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A method for extracting bacteriostatic composition from waste materials produced by LAL

A technology for producing waste and limulus reagents, which is applied in botany equipment and methods, chemicals for biological control, fungicides, etc., can solve the problems of high cost, low limulus peptide content, cumbersome steps, etc., and achieve the purpose of inhibiting bacteria and the growth of fungi, avoid drug resistance and allergies, and have no toxic and side effects

Active Publication Date: 2020-05-05
GENOBIO PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] If the limulus peptide is directly extracted from the limulus cells, compared with the limulus peptide extracted from the limulus reagent waste, the finally obtained limulus peptide content is higher, but due to the conflict with the raw material (lysus cells) in the extraction of the above-mentioned limulus reagent, makes the extraction itself expensive
If the limulus peptide is directly extracted from the waste of the limulus reagent, the content of the limulus peptide is relatively low, and the subsequent purification steps are cumbersome and the process cost is high. is a big difficulty

Method used

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  • A method for extracting bacteriostatic composition from waste materials produced by LAL
  • A method for extracting bacteriostatic composition from waste materials produced by LAL

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Effect test

preparation example Construction

[0043] (2) Preparation of Limulusin Peptide Crude Extract

[0044] a. Acid hydrolysis: Add the lower layer sediment after centrifugation of the Limulus cell emulsion to the acid solution (20mmol / L) at a solid-liquid ratio of 1:6.5-8, and ultrasonically crush it at low temperature. The crushing conditions are: the ultrasonic output power per 100ml is 700W , The total ultrasonic action time is 10min, and each ultrasonic action time is 10S. Repeat the ultrasound 3 times, and collect the supernatant as the cell acid extraction solution;

[0045] b. Adjust the pH to remove precipitation: adjust the pH of the extract to 6.5 with NaOH, centrifuge to remove the precipitate, and obtain the supernatant;

[0046] c. Heat denaturation treatment: treat the supernatant in a boiling water bath for 5-10 minutes, centrifuge the precipitate at 4°C to collect the supernatant, and filter the supernatant with a 0.22uM membrane to obtain a crude lisectin peptide extract;

[0047] (3) Preparation ...

example 1

[0055] (1) Raw material handling

[0056] a. Collect Limulus whole blood,

[0057] b. Separation of Limulus blood cells and plasma, wherein the Limulus blood cells are first used in the production of Limulus reagent, and in the process of lysing and separating the Limulus blood cells in the Limulus reagent production process, the lower layer of the discarded sediment after the emulsion is centrifuged is used as a raw material for subsequent use. Plasma discarded during the production of Limulus reagent is used as raw material for backup;

[0058] (2) Preparation of Limulusin Peptide Crude Extract

[0059] a. Acid hydrolysis: add the lower precipitate of the centrifuged Limulus cell emulsion to the acid solution (20mmol / L) at a solid-liquid ratio of 1:6.5, and ultrasonically crush it at low temperature. The crushing conditions are: ultrasonic output power of 700W per 100ml, ultrasonic The total action time is 10min, and each ultrasonic action time is 10S. Repeat the ultrasou...

example 2

[0069] (1) Raw material handling

[0070] a. Collect Limulus whole blood,

[0071] b. Separation of Limulus blood cells and plasma, wherein the Limulus blood cells are first used in the production of Limulus reagent, and in the process of lysing and separating the Limulus blood cells in the Limulus reagent production process, the lower layer of the discarded sediment after the emulsion is centrifuged is used as a raw material for subsequent use. Plasma discarded during the production of Limulus reagent is used as raw material for backup;

[0072] (2) Preparation of Limulusin Peptide Crude Extract

[0073] a. Acid hydrolysis: add the lower precipitate of the Limulus cell emulsion to the acid solution (20mmol / L) at a solid-liquid ratio of 1:8, and ultrasonically crush it at low temperature. The total action time is 10min, and each ultrasonic action time is 10S. Repeat the ultrasound 3 times, and collect the supernatant as the cell acid extraction solution;

[0074] b. Adjust...

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Abstract

The invention relates to a method used for extracting an antibacterial composition from limulus reagent production waste materials. The method comprises following steps: 1, raw material treatment is carried out, wherein a lower layer precipitate waste material obtained in centrifugation of an emulsified product in limulus reagent production is collected, and is taken as a raw material I, and discarded blood plasma in limulus reagent production is collected, and is taken as a raw material II; 2, extraction of a tachyplesin crude extracted solution is carried out, wherein acidolysis is carried out, pH is adjusted so as to remove a precipitate product, and thermal denaturation is carried out; 3, preparation of a hemocyanin crude extracted solution from blood plasma is carried out; 4, preparation of a SOD crude extracted solution is carried out; 5, 60 to 80 parts of the tachyplesin crude extracted solution,10 to 20 parts of the hemocyanin crude extracted solution, and 10 to 20 parts of the SOD crude extracted solution are mixed, pH value is adjusted to 4 to 5, and filtering is carried out so as to obtain the antibacterial composition. According to the method, the crude extracted solutions are prepared rapidly and conveniently at low cost; the antibacterial composition is capable of inhibiting growth of a plurality of kinds of bacteria and fungus; compared with conventional chemical antibacterial products, the antibacterial composition possesses following advantages: the antibacterial composition is safe, mild, and effective, no toxic or side effect is caused, and generation of drug resistance and drug allergy caused by conventional antibacterial products can be avoided.

Description

technical field [0001] The invention relates to the technical field of re-extraction of Limulus reagent waste, in particular to a method for extracting an antibacterial composition from Limulus reagent production waste. Background technique [0002] Limulus, also known as horseshoe crab, is a large benthic arthropod living in the ocean. It has a history of more than 400 million years since the Ordovician in the early Paleozoic Era. It is a "living fossil" with a unique evolutionary status in the animal kingdom. one. Limulus is an excellent material for invertebrate hematology research, and it is also the main raw material for the development and utilization of Limulus resources and the development of Limulus reagents. It has been highly valued by scholars in Japan, the United States and other countries. With the study of the immune function of Limulus hemolymph and the immunological comparison with other invertebrate hemolymph, especially many antibacterial and antiviral ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N63/10A01N37/46A01P1/00A01P3/00
CPCA01N63/10
Inventor 辛鹤林张文倩曹佳利韩钊
Owner GENOBIO PHARM CO LTD