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A separation method and microbial technology, applied in the field of microbial separation, can solve the problems of cumbersome operation, clean screening, and low efficiency, and achieve the effects of improving separation efficiency, preventing hyphae from balling, and accelerating the proliferation rate
Inactive Publication Date: 2020-06-23
HEBEI GUXIANGYANG WINE CO LTD
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At present, the method of heating in a water bath at 80°C is usually used to pretreat the pit mud samples, because the spores of caproic acid bacteria are heat-resistant, while their vegetative cells are not heat-resistant, weaker spores can be eliminated by heating, and at the same time Kill vegetative cells and other miscellaneous bacteria, but there are many kinds of m...
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Embodiment 1
[0022] Step ①: Collect the pit mud in the cellar pool of the ancient Xiangyang winery in Hubei Province with the five-point sampling method, quickly put the collected pit mud into an anaerobic bag, put it on ice and transport it to the laboratory;
[0023] Step ②: Weigh 10g of the pit mud sample, add it into a triangular flask filled with sterile water and glass beads, and vibrate in a rotary shaker to obtain a pit mud suspension;
[0024] Step ③: The pit mud suspension is heated in a water bath at 80°C for 10 minutes to kill the vegetative cells, and then inoculated in the enrichment medium, which is distiller's lees infusion medium, and the components contained are calculated as : 1000 parts of distiller's grain extract, 50 parts of loess, 10 parts of yeast extract, 5 parts of NaAc, 10 parts of ethanol, MgSO 4 ·7H 2 O 0.3 parts, CaCO 3 5 parts, 10 parts of polyglutamic acid, 5 parts of sucrose ester, 37 ° C enrichment culture for 6 days, select the enrichment medium with e...
Embodiment 2
[0029] Embodiment 2 differs from Embodiment 1 in that:
[0030] Distiller's grain extract medium, the components contained in parts by mass are: 1000 parts of distiller's grain extract, 150 parts of loess, 10 parts of yeast extract, 3 parts of NaAc, 20 parts of ethanol, MgSO 4 ·7H 2 O 0.1 part, CaCO 3 5 parts, 20 parts of polyglutamic acid, 10 parts of sucrose ester, 37 ° C enrichment culture for 4 days.
Embodiment 3
[0032] Embodiment 3 differs from Embodiment 1 in that:
[0033] Distiller's grain extract medium, the components contained in parts by mass are: 1000 parts of distiller's grain extract, 50 parts of loess, 10 parts of yeast extract, 5 parts of NaAc, 10 parts of ethanol, MgSO 4 ·7H 2 O 0.3 parts, CaCO 3 5 parts, 30 parts of polyglutamic acid and 15 parts of sucrose ester were enriched and cultured at 37°C for 4 days.
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Abstract
The preset invention belongs to the technical field of microorganism separation and discloses a pit mud microorganism separation method. The pit mud microorganism separation method comprises the following steps: step 1, collecting pit mud by a five-point sampling method; step 2, weighing 10 g of a pit mud sample to prepare a pit mud suspension; step 3, after water bath, inoculating the pit mud suspension into an enrichment culture medium, coating the enrichment culture medium into a separation culture medium by adopting a gradient dilution manner, placing the inoculated separation cultured medium into an anaerobic tank, performing separation culture, and performing plate streaking purification on microorganism colonies with various different characteristics to obtain single microorganism colonies; and step 4, streaking and separating the obtained single microorganism colonies and conducting caproic acid bacteria identification of the separated strain. Vinasse immersion liquid is used as the culture medium to simulate primary growth environment of microorganisms, so that the microorganisms in the pit mud can rapidly adapt to the new environment, activation time of the microorganismsis shortened, and microorganism separation efficiency can be improved.
Description
technical field [0001] The invention belongs to the technical field of microorganism separation, and in particular relates to a method for separating microorganisms from pit mud. Background technique [0002] To produce Luzhou-flavor wine, pit mud is the foundation, Daqu is the driving force, and craftsmanship is the key. To make Luzhou-flavored high-quality wine, the quality of pit mud must first be grasped. The quality of pit mud directly determines the quality of wine. Because pit mud is the carrier and habitat of caproic acid bacteria, methane bacteria, butyric acid bacteria and other beneficial substances, as well as its breeding ground, the type and quantity of these beneficial microorganisms are a standard to measure the quality of pit mud. The main aroma substance of Luzhou-flavor Daqu liquor is ethyl caproate, and ethyl caproate is a metabolite of various aroma-producing and ester-producing microorganisms such as Clostridium (caproic acid bacteria) in the pit mud,...
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