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Pit mud flora enrichment method based on oligoculture and application thereof

A basic culture medium and pit mud technology, applied in the direction of biochemical equipment and methods, bacteria, alcoholic beverage preparation, etc., can solve problems such as hindering cultivability, lack of directional culture methods, and unknown nutritional requirements, so as to avoid production safety risk effect

Pending Publication Date: 2020-08-07
JIANGNAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for syntrophic bacteria and methanogens, there is a lack of effective directional culture methods
Syntrophic bacteria in pit mud hinder their culturability due to unknown nutritional requirements
As for the methane bacteria, because of their slow growth, the enrichment in the pit mud is relatively slow, and a certain number of methane bacteria can be obtained by continuous brewing for several years in the new pit pit; on the other hand, the methane bacteria are strict anaerobic bacteria , are difficult to be cultivated, and there is still a lack of cultivable techniques for typing and cultivating hydrogenotrophic and acetic acidotrophic methanogens
In addition, the current research has found that there are at least 100 kinds of prokaryotic microorganisms in the pit mud, and the uncultured bacteria are not limited to syntrophic bacteria and methanogens, but other bacterial types widely distributed in the pit mud, such as Aminobacterium, Hydrogen Bacteria (Hydrogenispora), Zymomonas (Fermentimonas), etc., have been difficult to cultivate even at the flora level

Method used

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  • Pit mud flora enrichment method based on oligoculture and application thereof
  • Pit mud flora enrichment method based on oligoculture and application thereof
  • Pit mud flora enrichment method based on oligoculture and application thereof

Examples

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Effect test

Embodiment 1

[0051] The present embodiment provides a method for enriching and culturing the acetic acid-trophic methane flora in the pit mud, and the method includes the following contents:

[0052] (1) Selection of pit mud samples: select 10-year-old pit bottom mud samples 1 and 2 containing acetic acidotrophic methane flora, the pH value of sample 1 pit mud is 6.5, and the pH value of sample 2 pit mud is 7.0. Anaerobic sampling bags were used to sample 100 g of fresh pit mud respectively, and the bacterial group structure analysis and inoculation enrichment culture were carried out.

[0053] (2) Preparation of medium: the enrichment medium used was composed of basal medium with sodium acetate trihydrate added, and the dosage of sodium acetate trihydrate was 8.6 g / L. The basic medium components are: mineral element solution 5.0% (v / v), trace element solution 0.1% (v / v), B vitamin solution 0.5% (v / v), yeast powder 1g / L, peptone 1g / L , NaHCO 3 Solution 7.0% (v / v), Na 2 S·9H 2 O in wat...

Embodiment 2

[0068] This embodiment provides a method for enriching and culturing the syntrophomonas group in the pit mud, and the method includes the following contents:

[0069] (1) Selection of pit mud samples: select a 5-year-old pit bottom mud sample containing syntrophomonas flora, the pH value of the pit mud is 6.0, and the butyric acid content is less than 5g / kg. Anaerobic sampling bags were used to sample 100 g of fresh pit mud respectively, and the bacterial group structure analysis and inoculation enrichment culture were carried out.

[0070] (2) Preparation of culture medium: the enrichment medium used is composed of basal medium with sodium butyrate added, the concentration of sodium butyrate is 55mmol / L, other components and their preparation methods are the same as in Example 1.

[0071] (3) Culture method: 50mL of the above-mentioned culture medium is divided into 100mL saline bottles, the selected seed pit mud is inoculated with 10% (m / v) inoculum, and enrichment culture i...

Embodiment 3

[0080] This embodiment provides a method for co-enriching and cultivating syntrophomonas and methane flora in pit mud, and the method includes the following contents:

[0081] (1) Selection of pit mud samples: 5-year-old pit bottom mud containing syntrophomonas and methane flora was selected, and the pH value of the pit mud was 6.5. Anaerobic sampling bags were used to sample 100 g of fresh pit mud respectively, and the bacterial group structure analysis and inoculation enrichment culture were carried out.

[0082] (2) Preparation of medium: the enrichment medium used is composed of sodium butyrate and sodium formate added to the basal medium, the concentration of sodium butyrate is 55mmol / L, the concentration of sodium formate is 24mmol / L, other components and their The configuration method is the same as that in Embodiment 1.

[0083] (3) Culture method: 50mL of the above-mentioned culture medium is divided into 100mL saline bottles, the selected seed pit mud is inoculated ...

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Abstract

The invention relates to a pit mud flora enrichment method based on oligoculture and application thereof. The pit mud flora enrichment method based on oligoculture comprises the following steps of: inoculating pit mud into an enrichment medium for culture to enrich pit mud flora; wherein the enrichment medium comprises a basic medium and a selective carbon source, the basic medium comprises mineral element substances, trace element substances, B vitamins, yeast powder, peptone, sodium bicarbonate, sodium sulfide and water, the selective carbon source comprises short-chain fatty acid and / or short-chain fatty acid salt. The enrichment method can perform culturable enrichment on various uncultured flora, can be applied to preparation of methane bacteria or butyric acid utilization flora bacterial liquid, methane flora-butyric acid utilization flora co-culture, butyric acid utilization bacteria or methane bacteria activity detection, artificial pit mud manufacturing and maintenance and thelike, and provides a leading technology for pit mud functional microorganism resource mining.

Description

technical field [0001] The invention belongs to the technical field of liquor brewing, and relates to a method for enriching cellar mud flora and its application, in particular to a method for enriching pit mud flora based on oligoculture and its application. Background technique [0002] There are twelve major flavor types of liquor in my country, which can be divided into pit mud-dependent type and non-pit mud dependent type according to whether there is pit mud in the brewing process. Liquor brewed with pit mud dependence mainly includes eight categories: Luzhou-flavored, sauce-flavored, phoenix-flavored, sesame-flavored, mixed-flavored, special-flavored, medicinal-flavored, and rich-flavored. Liquor mainly includes four types: Fen-flavored, Laobaigan-flavored, Rice-flavored, and Soy-flavored. [0003] For the pit mud-dependent liquor brewing system, the main microorganisms inhabiting the pit mud are strictly anaerobic bacteria, including bacteria and archaea. Among the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04C12G3/02
CPCC12N1/20C12Q1/04C12G3/02
Inventor 任聪徐岩吴建峰季方
Owner JIANGNAN UNIV
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