of lipase raw powder, 5-12% of wetting agent, 5-12% of dispersing agent, 1-3% of stabilizing agent and 1-3% of protective agent. The lipase wettable powder disclosed by the invention can be used for silkworm breeding, and has the functions of promoting the growth and development of silkworms and improving the disease resistance and the yield of silkworm cocoons. The lipase wettable powder is diluted by adding water in proportion, and is applied to the leaves of oak, mulberry and the like by spraying, or is used for feeding tussah, mulberry and the like after directly soaking the leaves. The preparation method of the lipase wettable powder has the characteristics of simple process, low cost, easy industrial production and the like, and the preparation is safe, nontoxic and residue-free to the environment, people and livestock; simple application mode, obvious yield increasing effect, easy popularization and application and the like.
Description
Technical Field
The invention relates to lipase wettable powder and application thereof in silkworm breeding.
Background
Lipase (EC 3.1.1.3), triacylglycerol acyl hydrolase, is an enzyme that can catalyze the hydrolysis of long chain fatty glyceride and also the reverse reaction of the reaction, esterification. The lipase has important physiological functions in organisms, exogenous fat can permeate cell membranes after being digested and decomposed by the lipase, the storage and the utilization of the fat in the organisms are less than that of the lipase, and the lipase also participates in intracellular lipid metabolism. Monoacylglycerols, fatty acids, and glycerols produced by decomposition of triacylglycerols with lipases are precursors of lipids having important physiological functions, such as synthetic phospholipids and sphingolipids, in addition to being energy sources for living bodies. The lipase is not only beneficial to the digestion and absorption of exogenous fat, but also can release the fat contained in the feed raw materials, thereby meeting the requirements of animals on the fat. The lipase has the advantages of no pollution, mild reaction conditions and the like, and is widely applied to the traditional fields of washing, tanning, food, feed and the like.
At present, lipase has been approved by the nation as a feed additive, and about more than ten companies produce feed-grade lipase at home and abroad. By applying the lipase in the feed, the defect of lack of endogenous digestive enzymes of animals can be improved to a great extent, and the digestion of fat and the rapid growth of the animals are promoted, so that the application prospect of the lipase in the animal feed is certainly widened.
Silkworm is an economic insect with great economic and social benefits and has important application value in the industries of textile, food, medicine and the like. The silkworm variety includes silkworm, tussah silkworm, castor silkworm, cassava silkworm, coriaria sinica, Huili silkworm, Japanese tussah silkworm, amber silkworm, camphor silkworm, chestnut silkworm, Ailanthus silkworm, Chinese tallow tree silkworm, willow silkworm, etc. Most of products applied to silkworm breeding at present are silkworm feeds, the added substances are amino acids, vitamins, antibacterial agents and the like, the problem of low conversion rate (about 10%) of leaf silks always exists, and meanwhile, the loss amount of silkworm cocoons caused by silkworm diseases reaches over 10% every year, so that how to improve the conversion rate of the leaf silks and reduce the incidence rate of the silkworm diseases are important subjects faced by the silkworm industry production.
Disclosure of Invention
Aiming at the problems, the invention provides the lipase wettable powder, and the lipase wettable powder is innovatively applied to silkworm breeding, so that the growth and development of silkworms are obviously promoted, and the disease resistance and the silkworm cocoon yield of the silkworms are improved.
The inventor finds that the lipase is neutral or alkaline lipase, the enzyme activity is higher and more stable in an alkaline environment generally, while silkworms belong to insects, and different from a common animal digestive system, digestive juice of the silkworms is alkaline, so that the property of the lipase is suitable for being applied to the breeding of the silkworms, and the lipase has remarkable effects of improving the conversion rate of leaves in silkworm bodies, reducing the incidence rate of silkworm diseases, and improving the yield and quality of silk.
However, the feeding amount of the lipase by silkworms is a key problem for guaranteeing the application effect of the lipase. Because silkworms mainly eat mulberry leaves, oak leaves and the like to finish growth and development, how to improve the residual quantity of lipase on leaves and improve the rain wash resistance and the sunlight irradiation resistance is a key technology for improving the application effect of the lipase, and how to simply, conveniently and quickly apply the lipase is particularly important. As is known, the lipase powder is not easy to dissolve in water, the poor solubility in the using process causes the non-uniformity of spraying, most of enzyme liquid is dripped to the ground along leaves after spraying, the ductility and the permeability are poor, the enzyme liquid cannot be adhered to the leaves, the rain wash resistance is weak, the main component of the lipase is protein, and the enzyme activity is easy to lose after the sunlight irradiation.
Aiming at the problems, the invention provides a lipase wettable powder by referring to the processing mode of pesticide preparations.
The lipase wettable powder disclosed by the invention comprises the following raw materials in percentage by mass: 75-85% of lipase raw powder, 5-12% of wetting agent, 5-12% of dispersing agent, 1-3% of stabilizing agent and 1-3% of protective agent.
The wetting agent is selected from alkyl polyglycoside, span, tween or a mixture thereof.
The dispersant is calcium lignosulphonate or sodium lignosulphonate or white carbon black or a mixture thereof.
The stabilizer is Na2CO3Or K2HPO4Or carboxymethyl starch or xanthan gum or sodium citrate or a mixture thereof; the protective agent is ascorbic acidAcid or carboxymethyl cellulose or dextrin or a mixture thereof.
The lipase raw powder is derived from lipase in animals or plants or microorganisms, and is preferably derived from alkaline lipase produced by wild microorganisms or genetically engineered microorganisms. Lipase powders are commercially available and those skilled in the art can prepare lipase raw powders according to the methods disclosed in the literature.
The lipase raw powder is powdery lipase obtained by spray drying and freeze drying.
The lipase wettable powder mainly plays a role of a wetting agent, and the wetting agent is a substance capable of reducing liquid-solid interfacial tension, increasing the contact of liquid to a solid surface or increasing the wetting and spreading of the liquid to the solid surface. The wetting agent mainly improves the wetting dispersibility of the agent in water, improves the permeability of the drug solution, and improves the spreadability on the treatment object. The sunlight irradiation resistance is mainly realized by adding a stabilizing agent and a protective agent. The comprehensive effect of each component improves the residual quantity of the lipase on the leaves and improves the capability of resisting rain wash and sunlight irradiation.
The form of preparing the lipase zymogen powder into the wettable powder can ensure that the lipase zymogen powder is simple, convenient and quick to apply, compared with simple powder, the wetting agent and the dispersing agent are added into the wettable powder, and the wettable powder can be wetted and dispersed by water to form suspension after being added into water, so that the spraying is ensured to be more uniform in the spraying application process, and meanwhile, the lipase zymogen powder has strong adhesiveness and less drift after being sprayed and can better play a role.
The lipase wettable powder is prepared by the following steps: uniformly mixing the lipase raw powder, the wetting agent, the dispersing agent, the stabilizing agent and the protective agent in proportion, sieving, removing large particles, crushing by using an airflow crusher, and uniformly mixing to obtain the lipase preparation.
The lipase can act on the leaf surface of oak leaves and the like as a biocatalyst, hydrolyzes the structure of the leaf surface which is difficult to be decomposed by silkworms, hydrolyzes fat in the leaves, improves the utilization rate of nutrient components in the leaves, further improves the yield of silkworm cocoons, improves the disease resistance of the silkworms and improves the pupa building rate.
The lipase wettable powder is applied to silkworm breeding, and the specific steps are as follows: diluting the lipase wettable powder by 50-200 times with water according to the actual using amount, stirring until the lipase wettable powder is completely dissolved, spraying the lipase wettable powder on field mulberry leaves or oak leaves, and picking and feeding silkworms after drying; or spraying the dissolved lipase wettable powder on a proper amount of leaves, spreading and airing the leaves, and feeding silkworms.
Wherein, the airing is natural drying. After the leaves are naturally dried, the leaves are fed with feed to feed silkworms, so that the situation that the silkworms feed too much water to affect the digestion and absorption of the leaves can be avoided, and the feeding amount is determined according to the feeding amount of the silkworms.
The invention has the following advantages:
(1) the lipase provided by the invention is innovatively used as an additive for silkworm breeding, has obvious effects on improving the growth and development of silkworms, increasing the cocoon yield and the pupa health rate of the silkworms and other factors, is suitable for all bred silkworm varieties and all instars, and has universality.
(2) The lipase wettable powder disclosed by the invention is particularly suitable for spraying of leaves, can increase the adhesion of lipase on the leaves, and can prolong the lasting effect under natural conditions in the field.
(3) The preparation method of the lipase wettable powder has the characteristics of simple process, low cost, easy industrial production and the like, and the lipase preparation is safe, nontoxic and residue-free to the environment, people and livestock; simple and convenient use method, small application dosage, obvious yield increasing effect, easy popularization and application and the like.
Detailed Description
To further illustrate the invention, a series of examples are given below. These examples are purely illustrative and are intended to be a detailed description of the invention only and should not be taken as limiting the invention.
Example 1
Every 100g of lipase wettable powder contains 75.0g of lipase powder, 75.78 g of span-805.0 g, 805.0g of tween-805.0 g, 5.36g of white carbon black, 5.6g of sodium lignosulfonate, 1.04g of dipotassium hydrogen phosphate, 1.0g of sodium citrate and 2.0g of dextrin.
The lipase wettable powder is prepared by the following steps: mixing raw materials of lipase powder, span-80, tween-80, white carbon black, sodium lignosulfonate, dipotassium hydrogen phosphate, sodium citrate, dextrin and the like in proportion, pouring the mixture into a stirrer, stirring the mixture uniformly, sieving the mixture by using a 80-mesh sieve, removing large particles, crushing the mixture by using a 300-mesh airflow crusher, and mixing the crushed mixture uniformly.
Example 2
The lipase wettable powder contains 78.5g of lipase powder, 6.9g of alkyl polyglycoside (APG-1214), 5g of sodium lignosulfonate, 4g of white carbon black, 1.6g of sodium carbonate, 1g of sodium citrate and 3g of carboxymethyl cellulose in every 100g of the lipase wettable powder.
The lipase wettable powder is prepared by the following steps: mixing raw materials of lipase powder, alkyl polyglycoside (APG-1214), white carbon black, sodium lignosulphonate, sodium carbonate, sodium citrate, carboxymethyl cellulose and the like in proportion, pouring the mixture into a stirrer to be uniformly stirred, sieving the mixture by a 80-mesh sieve, removing large particles, crushing the mixture by a 300-mesh airflow crusher, and uniformly mixing the crushed mixture.
Example 3
100g of lipase wettable powder contains 80.0g of lipase powder, 5.04g of alkyl polyglycoside (APG-1214), 3.36g of white carbon black, 5.6g of calcium lignosulfonate, 2.0g of carboxymethyl starch, 1.0g of xanthan gum and 3.0g of ascorbic acid.
The lipase wettable powder is prepared by the following steps: raw materials of lipase powder, alkyl polyglycoside (APG-1214), white carbon black, calcium lignosulphonate, xanthan gum, carboxymethyl starch, ascorbic acid and the like are poured into a stirrer in proportion and stirred uniformly, then the mixture is sieved by a 80-mesh sieve to remove large particles, and the large particles are crushed by a 300-mesh airflow crusher and mixed uniformly.
Example 4
Every 100g of the lipase wettable powder contains 81.5g of lipase powder, 5.8g of alkyl polyglycoside (APG-1214), 3.4g of white carbon black, 4.8g of calcium lignosulfonate, 1.5g of sodium carboxymethylcellulose, 1.0g of sodium carbonate and 2.0g of carboxymethyl starch.
The lipase wettable powder is prepared by the following steps: mixing raw materials of lipase powder, alkyl polyglycoside (APG-1214), white carbon black, calcium lignosulphonate, sodium carboxymethylcellulose, sodium carbonate, carboxymethyl starch and the like in proportion, pouring the mixture into a stirrer, stirring the mixture uniformly, sieving the mixture by a 80-mesh sieve, removing large particles, crushing the mixture by a 300-mesh airflow crusher, and mixing the crushed mixture uniformly.
Example 5
Every 100g of the lipase wettable powder contains 85g of lipase raw powder, 805.5g of span-805.5 g of calcium lignosulfonate, 1.5g of carboxymethyl starch and 1.7g of sodium carboxymethyl cellulose.
The lipase wettable powder is prepared by the following steps: mixing raw materials of lipase powder, span-80, calcium lignosulfonate, carboxymethyl starch, sodium carboxymethyl cellulose and the like in proportion, pouring the mixture into a stirrer to be uniformly stirred, sieving the mixture by using a 80-mesh sieve, removing large particles, crushing the mixture by using a 300-mesh airflow crusher, and uniformly mixing the crushed mixture.
Experimental example 1 application of lipase preparation to silkworm breeding from 3 years old
The test sample is 100g of the lipase preparation prepared in example 1, and is diluted 50-fold, 100-fold and 200-fold with 5kg, 10kg and 20kg of water respectively to form 3 series concentrations. Collecting healthy folium Mori from mulberry, soaking in the above lipase solution with different concentrations for 5-10s, taking out, and air drying. Feeding the treated mulberry leaves to the 3-year-old silkworm until cocooning; the silkworm is fed with the aired mulberry leaves after being soaked in clear water as a blank Control (CK). And collecting 5-7 days after silkworm cocoons are formed, and respectively weighing and calculating indexes such as single-grain full cocoon weight, single-grain cocoon layer weight, pupa building rate and the like.
From the experimental results in table 1, feeding of the lipase preparation from 3 years old has a significant effect on the growth and development of the silkworm, and compared with a control, the indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa strengthening rate and the like are greatly improved, which shows that the lipase preparation has an obvious effect on increasing the yield of the full cocoons, and the yield is about 8%.
Experimental example 2 application of lipase preparation in silkworm breeding from 4 th instar
The test sample is 100g of the lipase preparation prepared in example 2, and is diluted into 3 series concentrations by adding 5kg of water, 10kg of water and 20kg of water respectively. Collecting healthy folium Mori from mulberry, soaking in the above lipase solution with different concentrations for 5-10s, taking out, and air drying. Feeding the treated mulberry leaves to 4-year-old silkworm respectively until cocooning; the silkworm is fed with the aired mulberry leaves after being soaked in clear water as a blank Control (CK). Collecting 5-7 days after silkworm cocoons are formed, and respectively weighing and calculating indexes such as whole cocoon weight, cocoon layer weight, pupa body weight, pupa strengthening rate and the like.
From the experimental results in table 2, feeding of the lipase preparation from 4 years old has an obvious effect on the growth and development of the silkworm, and compared with the control, the indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa strengthening rate and the like are greatly improved, which shows that the lipase preparation has an obvious effect on increasing the yield of the full cocoons, and the yield is about 7%.
Experimental example 3 application of lipase preparation to silkworm breeding at 5 th instar
The test sample is 100g of the lipase preparation prepared in example 3, and is diluted into 3 series concentrations by adding 5kg of water, 10kg of water and 20kg of water respectively. Collecting healthy folium Mori from mulberry, soaking in the above lipase solution with different concentrations for 5-10s, taking out, and air drying. Feeding the treated mulberry leaves to 5-year-old silkworm respectively until cocooning; the silkworm is fed with the aired mulberry leaves after being soaked in clear water as a blank Control (CK). Collecting 5-7 days after silkworm cocoons are formed, and respectively weighing and calculating indexes such as whole cocoon weight, cocoon layer weight, pupa body weight, pupa strengthening rate and the like.
From the experimental results in table 3, feeding of the lipase preparation from 5 years old has an obvious effect on the growth and development of the silkworm, and compared with the control, the indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa strengthening rate and the like are greatly improved, which shows that the lipase preparation has an obvious effect on increasing the yield of the full cocoons, and the yield is about 5%.
Experimental example 4 application of lipase preparation in tussah culture from 4 th instar
The experimental sample is 100g of the lipase preparation prepared in example 4, and 10kg of water is added, and the mixture is uniformly stirred to prepare a suspension. Spraying on oak tree, and feeding the processed oak tree leaves by 4-year-old tussah until cocoons are formed; feeding tussah as blank Control (CK) with the tussah treated with spray water. Collecting the cocoons of the tussah 5-7 days later, and respectively weighing and calculating indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa building rate and the like.
From the experimental results in table 4, the lipase preparation fed from 4 years old has significant influence on the growth and development of the tussah, and compared with the control, the indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa strengthening rate and the like are greatly improved, which shows that the lipase preparation has obvious effect on increasing the yield of the full cocoons, and the yield is about 14%.
Experimental example 5 application of lipase preparation in tussah cultivation from age 5
The experimental sample is 100g of the lipase preparation prepared in example 5, and 10kg of water is added, and the mixture is uniformly stirred to prepare a suspension. Spraying on oak tree, and feeding the treated oak tree leaves by 5-year-old tussah until cocoons are formed; feeding tussah as blank Control (CK) with the tussah treated with spray water. Collecting the cocoons of the tussah 5-7 days later, and respectively weighing and calculating indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa building rate and the like.
From the experimental results in table 5, the feeding of the lipase preparation from 5 years old has a significant effect on the growth and development of the tussah, and compared with the control, the indexes such as the weight of single-grain full cocoons, the weight of single-grain cocoon layers, the pupa strengthening rate and the like are greatly improved, which shows that the lipase preparation has an obvious effect on increasing the yield of the full cocoons, and the yield is increased by about 11%.
TABLE 1 influence of lipase preparations on the Whole cocoon weight, cocoon layer weight and pupa health rate of 3-instar silkworms
TABLE 2 influence of lipase preparations on the Whole cocoon weight, cocoon layer weight and pupa health rate of 4-instar silkworms
TABLE 3 influence of lipase preparations on the Whole cocoon weight, cocoon layer weight and pupa health rate of 5-instar silkworms
TABLE 4 influence of lipase preparations on the Whole cocoon weight, cocoon layer weight and pupa strengthening rate of tussah silkworms from the age of 4
TABLE 5 influence of lipase preparations on the Whole cocoon weight, cocoon layer weight and pupa strengthening rate of tussah silkworms from the age of 5
Claims (3)
1. A lipase wettable powder for silkworm breeding is characterized in that: the material composition is as follows by mass percent: 75-85% of lipase raw powder, 5-12% of wetting agent, 5-12% of dispersing agent, 1-3% of stabilizing agent and 1-3% of protective agent;
the wetting agent is selected from alkyl polyglycoside, span, tween or a mixture thereof;
the dispersant is calcium lignosulphonate or sodium lignosulphonate or white carbon black or a mixture thereof;
the lipase raw powder is derived from alkaline lipase produced by wild microorganisms or genetically engineered microorganisms;
the stabilizer is Na2CO3Or K2HPO4Or carboxymethyl starch or xanthan gum or sodium citrate or a mixture thereof; the protective agent is carboxymethyl cellulose or dextrin or a mixture thereof.
2. The lipase wettable powder for silkworm breeding according to claim 1, which is characterized in that: the lipase raw powder is powdery lipase obtained by spray drying and freeze drying.
3. The use of the lipase wettable powder for silkworm breeding of claim 1 in silkworm breeding, which is characterized in that: the method comprises the following specific steps: diluting the lipase wettable powder by 50-200 times with water according to the actual using amount, stirring until the lipase wettable powder is completely dissolved, spraying the lipase wettable powder on field mulberry leaves or oak leaves, and picking and feeding silkworms after drying; or spraying the dissolved lipase wettable powder on a proper amount of leaves, spreading and airing the leaves, and feeding silkworms;
wherein, the airing is natural drying.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57112333A (en) * | 1980-12-30 | 1982-07-13 | Katsuaki Tanaka | Medicine for promoting nutrition of silkworm |
| CN102575238A (en) * | 2009-12-01 | 2012-07-11 | 日清奥利友集团株式会社 | Lipase powder preparation and use thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57112333A (en) * | 1980-12-30 | 1982-07-13 | Katsuaki Tanaka | Medicine for promoting nutrition of silkworm |
| CN102575238A (en) * | 2009-12-01 | 2012-07-11 | 日清奥利友集团株式会社 | Lipase powder preparation and use thereof |
Non-Patent Citations (1)
| Title |
|---|
| 脂肪酶在禽畜养殖业中的应用于研究;杨滔等;《北方牧业》;20110605;第3结第2段 * |
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