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A strain of Bacillus licheniformis producing cellulase and its application

A technology of Bacillus licheniformis and cellulase, which is applied in the field of microorganisms, can solve the problems of large labor input, insignificant treatment effect of microbial agents, and influence on farmers' enthusiasm, etc., and achieves long shelf life, broad market application prospects, easy transportation and stored effect

Active Publication Date: 2019-10-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the main method of returning straw to the field is to collect and crush the straw and pile it up in the field, supplement urea and other substances appropriately, mix in microbial agents, and stir at the right time. Using this method requires a lot of labor and affects the enthusiasm of farmers.
The most direct way is to treat the straw without processing and in situ. However, the straw is usually returned to the field in autumn and winter. Due to the low temperature, the treatment effect of various existing microbial agents is not significant.

Method used

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  • A strain of Bacillus licheniformis producing cellulase and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Screening of Bacillus licheniformis CCTCC No.M 2016202

[0024] Collect 1g of soil sample from a paddy field in Wuxi, Jiangsu, add it to 99ml of sterile saline, shake it on a shaker at 37°C for 10min, and the shaker speed is 150rpm; absorb 1ml of liquid and add CMC (sodium carboxymethylcellulose) as the only carbon In the liquid medium of the source, continue shaking culture at 37°C and 150rpm; after 24 hours, draw 1ml of the liquid and add it to the liquid medium with CMC (sodium carboxymethylcellulose) as the only carbon source, at 37°C, Continue shaking culture at 150rpm for 24hrs; after 24hrs, absorb 1ml of liquid and add it to the liquid medium with CMC (sodium carboxymethylcellulose) as the sole carbon source, and continue shaking culture at 37°C and 150rpm for 24hrs. This was repeated three times for enrichment culture. Finally, after gradient dilution of the culture solution, spread on a plate medium with CMC (sodium carboxymethylcellulose) as the sol...

Embodiment 2

[0027] Example 2 Identification of Bacillus licheniformis CCTCC No.M 2016202

[0028] 1. Colony morphology

[0029] After the strain was cultured on LB medium at 37°C for 24 hrs, round colonies were formed with irregular edges; Gram staining was positive; under optical microscope observation, the bacterial cells were long rod-shaped, single, paired or chain-like.

[0030] 2. Physiological and biochemical identification

[0031] Physiological and biochemical identification is carried out to the screened bacterial strain in embodiment 1, and the results are as shown in table 1:

[0032] Table 1 Physiological and biochemical identification results of cellulase-producing strains

[0033] project result project result Athletic + Oxidase + spore + Methyl red + Grow at 37°C + V-P determination + anaerobic growth + indole production - glucose + Catalase + citrate utilization + propionate utilization + ...

Embodiment 3

[0037] Example 3 Determination of Cellulase Production Ability of Bacillus licheniformis CCTCC No.M 2016202

[0038] Cellulase production test medium (g / L): maltose 40, peptone 10, NaCl 5, K 2 HPO 42. Microbial culture conditions: liquid volume 15mL / 250mL Erlenmeyer flask, shaker temperature control 37°C, rotation speed 180rpm, culture time 48hr. Determination of cellulase activity by CMC method (Wu Wentao, Ju Meiting, Liu Jinpeng, etc., Isolation, Identification and Degradation Characteristics of a Cellulose Degrading Bacteria Strain to Corn Stalk. Microbiology Bulletin, 2013, 40(4): 712-719) . Bacillus licheniformis CCTCC No.M 2016202 was used as the test bacteria, and Bacillus licheniformis CCTCC No.M 2012458 was used as the control bacteria.

[0039] After measurement, the enzyme activity of Bacillus licheniformis CCTCC No.M 2016202 fermentation liquid is 98.5U / mL, and the enzyme activity of control bacterial strain Bacillus licheniformis CCTCC No.M 2012458 fermentatio...

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Abstract

The invention relates to a cellulase-producing bacillus licheniformis, a microecological preparation containing the bacillus licheniformis, and an application of the microecological preparation in promoting straw maturity, belonging to the technical field of microorganisms. The strain has been preserved in the China Center for Type Culture Collection on April 18, 2016. The strain preservation number is CCTCC No.M 2016202. It uses liquid fermentation, centrifugation, and spray drying to obtain a single bacterial powder, and Bacillus subtilis CCTCC No.M 2011443, Bacillus coagulans CCTCC No.M 2015273, and Aspergillus niger are mixed in a certain proportion, and auxiliary materials are added to prepare the straw decomposing agent. The straw decomposing agent added with Bacillus licheniformis CCTCC No.M 2016202 of the present invention can degrade the crop straws in situ and promote the returning of the straws to the field, especially in autumn and winter when the temperature is low.

Description

technical field [0001] The invention relates to a cellulase-producing bacillus licheniformis, a microecological preparation containing the bacillus licheniformis, and an application of the microecological preparation in promoting straw decomposing, belonging to the technical field of microorganisms. Background technique [0002] The annual production of crop straw in China is more than 700 million tons, and only a small part is used in feed, building materials, power generation, fuel ethanol and other fields; most of them are burned on site, which not only causes fire hazards, but also pollutes the air environment. [0003] In fact, straw is rich in organic matter and effective nutrients such as nitrogen, phosphorus, and potassium. Returning straw to the field can increase soil organic matter, enhance soil water retention capacity, and prevent soil erosion. A large number of studies have shown that after returning straw to the field, the soil quality is significantly improve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/14C05F11/00C05F17/00C12R1/10C12R1/125C12R1/07C12R1/685
CPCC05F11/00C05F17/00C12N1/14C12N1/20C12N1/205C12R2001/10C05F11/08Y02W30/40
Inventor 张梁李颜颜顾正华丁重阳李由然李赢石贵阳
Owner JIANGNAN UNIV
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