Method for extracting extracellular protein of biological membrane by using quartz sand in cooperation with nonionic surfactant

Abstract

The invention provides a method for extracting extracellular protein of a biological membrane by using quartz sand in cooperation with a nonionic surfactant. The method comprises the following steps:(1), acquiring an anaerobic ammonium oxidation bacteria biological membrane in situ, performing homogenizing, performing washing and filtering to obtain wet mud; (2), mixing the wet mud with quartz sand and a nonionic surfactant with the mass percent concentration of 0.01%-0.1% at a temperature of 0-4 DEG C, and performing stirring at a low speed to obtain a mud-sand mixed solution; (3), carryingout high-speed centrifugal treatment on the mud-sand mixed solution at a temperature of 0-4 DEG C, taking a supernatant, performing filtering and discarding dregs; and (4), carrying out medium-speed ultrafiltration centrifugation on the supernatant collected in the step (3) at a temperature of 0-4 DEG C for multiple times, and performing concentrating to a certain volume to obtain the anaerobic ammonium oxidation bacteria extracellular protein. The extracellular protein with high content can be obtained, the biological activity of the protein is high, the cell dissolution rate is low, the cellrupture is small, and the structural composition is not damaged; and the extracellular protein can be directly used for subsequent deep proteomics analysis and detection.

Description

technical field [0001] The invention relates to the technical field of extraction of biofilm extracellular proteins, in particular to a method for extracting biofilm extracellular proteins with quartz sand and non-ionic surfactants. Background technique [0002] Anammox bacteria (Anammox) are a kind of chemoautotrophic microorganisms that grow strictly in anaerobic and light-proof habitats, belonging to Planctomycetes. Such "red bacteria" can directly convert ammonia nitrogen (NH 4+ -N) and nitrite (NO 2- -N) into nitrogen gas (N 2 ), greatly shortening the nitrogen cycle pathway. Therefore, the wastewater denitrification process based on anammox bacteria (Anammox) has become the most energy-saving environmental biotechnology at present. [0003] Anammox bacteria (Anammox) have various forms, but most of them exist in the form of biofilm. Anammox bacteria (Anammox) biofilm extracellular protein is the most important active substance in its extracellular polymer (EPS), a...

AI Technical Summary

Problems solved by technology

Chemical extraction (such as: acid-base extraction, CER) is simple and efficient, with a large amount of extraction, but it is very easy to cause cell lysis, destroy the active structure of extracellular proteins, and interfere with downstream analysis; even if subsequent purification operations such as dialysis and precipitation, it will Inevitable secondary loss of extracellular proteins

Biological enzymatic hydrolysis, fast and gentle reaction, easy to control, but its extraction cost is high, which is not conducive to large-scale industrial application production

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