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Cell lysis self-assembly polypeptide compound, self-assembly method, self-assembly polypeptide preparation and application

A polypeptide complex, self-assembly technology, applied in the biological field, to achieve strong antibacterial and anti-inflammatory effects, improve assembly efficiency, and reduce critical concentration

Active Publication Date: 2022-06-07
HANGZHOU UMOTOR BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these non-covalent bonds are weak, they can still effectively drive self-assembly by relying on the advantages of the number of polypeptide molecules

Method used

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  • Cell lysis self-assembly polypeptide compound, self-assembly method, self-assembly polypeptide preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The self-assembly method of cell lysing polypeptide complex comprises the following steps:

[0043] S1, cell lysis

[0044] Get the fermentation broth at 4,000rpm, 30min, under 4 ℃ of conditions, centrifuge to collect thalline; use homogenate buffer (20mmol / LTris-Cl pH8.0, 100mmol / L NaCl, 2mmol / L MgCl 2 , 1 mmol / L DTT) to resuspend the bacteria, centrifuge at 6,000 rpm, 10 min, and 4 °C to remove the supernatant; add homogenization buffer to resuspend the pellet (1 g of pellet is added to 10 ml of homogenization buffer), and then add the final concentration of 1 mmol / L PMSF, sonicated in ice bath for 30 min (amplitude 55%, sonicated for 9 s, stopped for 5 s) to obtain cell lysate.

[0045] S2, centrifugal

[0046] The cell lysate was centrifuged at 4,000 rpm, 30 min, and 4°C to obtain a supernatant.

[0047] S3, enzymatic hydrolysis

[0048] Add 0.5% wt protease to the supernatant, and carry out enzymolysis at 55°C to obtain a supernatant enzymolysis solution; speci...

Embodiment 2-11

[0052] The difference from Example 1 is that the raw materials and dosage of the fermentation substrate are different, as shown in Table 1 for details.

Embodiment 1-11

[0053]Example 1-11 Raw materials and dosage table of fermentation substrates

[0054] Calcium chloride / mM Manganese sulfate / mM Copper sulfate / mM Arginine / mM Lysine / mM Glutamate / mM Example 1 0.1 0.1 0.1 / / 0.01 Example 2 10 0.1 0.1 / / / Example 3 50 0.1 0.1 / / / Example 4 10 50 0.1 / / / Example 5 10 100 0.1 / / 0.1 Example 6 10 50 30 / / / Example 7 10 50 60 / / / Example 8 10 50 / 0.01 / 0.2 Example 9 10 50 / / 0.01 / Example 10 10 50 / / 50 10 Example 11 10 50 / / 100 /

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Abstract

The invention relates to a cell lysis self-assembly polypeptide compound, a self-assembly method, a self-assembly polypeptide preparation and application, and relates to the field of biotechnology. The invention discloses a cell lysis polypeptide compound self-assembly method, which comprises the following steps: carrying out cell lysis, centrifugation and protease enzymolysis on thermophilic thermophilic bacteria to obtain supernatant enzymatic hydrolysate, adding the supernatant enzymatic hydrolysate into thermophilic thermophilic bacteria or thermophilic thermophilic bacteria fermentation liquor, and carrying out fermentation self-assembly to obtain the cell lysis self-assembly polypeptide compound. The method has the effects of improving the polypeptide self-assembly efficiency, improving the activity of cell lysis polypeptide and resisting bacteria and diminishing inflammation.

Description

technical field [0001] The present application relates to the field of biotechnology, in particular to a cell lysis self-assembled polypeptide complex, a self-assembly method, a self-assembled polypeptide preparation and application. Background technique [0002] The self-assembly of polypeptide molecules is a process in which the molecules constituting the system spontaneously assemble, arrange and integrate through a series of non-covalent forces under the condition of thermodynamic equilibrium, and finally form a uniform and ordered self-assembly. The self-assembly of polypeptides mainly depends on the mutual restriction of various non-covalent forces within the molecule and the result after reaching equilibrium. These interactions include hydrophobic interactions, hydrogen bonds, electrostatic interactions, and element-element conjugation. Although these non-covalent bond forces are weak, they can still effectively drive self-assembly by virtue of the advantage of the nu...

Claims

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Application Information

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IPC IPC(8): C12P21/06C12N1/20A61K8/64A61P29/00A61P37/08A61Q19/00A61Q19/08A23L33/18C12R1/01
CPCC12P21/06C12N1/20A61K8/64A61Q19/00A61Q19/005A61Q19/08A61P29/00A61P37/08A23L33/18A61K2800/85A23V2002/00A23V2200/30A23V2200/304A23V2200/302A23V2200/318A23V2250/55
Inventor 颜贵卉娄兰兰奕志英张明洲吴雪昌王旻子姚雨辰魏建良
Owner HANGZHOU UMOTOR BIOTECH CO LTD
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