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Method for quickly and simply analyzing microbial community structure of yeast for traditional brewage

A microbial community, a simple technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, analysis materials, etc., to achieve the effect of effective analysis methods, solving analysis errors, and simple operation

Inactive Publication Date: 2011-04-27
SICHUAN UNIV
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Problems solved by technology

However, using the improved phospholipid fatty acid analysis method described in this patent to analyze the microbial community structure of Daqu for traditional brewing, there has been no relevant literature or patent reports at home and abroad so far.

Method used

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  • Method for quickly and simply analyzing microbial community structure of yeast for traditional brewage

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Embodiment Construction

[0012] Add 3.2ml of phosphate buffer, 4ml of chloroform and 8ml of methanol to 3g of Daqu sample successively, shake in the dark at room temperature at 250r / min for 2h. Add 4.8ml of buffer solution and 6ml of chloroform successively, shake vigorously and mix well, and let stand overnight. The chloroform phase (lower layer) was transferred to a new test tube and blown dry with nitrogen. The extracted fatty acid was dissolved in chloroform, transferred to a silica gel column, and eluted with 10ml chloroform, 10ml acetone, and 10ml anhydrous methanol respectively, and the methanol eluate was collected and blown dry with nitrogen. Add 1ml of toluene:methanol ((1:1) v / v) to the dried solid component, 1ml of 0.2mol·l -1 Potassium hydroxide-methanol, 37°C water bath for 15min, after cooling, add 2ml ultrapure water and 0.3ml acetic acid solution (1mol / L), extract FAME with n-hexane (4ml), take the organic phase, blow dry with nitrogen, add internal standard FAME samples were analyz...

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Abstract

The invention relates to a culture-free biochemical method for analyzing the microbial community structure of yeast for traditional brewage, in particular to a method for analyzing the microbial community structure of yeast for traditional brewage by using the phospholipid fatty acid spectrogram analysis technique. The method comprises the following steps: (1) weighting a certain amount of yeast sample which is pulverized and evenly mixed; (2) adding a buffer solution and an organic reagent to obtain an organic phase containing lipids; (3) separating the organic phase by small silica gel columns, eluting and collecting organic components containing polar lipids; (4) carrying out methyl esterification under mild condition, and adding an extracting reagent to obtain corresponding fatty acid methyl ester; (5) carrying out quantitative analysis on the fatty acid methyl ester by using a gas chromatogram-mass spectrogram combined instrument; and (6) determining the characteristics of the microbial community structure in the sample according to characteristics of different phospholipid fatty acid compositions. The invention provides an accurate and quick culture-free detection means for researching the composition of the microbial community structure of yeast for brewage and knowing the law of growth and decline of different microbial communities in the yeast preparation process.

Description

technical field [0001] The invention belongs to the field of non-culture analysis of Daqu microbial community structure. Phospho lipid fatty acid (PLFA) is an important component of living cell membranes of almost all microorganisms (except archaea), and can be quickly quantified based on the analysis results of phospholipid fatty acids in Daqu samples. Analysis of the composition of complex microbial communities in Daqu. Background technique [0002] Daqu for brewing liquor, vinegar, etc. is not only an important biocatalyst in the brewing process, but also one of the main sources of aroma components in the product. The unique production process and long production history endow the microbial community structure with diversity and temporal and spatial characteristics. A large number of microbial resources adapted to extreme environments are also enriched in the production environment that has been domesticated for a long time. Based on the specific analysis of the microbi...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72C12Q1/04
Inventor 周荣清秦臻黄钧
Owner SICHUAN UNIV
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