CN109526873A - A kind of method of feeding caenorhabditis elegan grease or liposoluble substance - Google Patents
A kind of method of feeding caenorhabditis elegan grease or liposoluble substance Download PDFInfo
- Publication number
- CN109526873A CN109526873A CN201811222503.8A CN201811222503A CN109526873A CN 109526873 A CN109526873 A CN 109526873A CN 201811222503 A CN201811222503 A CN 201811222503A CN 109526873 A CN109526873 A CN 109526873A
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- CN
- China
- Prior art keywords
- grease
- liposoluble substance
- caenorhabditis elegan
- feeding
- elegan
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
Abstract
The present invention provides a kind of methods of feeding caenorhabditis elegan grease or liposoluble substance, it includes the following steps: to prepare certain density protein solution, grease or liposoluble substance are mixed with protein solution, it is prepared into lotion by means such as homogeneous, high speed shear or ultrasounds, finally above-mentioned emulsion is added in caenorhabditis elegan liquid culture solution according to a certain percentage and is cultivated.The present invention solves the problems, such as that grease or liposoluble substance can not efficiently be taken in by caenorhabditis elegan, and method is simple and efficient to handle, to provide solution using caenorhabditis elegan as the trophic function of model evaluation grease or liposoluble substance.
Description
Technology neighborhood
The invention belongs to biotechnology neighborhood, it is related to building for a kind of method of feeding caenorhabditis elegan grease or liposoluble substance
It is vertical.
Technical background
Caenorhabditis elegans (Caenorhabditis elegans, C.elegans) be it is a kind of it is nontoxic, can be independent
The nematode of existence.Its individual is small, and adult only 1.5mm long is hermaphroditic (hermaphrodites), male Jin Zhan group
0.2%, can self-fertilization or double sexual reproductions；The average history of life is 3.5 days at 20 DEG C, and average reproductive capacity is 300-350；
If but mate with male worm, it can produce up to 1400 or more offsprings.From nineteen sixty-five, scientist Sydney Brenner is utilized
Model organism of the nematode as molecular biology and Developmental Biology research field.
Edible oil is also referred to as " edible oil ", refers to and is making used in food products, animal or vegetable fat.Under room temperature
For liquid.Due to raw material sources, processing technology and quality etc., common edible oil is mostly vegetable fat, including maize
Oil, rapeseed oil, peanut oil, fiery sesame oil, corn oil, olive oil, camellia oil, palm oil, canola oil, sunflower oil, soybean oil,
Sesame oil, linseed oil (siritch), grape seed oil, walnut oil, peony seed oil etc..
Caenorhabditis elegan is as a kind of common model organism, due to its unique feeding habits (feed on bacteria) and life
Environment (hydrophilic culture medium), so that caenorhabditis elegan has many difficulties in terms of the nutrition of research grease.
Summary of the invention
To solve the above problems, the present invention provides a kind of methods of feeding caenorhabditis elegan grease or liposoluble substance.
The purpose of the present invention is what is be accomplished by the following way:
A kind of method of feeding caenorhabditis elegan grease or liposoluble substance, includes the following steps,
Step 1: protein solution is prepared；
Step 2: grease or liposoluble substance are added in step 1 in protein solution obtained, mixed solution is obtained；
Step 3: mixed solution obtained in step 2 is subjected to micronized, Homogenization Treatments, lotion is made；
Step 4: lotion made from step 3 is added in caenorhabditis elegan culture medium, carries out the culture of caenorhabditis elegan.
Preferably, the protein solution preparation method in step 1 are as follows: weigh a certain amount of albumen, sodium azide, addition is gone
Ionized water dissolution, stirring.
Preferably, the protein solution concentration prepared in step 1 is 0.01%~50%, the concentration of sodium azide is 0~
0.1%.
Preferably, the protein solution in step 1 is prepared by one or more albumen, and the albumen includes but is not limited to
Lactalbumin, casein sodium, casein, soybean protein, pea protein, ovalbumin, ovalbumin.
Preferably, the time that protein solution stirring is prepared in step 1 is 0.1~48 hour.
Preferably, the mass ratio of the grease in step 2 or liposoluble substance and protein solution is 1:100~50:100.
Preferably, the micronized in step 2, Homogenization Treatments include but is not limited to homogeneous, high speed shear or ultrasound.
Preferably, the rate of high speed shear is 5000~25000r/min, and the time of shearing is 0.5~30 minute.Ultrasound
Frequency be 20kHz~60kHz, intensity be 30-550W cm-2, the time is 0.1~10 minute, keeps low temperature.The pressure of homogeneous
For 20~300Mpa, homogeneous is primary or more, keep low temperature.
Preferably, the emulsion particle diameter range in step 3 is 100~500nm.
Preferably, in step 4 caenorhabditis elegan culture medium be caenorhabditis elegan liquid culture medium, lotion additional amount be 1%~
20%.Use albumen as lotion, the surface tension of water layer and oil reservoir can be reduced and reduces the surface of water layer and air
Power is easily formed stable lotion on the whole, can be added in nematode liquid culture medium, and lotion can be by caenorhabditis elegan
It eats.
The present invention solves grease or liposoluble substance and can not efficiently be taken in by caenorhabditis elegan in caenorhabditis elegan culture medium
The problem of, method is simple and quick, to be mentioned using caenorhabditis elegan as external model evaluation grease or liposoluble substance nutrition
For approach.
Detailed description of the invention
Fig. 1 is the influence of different ultrasonic times and ultrasonic power to emulsion particle diameter
Wherein, SCN is casein sodium；WPI is whey concn albumen；SPI is soybean protein.
Fig. 2 is the influence that various concentration oily in different-grain diameter and lotion takes in caenorhabditis elegan
Wherein, RFU/500worm is the fluorescent value of 500 caenorhabditis elegans；200nm, 300nm, 400nm are respectively lotion grain
Diameter is 200 nanometers, 300 nanometers, 400 nanometers.
The influence that the protein emulsion of the different types of oil preparation of Fig. 3 takes in caenorhabditis elegan
Wherein, RFU/500worm is the fluorescent value of 500 nematodes；MCT is sweet three ester of middle carbochain；Olive oil is olive
Oil；Soybean oil is soybean oil；Algal oil is algae oil.
Specific embodiment
Further technical solution of the present invention is illustrated below by way of case, it should be understood that be below only example of the present invention
Property explanation, do not limit to and the protection scopes of the claims in the present invention.
The basic skills of 1 feeding caenorhabditis elegan grease of embodiment
Casein sodium is dissolved in the distilled water of 0.02% sodium azide, 1 gram of casein sodium is dissolved in 100 milliliters of deionizations
In water, it is configured to protein solution, protein solution is stirred at room temperature 2 hours.
10g oil is weighed, 100 milliliters of protein solutions are added to.
By 20000 revs/min of mixed solution high speed shear, shear 2 minutes；Then, ultrasonic probe is goed deep into solution, the bottom of according to
Face 3mm, with the ultrasonic probe of 20kHz, intensity is 40~80W cm-2Under conditions of, carry out ultrasound.Handling the time is respectively 2
Minute, 3 minutes, 4 minutes, 5 minutes and 6 minutes, obtain lotion.
Embodiment 2: the experiment of protein emulsion property
Experiment and instrument: magnetic stirring apparatus, ultrasonic emulsification instrument, 2000 particles distribution instrument of Malvern, supercentrifuge are
Commercially available conventional instrument.
The measurement of protein emulsion average grain diameter
Protein emulsion prepared by Example 1, is measured by Malvern size distribution instrument.Made using deionized water
For dispersing agent, parameter is set as 25 DEG C of temperature, measures lotion emulsion droplet with Malvern ParticleSizer.
Fig. 1 be the influence of different ultrasonic times and ultrasonic power to emulsion particle diameter, it can be found that ultrasonic power difference and
The difference of ultrasonic time has a significant effect to the partial size of protein emulsion.
Embodiment 3: caenorhabditis elegan intake experiment
Experiment and instrument: magnetic stirring apparatus, ultrasonic emulsification instrument, 2000 particles distribution instrument of Malvern, fluorescence microplate reader, high speed
Centrifuge is commercially available conventional instrument.
Protein emulsion prepared by Example 1, is dyed by Nile red.Take 0.01Mm, 0.1Mm, 1mM dye cream
It is added in caenorhabditis elegan liquid culture medium, caenorhabditis elegan is added and feeds 24 hours.It is successfully taken in by the method caenorhabditis elegan
Grease.
Fig. 2 is the influence that various concentration oily in different-grain diameter and lotion takes in caenorhabditis elegan, it can be found that with cream
The intake of the increase caenorhabditis elegan of liquid concentration gradually increases.Intake with the increase caenorhabditis elegan of emulsion particle diameter gradually subtracts
It is small.
Fig. 3 is the influence that the protein emulsion of different types of oil preparation takes in caenorhabditis elegan, it can be found that caenorhabditis elegan
The intake of grease can be changed with the variation of oily type.
Claims (12)
1. a kind of method of feeding caenorhabditis elegan grease or liposoluble substance, which is characterized in that include the following steps,
Step 1: protein solution is prepared；
Step 2: grease or liposoluble substance are added in protein solution made from step 1, obtain mixed solution；
Step 3: mixed solution made from step 2 is subjected to micronized, Homogenization Treatments, lotion is made；
Step 4: lotion made from step 3 is added in caenorhabditis elegan culture medium, carries out the culture of caenorhabditis elegan.
2. the method for a kind of feeding caenorhabditis elegan grease according to claim 1 or liposoluble substance, which is characterized in that step
Protein solution preparation method in rapid one are as follows: weigh a certain amount of albumen, sodium azide, deionized water dissolving, stirring is added.
3. the method for a kind of feeding caenorhabditis elegan grease according to claim 1 or liposoluble substance, which is characterized in that step
The protein solution concentration prepared in rapid one is 0.01%~50%, and the concentration of sodium azide is 0~0.1%.
4. the method for a kind of feeding caenorhabditis elegan grease according to claim 1 or liposoluble substance, which is characterized in that step
The time that protein solution stirring is prepared in rapid one is 0.1~48 hour.
5. the method for a kind of feeding caenorhabditis elegan grease according to any one of claims 1-4 or liposoluble substance, special
Sign is that the protein solution in step 1 is prepared by one or more albumen, the albumen include but is not limited to lactalbumin,
Casein sodium, casein, soybean protein, pea protein, ovalbumin, ovalbumin.
6. the method for a kind of feeding caenorhabditis elegan grease according to claim 1 or liposoluble substance, which is characterized in that step
The mass ratio of grease or liposoluble substance and protein solution in rapid two is 1:100~50:100.
7. the method for a kind of feeding caenorhabditis elegan grease according to claim 1 or liposoluble substance, which is characterized in that step
Micronized, Homogenization Treatments in rapid two include but is not limited to homogeneous, high speed shear or ultrasound.
8. the method for a kind of feeding caenorhabditis elegan grease according to claim 7 or liposoluble substance, it is characterised in that high
The rate of speed shearing is 5000~25000r/min, and the time of shearing is 0.5~30 minute.
9. a kind of method of feeding caenorhabditis elegan grease or liposoluble substance according to claim 7, which is characterized in that ultrasound
Frequency be 20kHz~60kHz, intensity be 30-550W cm-2, the time is 0.1~10 minute, keeps low temperature.
10. a kind of method of feeding caenorhabditis elegan grease or liposoluble substance according to claim 7, which is characterized in that
The pressure of matter is 20~300Mpa, and homogeneous is primary or more, keeps low temperature.
11. the method for a kind of feeding caenorhabditis elegan grease or liposoluble substance, feature exist according to claim 1
In the emulsion particle diameter range in step 3 is 100~500nm.
12. the method for a kind of feeding caenorhabditis elegan grease or liposoluble substance, feature exist according to claim 1
In caenorhabditis elegan culture medium is caenorhabditis elegan liquid culture medium in step 4, and lotion additional amount is 1%~20%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811222503.8A CN109526873A (en) | 2018-10-19 | 2018-10-19 | A kind of method of feeding caenorhabditis elegan grease or liposoluble substance |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811222503.8A CN109526873A (en) | 2018-10-19 | 2018-10-19 | A kind of method of feeding caenorhabditis elegan grease or liposoluble substance |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114698735A (en) * | 2022-04-13 | 2022-07-05 | 华南理工大学 | Caenorhabditis elegans particle food and preparation method thereof |
Citations (4)
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| CN101941675A (en) * | 2009-07-05 | 2011-01-12 | 王旭 | Nanoparticles and preparation method thereof |
| CN103763916A (en) * | 2011-02-11 | 2014-04-30 | 孟山都技术公司 | Pesticidal nucleic acids and proteins and uses thereof |
| KR20160069200A (en) * | 2014-12-08 | 2016-06-16 | (주) 피러스 | Nematodes pesticide using natural material and manufacturing method of the same |
| CN108486200A (en) * | 2018-03-30 | 2018-09-04 | 合肥工业大学 | Acylation modification casein polypeptide, nanoemulsions, preparation method and application |
-
2018
- 2018-10-19 CN CN201811222503.8A patent/CN109526873A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101941675A (en) * | 2009-07-05 | 2011-01-12 | 王旭 | Nanoparticles and preparation method thereof |
| CN103763916A (en) * | 2011-02-11 | 2014-04-30 | 孟山都技术公司 | Pesticidal nucleic acids and proteins and uses thereof |
| KR20160069200A (en) * | 2014-12-08 | 2016-06-16 | (주) 피러스 | Nematodes pesticide using natural material and manufacturing method of the same |
| CN108486200A (en) * | 2018-03-30 | 2018-09-04 | 合肥工业大学 | Acylation modification casein polypeptide, nanoemulsions, preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| DANIEL COLMENARES等: "Delivery of dietary triglycerides to Caenorhabditis elegans using lipid nanoparticles: Nanoemulsion-based delivery systems", 《FOOD CHEMISTRY》 * |
| 吴克刚等: "《食品微胶囊技术》", 31 July 2006, 北京：中国轻工业出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114698735A (en) * | 2022-04-13 | 2022-07-05 | 华南理工大学 | Caenorhabditis elegans particle food and preparation method thereof |
| CN114698735B (en) * | 2022-04-13 | 2023-09-26 | 华南理工大学 | Caenorhabditis elegans particle food and preparation method thereof |
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| CB03 | Change of inventor or designer information |
Inventor after: Chang Ming Inventor after: Yang Jinyue Inventor after: Liu Ruijie Inventor after: Wang Xingguo Inventor after: Jin Qingzhe Inventor after: Zhang Tao Inventor after: Guo Xin Inventor before: Chang Ming Inventor before: Yang Jinyue Inventor before: Liu Ruijie Inventor before: Jin Qingzhe Inventor before: Zhang Tao Inventor before: Guo Xin |
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Application publication date: 20190329 |
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