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

A high-temperature Daqu and purification method technology, applied in the field of metaproteomics, can solve the problems of melanoid pigment interference, etc., and achieve the effect of high repetition rate, clear background and clear resolution

Active Publication Date: 2016-11-09
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 mela...

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

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Embodiment 1

[0035] In order to realize the purpose of the present invention, the technical scheme of adopting high-temperature Daqu extract to extract macroprotein is as follows:

[0036] 1. Activation culture of high temperature Daqu:

[0037] On the koji block of high-temperature Daqu, take 300g of high-temperature Daqu according to the five-point sampling method, pulverize it and pass it through a 40-mesh sieve to obtain high-temperature Daqu powder, spray 40g of sterile water and mix well, and place it in a biological incubator for 24-hour activation culture. That is, activate and cultivate at 30°C for 8 hours, activate at 35°C for 6 hours, activate at 40°C for 6 hours, and activate at 45°C for 4 hours before use.

[0038] 2. Preparation of high-temperature Daqu extract:

[0039] Weigh 30g of activated high-temperature Daqu powder, add 90ml of acetic acid-sodium acetate buffer solution with a pH value of 4.6 and 90μL of phenylmethylsulfonyl fluoride solution (PMSF solution), shake an...

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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...

Claims

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Application Information

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IPC IPC(8): C07K1/30
CPCC07K1/30
Inventor 黄镇黄祖新刘龙山
Owner FUJIAN NORMAL UNIV
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