Method for extracting mucus of S. gigantea used as antibiotic substance

The technology of an antibacterial substance and an extraction method is applied in the field of extracting an antibacterial substance from the mucilage of the giant sphenoid coral, and achieves the effects of avoiding substance inactivation, simple and convenient extraction process and low extraction cost.

Inactive Publication Date: 2007-08-01
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far no one has studied the extraction solvent of S. gigantea

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Frozen slime samples (50 ml) from S. gigantea were removed from the refrigerator and thawed at room temperature.

[0019] Prepare about 18ml of extraction solvent in a glass flask placed on an ice bath, and the proportioning ratio is:

[0020] Ultrapure water 8.5ml

[0021] 0.5M EDTA tetrasodium salt 1.2ml

[0022] 200mM phenylmethylsulfonyl fluoride 0.3ml

[0023] 0.36mM Aprotinin 0.05ml

[0024] Glacial acetic acid 8ml

[0025] Add 50 ml of the thawed mucus sample to the extraction solvent.

[0026] Stir for 3 hours in an ice bath, centrifuge (17000 rpm, 30 min, 4° C.), collect the supernatant, filter and ultrafilter. Add polyethylene glycol 8000 and glycine to the ultrafiltration concentrate to make the concentrations 1.5% and 0.2%, respectively. Freeze in a -70°C refrigerator, and then freeze-dry to obtain the finished product.

[0027] Weigh the freeze-dried product, dissolve it with 20 mM sodium phosphate buffer (pH7.2), and add the solution dropwise on a s...

Embodiment 2

[0029] Frozen slime samples (100 ml) from S. gigantea were removed from the refrigerator and thawed at room temperature.

[0030] Prepare about 50ml of extraction solvent in a glass flask placed on an ice bath, and the proportion is:

[0031] Ultrapure water 37.4ml

[0032] 0.5M EDTA tetrasodium salt 0.4ml

[0033] 200mM phenylmethylsulfonyl fluoride 0.1ml

[0034] 0.36mM Aprotinin 0.06ml

[0035] Glacial acetic acid 12ml

[0036] Add 100 ml of the thawed mucus sample to the extraction solvent.

[0037] Stir for 5 hours under ice bath condition, centrifuge (15000rpm, 40min, 5°C), collect the supernatant, filter and ultrafilter. Add polyethylene glycol 8000 and glycine to the ultrafiltration concentrate to make the concentrations 1% and 0.1%, respectively. Freeze in a -70°C refrigerator, and then freeze-dry to obtain the finished product.

[0038] Weigh the freeze-dried product, dissolve it with 20 mM sodium phosphate buffer (pH 7.0), and add the solution dropwise on a s...

Embodiment 3

[0040] Frozen slime samples (50 ml) from S. gigantea were removed from the refrigerator and thawed at room temperature.

[0041] Prepare about 20ml of extraction solvent in a glass flask placed on an ice bath, and the proportion is:

[0042] Ultrapure water 13ml

[0043] 0.5M EDTA tetrasodium salt 0.8ml

[0044] 200mM phenylmethylsulfonyl fluoride 0.15ml

[0045] 0.36mM Aprotinin 0.03ml

[0046] Glacial acetic acid 6ml

[0047] Add 50 ml of the thawed mucus sample to the extraction solvent.

[0048] Stir for 4 hours in an ice bath, centrifuge (20000rpm, 25min, 4°C), collect the supernatant, filter and ultrafilter. Add polyethylene glycol 8000 and glycine to the ultrafiltration concentrate to make the concentrations respectively 1.5% and 0.15%. Freeze in a -70°C refrigerator, and then freeze-dry to obtain the finished product.

[0049] Weigh the freeze-dried product, dissolve it with 20 mM sodium phosphate buffer (pH7.2), and add the solution dropwise on a sterilized filte...

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Abstract

A process for extracting the antibacterial substance from the mucilage of S. gigantea includes such steps as thawing said mucilage, proportionally adding it to the extracting solvent prepared from water, Na4EDTA, phenylmethyl sulfuryl fluoride, aprotinin and acetic acid, stirring on ice bath to obtain liquid extract, centrifugal of supernatant, filtering, ultrafiltering, concentrating, and freeze drying.

Description

technical field [0001] The invention relates to a method for extracting antimicrobial substances from the mucus of giant ligant coral (S. gigantea) Background technique [0002] Coral reef ecosystems are among the most productive and diverse systems on Earth, however, in recent decades many scientists have invested in the disease resistance of stony corals as disease outbreaks have killed coral polyps and led to reef bleaching. researching. The mucus secreted by coral polyps covers the surface of the reefs formed by coral polyps. This layer of mucus is the first gateway for corals to resist pathogens. Therefore, it is very important to study the anti-bacteria effects of mucus. The compounds contained in the mucus not only act as the coral's own anti-infection substances, but the biological activity of many compounds can also provide antimicrobials for humans. [0003] Although the study of mucus in stony corals is of great significance, the method of mucus extraction has r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N61/00A01P1/00
Inventor 陈国华黄良民谭烨辉
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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