Method for screening freeze-drying protective agent for lactobacilli by using liposome technology
A technology for drying protective agents and liposomes, which is applied in biochemical equipment and methods, and the determination/inspection of microorganisms, which can solve the problems of long time and high operation requirements.
Inactive Publication Date: 2014-03-26
BEIJING FORESTRY UNIVERSITY
View PDF1 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
The method of screening protective agents is mainly to use the traditional cell culture method to compare the changes in the number of viable bacteria, enzyme activity and other indicators before and after freeze-drying, which takes a long time and requires high operation
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0026] (1) Refer to step 1 to prepare β-galactosidase liposomes.
[0027] (2) Add different protective agents
[0028] Divide 60ml of enzyme liposome into five parts, and put them into test tubes, 10ml in each tube, labeled ①②③④⑤.
[0029] No. ① test tube, put it in the refrigerator to refrigerate;
[0030] In test tube ②, add trehalose. Prepare 100mg / 100ml. That is, 0.1000g, dissolved in 100ml of water. Then take 2.5ml to 10ml liposome solution. (The final trehalose concentration is about 20mg / 100ml)
[0031] No. ③ test tube, add hyaluronic acid. You can prepare 2mg / 100ml hyaluronic acid solution (weigh 0.0020g hyaluronic acid dissolved in 100ml water), and then take 2.5ml into 10ml liposome solution. (The final hyaluronic acid concentration is 0.4mg / 100ml)
[0032] In test tube ④, add trehalose and hyaluronic acid mutual protection agent, 10mg / 100ml and 0.2mg / 100ml. Take 1.286ml of 100mg / 100ml trehalose solution and 1.429ml of 2mg / 100ml hyaluronic acid solution.
...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
| Property | Measurement | Unit |
|---|---|---|
| particle size | aaaaa | aaaaa |
| dispersity | aaaaa | aaaaa |
Login to View More
Abstract
The invention discloses a method for simply and rapidly screening a freeze-drying protective agent for lactobacilli, which comprises the following steps of: firstly, preparing a beta-galactosidase liposome: dissolving soya bean lecithin and octadecylamine which are in molar ratio of 10:1 and taken as preparation materials into 15ml of diethyl ether, wherein the total amount of the soya bean lecithin and the octadecylamine is 144mumol; encapsulating 5ml of beta-galactosidase suspension, wherein the protein concentration of the beta-galactosidase suspension is 1.286mg / ml, and the enzymatic specific activity of the beta-galactosidase suspension is 1970U / mg of protein; the beta-galactosidase liposome is characterized in that the average grain diameter is 110nm to150nm, the polydispersity index (PDI) is 0.265 to 0.390, and the Zeta potential is -37.1mV to -42.0mV; then, adding different freeze-drying protective agents so as to perform freeze drying on the prepared liposome suspension, wherein the height of a liquid level is 0.5-1cm; prefreezing for 12 hours at the temperature which is lower than -25 DEG C; performing the freeze drying for 12 hours at the temperature of -50 to -40 DEG C at the vacuum degree of 40-80Pa; and finally, rehydrating the freeze-drying liposome, and measuring the change of the zymoprotein encapsulation rate. The protective agent with the minimum encapsulation rate reduction is best in effect. According to the protective agent screened by adopting the method disclosed by the invention, the survival rate of the lactobacilli in the freeze-drying process can be effectively increased.
Description
technical field [0001] The invention relates to liposome preparation and the field of microorganisms. Background technique [0002] Freeze-drying is to sublimate the ice of microbial cells to be preserved under vacuum conditions and frozen state, and finally achieve drying. Its steps are divided into 3 steps, the first is the pre-freezing process, the speed and temperature of pre-freezing are determined according to the type of microorganisms. The second is the sublimation process. The pre-frozen microorganisms are placed in a high vacuum environment, and the ice crystals absorb heat and directly turn into water vapor and overflow from the microorganisms. The sublimation process gradually progresses from the inside to the outside. After the ice crystal sublimates, many micropores are left in the microorganism body. The third is the analysis process, which resolves the bound water remaining in the microorganisms, further reducing the water content in the microorganisms. ...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/34C12Q1/02
Inventor 刘友群张柏林贾春凤展海宁
Owner BEIJING FORESTRY UNIVERSITY