High-temperature Daqu leaching liquor macro-protein extraction and purification method

Abstract

The invention relates to a high-temperature Daqu leaching liquor macro-protein extraction and purification method. The method comprises the following steps: crushing high-temperature Daqu, spraying sterile water and evenly mixing, storing in a biological culture tank, performing activated culturing for later use by adopting a stage temperature rise method; adding an acetic acid-sodium acetate buffer solution and a phenylmethylsulfonyl fluoride solution, shaking and blending evenly, and then leaching overnight, filtering and centrifuging to obtain high-temperature Daqu leaching liquor; sequentially adding a TCA acetone solution, an ammonium acetate methanol solution, an acetone solution, a Tris satured phenol / lauryl sodium sulfate buffer solution, an ammonium acetate methanol solution, and a methanol and acetone solution to obtain sediment A, sediment B, phenol layer, sediment C, sediment D and high-temperature Daqu macro-protein samples; adding a sample lysate, performing ice bath ultrasonic for hydrotropy, and centrifuging the sediments to obtain a high-temperature Daqu macro-protein sample solution. The high-temperature Daqu macro-protein sample solution prepared by adopting the method can be applied to two-dimensional electrophoresis of the high-temperature Daqu macro-protein, to obtain high-resolution and high-repeatability two-dimensional electrophoretogram.

Description

technical field [0001] The invention belongs to the technical field of macroproteomics. In particular, it relates to a method for extracting and purifying macroproteins from high-temperature Daqu extract. Background technique [0002] High-temperature Daqu is made of wheat as the main raw material. It is crushed and pressed into brick-shaped koji blocks. It is cultivated openly under certain temperature and humidity conditions. The highest temperature for koji-making cultivation is 60~68°C. The koji mother and the microorganisms in the raw materials grow, multiply and metabolize various enzymes on the koji block, which makes high-temperature Daqu have liquefaction, saccharification and protein decomposition capabilities. High-temperature Daqu is mainly used in the production of Maotai-flavored liquor, typically Moutai. The complex and various enzyme systems or enzyme protein systems produced by microorganisms in high-temperature Daqu are the main driving force for the form...

AI Technical Summary

Problems solved by technology

Aiming at the problem, the present invention uses high-temperature Daqu as material, conducts activation culture with high-temperature Daqu, improves the protein extraction method, basically solves the interference problem of melanoid pigments in high-temperature Daqu, and constructs a high-resolution, high-repeatability high-temperature Samples of macroproteins extracted from Daqu extract for high-resolution, high-reproducibility two-dimensional electrophoresis

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