Induction method of lysozyme amyloid fiber model

A fiber model and amyloid technology, applied in biochemical equipment and methods, enzymes, hydrolytic enzymes, etc., can solve the problem of non-toxicity of amyloid fibers, and achieve the effect of prolonging experimental operability, saving costs, and prolonging operation time

Pending Publication Date: 2020-03-24
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AI-Extracted Technical Summary

Problems solved by technology

However, egg white lysozyme is a normal protein, and the amyloid fibers formed are not t...
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The invention discloses an induction method of a lysozyme amyloid fiber model. The induction method comprises the following steps: (1) selecting lysozyme; (2) preparing an induction buffer solution; (3) preparing a lysozyme solution with a certain concentration; and (4) inducing at different temperatures. According to the method, high-activity lysozyme is dissolved in a glycine-hydrochloric acid buffer system, strong acid conditions are artificially created, and conversion of a lysozyme protein secondary structure is facilitated; during high-temperature induction, protein can be mistakenly folded to form amyloid fiber aggregates, the amyloid fiber model can be better built, the success rate of lysozyme amyloid fibrillation can be increased, and the obtained fibers are higher in toughness.Experiment errors caused by environmental problems can be reduced, no batch difference is caused by the induction method, repeatability and stability of data obtained through the model are improved, and the method is simple and beneficial to popularization.

Application Domain


Technology Topic

Protein proteinBiophysics +11


  • Experimental program(1)

Example Embodiment

[0018] The following specific embodiments will further clarify the present invention. It should be understood that the following specific embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. This patent provides a variety of parallel schemes, and the different expressions belong to an improved scheme based on the basic scheme or a parallel scheme. Each program has its own unique characteristics.
[0019] A method for inducing a lysozyme amyloid fiber model includes the following steps:
[0020] (1) Choose lysozyme: choose egg white lysozyme as the material for amyloid fibrosis model;
[0021] (2) Preparation of induction buffer: Weigh a certain amount of glycine solid particles, dissolve it in water, adjust the pH with HCl, and prepare it into a glycine-hydrochloric acid mother liquor for use;
[0022] (3) Preparation of a certain concentration of lysozyme solution: Weigh a certain amount of egg white lysozyme powder and dissolve it in proportion;
[0023] (4) Different temperature induction: After shaking and inducing for a certain period of time under high temperature conditions, it is transferred to low temperature conditions and allowed to stand for a certain period of time;
[0024] In step (1), the egg white lysozyme is directly purchased, but not limited to purchase, and laboratory-purified egg white lysozyme can also be used.
[0025] In step (2), the glycine is a molecular grade reagent, but it is not limited to this. Analytical grade glycine can be used and filtered after preparation. The filter membrane has a pore size of 0.22-0.45 μm; a certain concentration is 40-60 mM.
[0026] In step (2), the water is ultrapure water or sterile water, which can be self-made or purchased; both concentrated hydrochloric acid and dilute hydrochloric acid can be used for hydrochloric acid, and the pH is adjusted to 1.8-12.
[0027] In step (3), the certain concentration of egg white lysozyme is 20-200 μM.
[0028] In step (4), the high temperature condition is 55-68°C, the induction time is 0.5-24h, and the shaking speed is 180-300rpm. Different temperatures can be selected for high temperature conditions.
[0029] In step (4), the low temperature condition is 2-6°C, and the induction time is 6-20 days.
[0030] A more specific implementation is as follows: Example:
[0031] (1) Choose lysozyme: use egg white lysozyme powder purchased by MCE Biological Company.
[0032] (2) Select the induction buffer: select molecular grade glycine from MP Biotech, weigh 0.375g, and completely dissolve it in 40mL homemade ultrapure water, adjust the pH to 2.0 with dilute hydrochloric acid, and set the volume to 100mL, and configure it as a 50mM buffer. use.
[0033] (3) Preparation of a certain concentration of lysozyme solution: Weigh 0.0143 g of egg white lysozyme powder purchased from MCE Company, and dissolve it completely in 10 mL of glycine-hydrochloric acid buffer to prepare a 100 μM sample.
[0034] (4) Different temperature induction: induction at 65°C and 250 rpm for 8 hours, transfer to 4°C refrigerator for 8 days, observation under a transmission electron microscope to obtain amyloid fibers, indicating successful induction.


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