High-density cultured nitrogen-fixing bacteria and preparing method and application thereof

A high-density cultivation and nitrogen-fixing bacteria technology, applied in the field of bioengineering, can solve the problems of low bacterial concentration and easy pollution, and achieve the effect of increasing the bacterial density

Inactive Publication Date: 2016-12-14
HEHAI TECHNOLOGY ENGINEERING GROUP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Existing high-density culture technology is usually obtained through interception or continuous culture wit

Method used

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  • High-density cultured nitrogen-fixing bacteria and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Such as figure 1 As shown, a method for high-density cultivation of nitrogen-fixing bacteria specifically includes the steps:

[0042] (1) Test tube expansion culture: Inoculate a ring of Azotobacter phusticosa slant strains into the nitrogen-fixing bacteria test tube slant medium, cultivate at 25°C for 10 days to obtain test tube slant strains, and store the test tube slant at 4°C for later use; The test tube slant strain is the general nutrient agar medium used;

[0043] (2) Liquid shake flask culture: Inoculate the slant strain of the test tube prepared in step B1 of the first ring into a 250mL Erlenmeyer flask with 20mL of liquid shaker culture medium. Under the conditions of shaking table culture for 86 hours, nitrogen-fixing bacteria liquid shake flask strains were made; the liquid shake flask culture medium was sterilized at 100°C for 60 minutes, and each liter of liquid shake flask culture medium contained sucrose 5g, peptone 1g, bran 2g, beef extract 1g, slud...

Embodiment 2

[0050] Such as figure 1 As shown, a method for high-density cultivation of nitrogen-fixing bacteria specifically includes the steps:

[0051] (1) Test tube expansion culture: Inoculate a ring of endogenous nitrogen-fixing bacteria slant strains into the nitrogen-fixing bacteria test tube slant medium, cultivate at 40°C for 2 days to obtain test tube slant strains, and store the test tube slant at 4°C for later use; The test tube slant strain is a general nitrogen-fixing bacterial culture medium;

[0052] (2) Liquid shake flask culture: Inoculate the slant strain of the test tube prepared in step B1 of the first ring into a 250mL conical flask with 150mL liquid shake flask culture medium. Under the conditions of shaking table culture for 18 hours, nitrogen-fixing bacteria liquid shake flask strains were made; the liquid shake flask culture medium was sterilized at 130°C for 10 minutes, and each liter of liquid shake flask culture medium contained sucrose 20g, peptone 10g, bra...

Embodiment 3

[0059] Such as figure 1 As shown, a method for high-density cultivation of nitrogen-fixing bacteria specifically includes the steps:

[0060] (1) Test tube expansion culture: inoculate a soybean rhizobium slant strain into the nitrogen-fixing bacteria test tube slant medium, cultivate for 6 days at 32° C. to obtain the test tube slant strain, and store the test tube slant at 4° C. for subsequent use; the test tube slant The strain is the general nitrogen-fixing bacteria medium used;

[0061] (2) Liquid shake flask culture: Inoculate the slant strains of the test tube prepared in step B1 of the first ring into a 250mL Erlenmeyer flask with 80mL of liquid shaker culture medium. Under the conditions of shaking table culture for 52 hours, nitrogen-fixing bacteria liquid shake flask strains were made; the liquid shake flask culture medium was sterilized at 120°C for 30 minutes, and each liter of liquid shake flask culture medium contained sucrose 12g, peptone 6g, bran 11g, beef e...

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Abstract

The invention discloses a preparing method and application of high-density cultured nitrogen-fixing bacteria, and belongs to the technical field of bioengineering. The culture method comprises the steps that biochar serves as an adsorbent, nitrogen-fixing bacteria serve as an original strain, test tube expansion culture, liquid shake flask culture, seed tank expansion culture and liquid fermentation culture are carried out in sequence, and the average thallus density of the cultured nitrogen-fixing bacteria reaches 5*1013-1*1015 cfu/mL. Obtained fermentation liquid can be subjected to centrifugation and spray drying, and the quantity of viable bacteria in dried thallus powder reaches up to 1015-1017 cfu/g. The obtained nitrogen-fixing bacteria can serve as a live bacteria agent of bio-fertilizer or as a live bacteria agent of bio-fertilizer together with other strains.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for cultivating nitrogen-fixing bacteria at high density and its application. Background technique [0002] The so-called biochar is made of organic waste, such as animal manure, animal bones, plant roots, wood chips and wheat straw, which are decomposed by high temperature through thermochemical effects, and become carbon-like spherical substances that can be used as fertilizers. Among them, the charcoal formed by plants for the purpose of fixing carbon is called "biological carbon". There is a large amount of carbon naturally present in the land, but this carbon is unstable and will release carbon dioxide due to the impact of climate. Biochar, on the other hand, can fix carbon for hundreds of years. [0003] People living in the Brazilian Amazon have long used a special kind of fertilizer. This fertilizer is locally sourced and has a strong ability to restore...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/08C12R1/065
CPCC12N1/20C05F11/08
Inventor 俞建国
Owner HEHAI TECHNOLOGY ENGINEERING GROUP CO LTD
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