Cellulose degradation composite microbial inoculum with low-temperature resistance as well as preparation method and application of cellulose degradation composite microbial inoculum

A technology for degrading cellulose and compound bacterial agents, which is applied in biochemical equipment and methods, preparation of organic fertilizers, and methods based on microorganisms, etc. The implementation process is complicated and other problems, to achieve the effect of easy implementation, efficient degradation, and simple preparation method

Pending Publication Date: 2019-04-09
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, people's research on cellulose-degrading bacteria mainly focuses on medium-high temperature degrading bacteria, and there are many commercial microbial straw-degrading bacteria agents on the market, and they are applied to straw returning to the field, but most of them are aimed at medium-high temperature or

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  • Cellulose degradation composite microbial inoculum with low-temperature resistance as well as preparation method and application of cellulose degradation composite microbial inoculum
  • Cellulose degradation composite microbial inoculum with low-temperature resistance as well as preparation method and application of cellulose degradation composite microbial inoculum
  • Cellulose degradation composite microbial inoculum with low-temperature resistance as well as preparation method and application of cellulose degradation composite microbial inoculum

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Embodiment 1

[0034] This embodiment is an exemplary preparation method of a low-temperature-resistant cellulose-degrading bacteria composite bacterial agent. The method of this embodiment includes the following steps:

[0035] 1) Take Flavobacterium johnsoniae and Stenotrophomonas maltophilia with inoculation loops respectively, numbered X24 and X26 respectively, and insert each loop strain into NA medium for activation and expansion culture in turn ;

[0036] 2) Pick the activated colonies and inoculate them in the corresponding liquid fermentation medium respectively, and culture them with shaking at 25°C and 200r / min for 48 hours to the logarithmic growth phase to obtain liquid bacterial agent seed liquid;

[0037] 3) After amplifying the above-mentioned seed solution to obtain a bacterial solution with a suitable concentration, collect the bacterial cells after high-speed centrifugation to prepare a dry powder bacterial agent, and mix it at a weight ratio of 1:1.

Embodiment 2

[0039] This example is the determination of the cellulase activity and degradation rate of the low temperature resistant cellulose degrading bacteria alone and the composite bacterial agent.

[0040] (1) Determination of enzyme activity

[0041] The two strains of single bacteria and the composite bacterial agent in Example 1 were respectively inserted into the enzyme-producing fermentation medium, and cultured at 12 ° C. Using straw powder as a carbon source, the two strains were continuously fermented for 5 days to produce enzymes. And measure FPA enzyme activity, CMC enzyme activity and β-glucosidase activity with DNS method every 24h, do 3 groups of parallels for each bacterial strain, measure the highest activity of each cellulase of 2 single strains and compound bacterial agents, such as figure 1 shown. Enzyme production medium composition: KH 2 PO 4 2g, (NH 4 ) 2 SO 4 1.4g, MgSO 4 ·7H 2 O0.3g, CaCl 2 0.3g, FeSO 4 ·7H 2 O 0.005g, MgSO 4 ·7H 2 O 0.0016g, Z...

Embodiment 3

[0048] This embodiment is a method for in-situ degradation of straw and returning it to the field. The composite bacterial agent adopted in the method of the present embodiment adopts the microbial agent of Example 1. Method of the present invention comprises the following steps:

[0049] 1) According to the dosage of 4.5kg of bacterial agent per mu of land, mix the bacterial agent with the soil and spread it on the straw;

[0050] 2) According to the dosage of 7kg urea per mu of land, mix urea with an appropriate amount of water and sprinkle it on the straw;

[0051] 3) Rotary tillage to bury the straw into the soil, keep the water content of the straw and soil at 50%-65%, and degrade the straw in a low temperature environment for ≥ 45 days, so that the straw is fully decomposed.

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Abstract

The invention discloses a cellulose degradation composite microbial inoculum with low-temperature resistance as well as a preparation method and an application of the cellulose composite microbial inoculum. The cellulose degradation composite microbial inoculum with low-temperature resistance comprises effective components of flavobacterium johnsoniae and stenotrophomonas maltophilia in a weight ratio of (0.5-1.51): 1, and the total effective viable count in the cellulose degradation composite microbial inoculum with low-temperature resistance is 3*10<9> CFU/g or more. The cellulose degradation composite microbial inoculum with low-temperature resistance can be further applied to in-situ degradation and returning to the field of t straws due to efficient degradation of cellulose such as crop straws at low temperature. The composite microbial inoculum can solve the problems of low cellulose degradation rate and long period in the low temperature environment in northern China.

Description

technical field [0001] The invention relates to the field of straw degradation, in particular to a low-temperature-resistant cellulose degradation composite bacterial agent, a preparation method and application thereof. Background technique [0002] Cellulose is a chain polymer composed of D-glucopyranose bonded by β-1,4-glucosidic bonds. It has a high-strength network structure and is insoluble in water and general organic solvents. As the most abundant natural organic renewable resource in nature, cellulose is also the main component of crop straw. Due to its stable physical and chemical properties, it is difficult to be decomposed under natural conditions and cannot be rationally utilized. The world's annual straw production is about 2.2 billion tons, and my country's annual crop straw production is about 800 million tons. A large number of crop straws are left idle or burned, which not only wastes resources, but also causes serious environmental pollution. Cellulase is ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/00C05F11/08C05F17/00C12R1/20C12R1/01
CPCC05F11/00C05F17/00C12N1/20C05F11/08Y02W30/40
Inventor 邹洪涛于素素叶旭红于双范庆锋张玉玲虞娜党秀丽王秋菊闫大波
Owner SHENYANG AGRI UNIV
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