CN109874788A - A kind of protein nano Liposomal formulation and its preparing the application in biological nano plant growth regulator - Google Patents
A kind of protein nano Liposomal formulation and its preparing the application in biological nano plant growth regulator Download PDFInfo
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- CN109874788A CN109874788A CN201910163819.2A CN201910163819A CN109874788A CN 109874788 A CN109874788 A CN 109874788A CN 201910163819 A CN201910163819 A CN 201910163819A CN 109874788 A CN109874788 A CN 109874788A
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Abstract
The present invention provides a kind of protein nano Liposomal formulation and its preparing the application in biological nano plant growth regulator.The protein nano Liposomal formulation is made of soybean lecithin, cholesterol, chitosan oligosaccharide and super quick albumen.Said preparation is that the nano-lipid carrier of super quick albumen is constructed by new bio nanotechnology, then chitosan oligosaccharide is coated on to the outer layer of lipid carrier by electrostatical binding, make the protein nano Liposomal formulation that there is stable nanostructure, not only increase the stability of super quick albumen, the chitosan oligosaccharide of external sheath not only increases the stability of nano-carrier simultaneously, also with super quick protein exhibits synergistic effect, common induction plant generates the resistance of wide spectrum, enhance the self-defence ability of plant, inhibit the growth of various plants pathogenic microorganism, and its is nontoxic and can be microbial degradation, pollution will not be generated to environment.The present invention provides a kind of good novel nano plant growth regulator of therapeutic effect.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of protein nano Liposomal formulation and its receive preparing biology
Application in rice plant growth regulator.
Background technique
Nanometer biotechnology refers to the nanotechnology for studying biological phenomena, it is the knot of nanotechnology and biology
It closes, while being also one and being related to physics, chemistry, quantum, mechanics, materialogy, electronics, computer, biology, doctor
The comprehensive cross discipline of etc. various fields.It is forward position and the hot issue in International Biotechnology field, is led in medical and health
Domain has a wide range of applications and specific industrialization prospect, especially nano-medicament carrier, nano biological sensor and imaging skill
Art and intelligent miniature medical instrument etc. will play a significant role in terms of the diagnosis of disease, treatment and health care.It is international
Upper nanometer biotechnology has obtained certain progress in the research of field of medicaments.
Nano structured lipid carrier is that by solid lipid and spatially incompatible liquid fatty mixes at a certain temperature
The nanoparticle drug administration carrier being prepared.By forming the lipid backbone of some compartments containing nanometer, drug delivery amount is improved, is avoided
Drug is squeezed in storing process, it with higher Drug loading capacity and reduces storage encapsulating compared with conventional carriers system
The problem of drug is revealed.
Summary of the invention
The object of the present invention is to provide a kind of protein nano Liposomal formulation and its preparing biological nano plant growth
Application in regulator.The preparation is that the nano-lipid carrier of super quick albumen is constructed by bio-nanotechnology, is then led to
It crosses electrostatical binding the outer layer that chitosan oligosaccharide is coated on lipid carrier is made.The protein nano Liposomal formulation has stable nanometer
Structure, said preparation invariance under the conditions of hydrochloric acid.
For achieving the above object, the present invention is achieved by the following scheme:
The present invention provides a kind of protein nano Liposomal formulations, and the preparation includes following components: in parts by weight, soybean phosphorus
It is 5-15 parts of rouge, 1-5 parts of cholesterol, 1-10 parts of chitosan oligosaccharide, quick albumen 1-10 parts super；The preparation passes through following preparation method system
:
(1) soybean lecithin and cholesterol are distributed in ethyl alcohol respectively, then are mixed with to obtain blank using ethyl alcohol volatility process
Nano-lipid carrier；
(2) super quick albumen is added in blank nano-lipid carrier at low temperature, carries out ultrasound or high speed shear, then incubates
Change；
(3) chitosan oligosaccharide is added after hatching a period of time, continues to hatch, obtains protein nano Liposomal formulation.
Further: the mass volume ratio concentration that soybean lecithin and cholesterol are distributed in ethyl alcohol in the step (1) is
10%-40%。
Further: the temperature range of low temperature is 10-25 DEG C in the step (2).
Further: chitosan is added after hatching 2 ~ 12h in the step (3).
Further: step (3) incubation temperature is 10-25 DEG C.
The present invention also provides the proteolipid body preparations in being used to prepare biological nano plant growth regulator
Application.
Further: use mass concentration ratio of the proteolipid body preparation in the plant growth regulator is
0.1-10%。
Further: the proteolipid body preparation is directly dissolved in water, front or back of the Direct spraying on blade face；Or
Seed soaking, root dipping, root application.
Further: the plant includes shallot, tobacco leaf, green pepper, soybean, cotton and tealeaves.
Compared with prior art, the advantages and benefits of the present invention are: protein nano lipid system provided by the invention
The not simple physical combination of agent constructs the nano-lipid carrier of super quick albumen by new bio nanotechnology, so
Chitosan oligosaccharide is coated on to by electrostatical binding the outer layer of lipid carrier afterwards, there is the biological nano plant growth regulator and stablize
Nanostructure, on the one hand improve the stability of super quick albumen, improve its stability twice, on the other hand, external sheath
Chitosan oligosaccharide not only increase the stability of nano-carrier, while also inducing plant jointly with super quick protein exhibits synergistic effect
The resistance for generating wide spectrum, the self-defence ability for enhancing plant, the growth for inhibiting various plants pathogenic microorganism, and its is nontoxic
And can be microbial degradation, pollution will not be generated to environment.The present invention provides a kind of with high content of technology and therapeutic effect is good
Novel nano plant growth regulator.
Detailed description of the invention
Fig. 1 is the particle size determination result schematic diagram of the super quick protein solution of blank of non-filter membrane；
Fig. 2 is the potential measurement result schematic diagram of the super quick protein solution of blank of non-filter membrane；
Fig. 3 was the particle size determination result schematic diagram of the super quick protein solution of the blank of 0.8 μm of film；
Fig. 4 was the potential measurement result schematic diagram of the super quick protein solution of the blank of 0.8 μm of film；
Fig. 5 was the blank liposome particle size determination result schematic diagram of 0.8 μm of film；
Fig. 6 was the blank liposome potential measurement result schematic diagram of 0.8 μm of film；
Fig. 7 was the super quick proteolipid diameter of carrier measurement result schematic diagram of 0.8 μm of film；
Fig. 8 was the super quick proteolipid carrier potential measurement result schematic diagram of 0.8 μm of film；
Fig. 9 was the super quick 01 particle size determination result schematic diagram of protein nano lipid carrier of the chitosan oligosaccharide modification of 0.8 μm of film；
Figure 10 was the super quick 01 potential measurement result schematic diagram of protein nano lipid carrier of the chitosan oligosaccharide modification of 0.8 μm of film；
Figure 11 was the super quick 02 particle size determination result schematic diagram of protein nano lipid carrier of the chitosan oligosaccharide modification of 0.8 μm of film；
Figure 12 was the super quick 02 potential measurement result schematic diagram of protein nano lipid carrier of the chitosan oligosaccharide modification of 0.8 μm of film；
Figure 13 is proteoliposome preparation nature measurement result schematic diagram；
Figure 14 is proteolipid body preparation and deionized water or hydrochloric acid reaction result schematic diagram, wherein being successively from left to right egg
White original solution adds water, albumen original solution to add hydrochloric acid, proteolipid liquid solution (soybean lecithin: chitosan oligosaccharide=4:1) plus water, proteolipin
Plastid solution (soybean lecithin: chitosan oligosaccharide=4:1) plus hydrochloric acid, proteolipid liquid solution (soybean lecithin: chitosan oligosaccharide=2:1) add
Water, proteolipid liquid solution (soybean lecithin: chitosan oligosaccharide=2:1) plus hydrochloric acid.
Figure 15 is the experimental result comparison diagram that proteolipid body preparation is used for shallot, wherein right side is existing general proteins,
Left side is proteolipid body preparation of the present invention.
Figure 16 is the experimental result comparison diagram that proteolipid body preparation is used for tobacco leaf, wherein upside is existing general proteins,
Downside is proteolipid body preparation of the present invention.
Figure 17 is the experimental result comparison diagram that proteolipid body preparation is used for soybean, wherein left side is existing general proteins,
Right side is proteolipid body preparation of the present invention.
Figure 18 is the experimental result comparison diagram that proteolipid body preparation is used for green pepper, wherein right side is existing general proteins,
Left side is proteolipid body preparation of the present invention.
Specific embodiment
Below by the drawings and specific embodiments, the technical scheme of the present invention will be explained in further detail.
Embodiment 1
The present invention provides a kind of protein nano Liposomal formulations comprising following components: 5-15 parts of soybean lecithin, cholesterol
1-5 parts, it is 1-10 parts of chitosan oligosaccharide, quick albumen 1-10 parts super, in parts by weight.
The preparation method of the protein nano Liposomal formulation specifically includes the following steps:
(1) first by 1-5 parts of 5-15 parts of soybean lecithin, cholesterol addition ethyl alcohol, low-temperature heat volatilizes ethyl alcohol, is added 10-
100 times of water, oscillator shake 10-30 minutes, prepare blank liposomes carrier；
(2) the super quick albumen of above-mentioned dosage is added to the blank nano-lipid carrier by (25 degrees centigrade) at low temperature
In, ultrasound or high speed shear are carried out, is then hatched；
(3) appropriate chitosan oligosaccharide is added after hatching 2-12h, continues to hatch, obtains protein nano Liposomal formulation.And it measures not
With the nanometer particle size and current potential at time point.
Embodiment 2
1, the Stability Determination of the biological nano plant growth regulator albumen
(1) measurement of the partial size and current potential of the super quick protein solution of blank, wherein partial size when not crossing film is 2037.9nm, sees figure
1；Current potential is -15.60mv, sees Fig. 2.Partial size after crossing 0.8 μm of film is 1504.1nm, sees Fig. 3；Current potential is -12.56mv, is seen
Fig. 4.
(2) measurement of liposomal particle size and current potential, wherein blank liposome is the blank liposomes of the super quick albumen of unentrapped
Body, the partial size for crossing 0.8 μm of film is 273.8nm, sees that Fig. 5, current potential are -20.33mv, sees Fig. 6.Contain the lipid nanometer of super quick albumen
The partial size that carrier crosses 0.8 μm of film is 699.8nm, sees that Fig. 7, current potential are -3.85mv, sees Fig. 8.Lipid carries after proof contains albumen
Body partial size becomes larger, and significant change also has occurred in current potential.
(3) chitosan oligosaccharide (it is 5%-10% that chitosan oligosaccharide, which accounts for the mass ratio of the proteoliposome) the super quick protein nano lipid of modification
Carrier 01, the partial size for crossing 0.8 μm of film is 570.9nm, sees that Fig. 9, current potential are -6.50mv, sees Figure 10.
(4) chitosan oligosaccharide (it is 1%-3% that chitosan oligosaccharide, which accounts for the mass ratio of the proteoliposome) the super quick protein nano lipid of modification
Carrier 02, the partial size for crossing 0.8 μm of film is 456.8nm, sees that Figure 11, current potential are -7.25mv, sees Figure 12.
Provable through experimental result: super quick albumen is wrapped in inside lipid carrier, and the protein nano lipid system
The partial size of agent is able to maintain stabilization within nanometer range.
2, the stability preliminary assessment of the protein nano Liposomal formulation
(1) its stability is observed after placing blank liposome and the liposome for containing albumen being prepared five days, is found
There is not obvious sediment phenomenon in the Liposomal formulation for containing albumen, sees Figure 13, it is known from figures that preparation stabilization obtained,
Meet requirement.20 and 40 in Figure 13 represent 20 DEG C and 40 DEG C of temperature, illustrate at 40 DEG C, contain receiving for albumen
The stability of the grain of rice is apparently higher than general proteins solution group, this is significant for transport of summer temperature when excessively high and storage,
In this case the holding time that can significantly extend albumen, cost is reduced, better curative effect is played.
(2) method of dropwise addition hydrochloric acid is chosen to detect the advantage and stability of the protein nano Liposomal formulation
Concrete operations are as follows: take each two parts of the different solutions of equivalent to be put into six identical cillin bottles, be added equivalent go from
Sub- water or hydrochloric acid, shaking, observe and record phenomenon, as a result as shown in figure 14.It is successively from left to right super quick albumen original solution
Water, super quick albumen original solution is added to add hydrochloric acid, super quick proteolipid liquid solution (soybean lecithin: chitosan oligosaccharide=4:1) plus water, super quick egg
White liposome solutions (soybean lecithin: chitosan oligosaccharide=4:1) plus hydrochloric acid, super quick proteolipid liquid solution (soybean lecithin: chitosan oligosaccharide=
2:1) plus water, super quick proteolipid liquid solution (soybean lecithin: chitosan oligosaccharide=2:1) add hydrochloric acid.It can clearly be seen from Figure 14
It observes, albumen original solution is easy denaturation under the conditions of hydrochloric acid, and manufactured nanometer formulation solution does not allow mutability；Nanometer formulation is molten
Liquid then obviously has better stability than albumen original solution, and liposome has excellent protective effect to albumen.
Embodiment 3
The proteolipid body preparation is directly dissolved in clear water (pH is neutral, 7-8 or so), Direct spraying blade face front or
Reverse side；It can also soak seed, root dipping, root application, dilute 1000 times of uses with clear water, the specification of dosage (liquid) is 15mL/100mL/
The quality of 250mL/500mL(wherein super quick albumen is than content 0.1-10%).
Use age: it can be used from seed-soaking to water content in harvest.It can make with other acidic fertiliziers or pesticide cooperation
With.One season crop generally spray 3-5 times, spray twice midfeather 15-20 days.This product is please deposited in into shady and cool, ventilation, drying
Place.It is not used together with strong acid, highly basic, strong ionic preparation or solvent.This product is nontoxic, other special anti-without making
Shield.
Several plant, experimental result are applied to using the proteolipid body preparation as biological nano plant growth regulator
As shown in figures 15-18, it can be seen from the figure that proteolipid body preparation of the present invention is used for shallot, tobacco leaf, green pepper and big
When beans, hence it is evident that the plant growth for applying the proteolipid body preparation is more vigorous, and insect pest ability improves, and blade face is more sent out
It is green, eugonic situation is shown, autoimmunity is improved.
Suitable for the industrial crops such as field crop and vegetables, melon and fruit, cotton, tobacco, tealeaves, flowers, in kind of a phase, seedling
Phase, florescence and fruiting period using effect are fine, and germination percentage is high, root is more, height of seedling, robust seedling, blade is full, chlorophyll content is high.?
The upper use such as capsicum, tobacco, soybean is preferable, has its unique mechanism of action and apparent diseases prevention, pest-resistant, effect of increasing production.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than is limited；Although referring to aforementioned reality
Applying example, invention is explained in detail, for those of ordinary skill in the art, still can be to aforementioned implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features；And these are modified or replace
It changes, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (9)
1. a kind of protein nano Liposomal formulation, it is characterised in that: the preparation includes following components: in parts by weight, soybean
It is -15 parts of phosphatidase 5,1-5 parts of cholesterol, 1-10 parts of chitosan oligosaccharide, quick albumen 1-10 parts super；The preparation passes through following preparation method
It is made:
(1) soybean lecithin and cholesterol are distributed in ethyl alcohol respectively, then are mixed with to obtain blank using ethyl alcohol volatility process
Nano-lipid carrier；
(2) super quick albumen is added in blank nano-lipid carrier at low temperature, carries out ultrasound or high speed shear, then incubates
Change；
(3) chitosan oligosaccharide is added after hatching a period of time, continues to hatch, obtains protein nano Liposomal formulation.
2. protein nano Liposomal formulation according to claim 1, it is characterised in that: soybean lecithin in the step (1)
The mass volume ratio concentration being distributed in ethyl alcohol with cholesterol is 10%-40%.
3. protein nano Liposomal formulation according to claim 1, it is characterised in that: the temperature of low temperature in the step (2)
Spending range is 10-25 DEG C.
4. protein nano Liposomal formulation according to claim 1, it is characterised in that: after the step (3) hatches 2 ~ 12h
Chitosan is added.
5. protein nano Liposomal formulation according to claim 1, it is characterised in that: step (3) incubation temperature is
10-25℃。
6. the described in any item proteolipid body preparations of claim 1-5 are in being used to prepare biological nano plant growth regulator
Application.
7. proteolipid body preparation according to claim 6 is being used to prepare answering in biological nano plant growth regulator
With, it is characterised in that: use mass concentration ratio of the proteolipid body preparation in the plant growth regulator is 0.1-
10%。
8. proteolipid body preparation according to claim 6 is being used to prepare answering in biological nano plant growth regulator
With, it is characterised in that: the proteolipid body preparation is directly dissolved in water, front or back of the Direct spraying on blade face；Or leaching
Kind, root dipping, root application.
9. proteolipid body preparation according to claim 6 is being used to prepare answering in biological nano plant growth regulator
With, it is characterised in that: the plant includes shallot, tobacco leaf, green pepper, soybean, cotton and tealeaves.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111758744A (en) * | 2020-06-23 | 2020-10-13 | 江苏省农业科学院 | Application of hypersensitive protein in improving content of theanine and amino acid in tea leaves |
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