Preparation and application of a compound microbial agent for degrading garden waste

Abstract

The invention discloses a compound microbial agent with a function of effectively degrading cellulose at high temperature. The compound microbial agent contains five actinomycetes: Streptomyces thermoviolaceus, Streptomyces thermodiastaticus, Streptomyces thermocarboxydus, Streptomyces albidoflavus and Streptomyces thermovulgaris, and one bacteria: Brevibacillus borstelensis. The compound microbial agent solves the problem of slow composting and long cycle of garden waste due to the limitation in cellulose degradation difficulty, accelerates cellulose degradation in the composting process, shortens the fermentation period, and improves the conversion efficiency, thereby having a broad application prospect in the aspects such as the application of composting treatment and resource utilization of garden waste, and the like.

Description

technical field

[0001] The invention belongs to the field of environmental biology technology, and in particular relates to the preparation and application of a compound microbial agent for degrading cellulose by high temperature of garden waste. Background technique

[0002] Garden waste refers to the dead branches, fallen leaves, grass clippings, residual flowers, tree and shrub pruning and other plant residues produced by the natural fall of garden plants or artificial pruning. The main components are refractory cellulose and hemicellulose. In recent years, my country's garden waste has been increasing at an annual rate of 8%-10%, which has brought great resistance to the process of urban greening. At present, the main methods of dealing with garden waste in my country are incineration and filling. These two processing methods are not only wasteful The use of renewable resources has also caused serious environmental pollution. The use of biocomposting to degrade garden wa...

Examples

Embodiment 1

[0023] Example 1 Screening and identification of high-temperature cellulose-degrading bacteria

[0024] Samples were collected from composting of garden waste in an apple orchard in Changping District, Beijing at the early stage of high temperature, middle stage of high temperature, and late stage of high temperature material, and the accumulation of landscaping waste in the afforestation area of ​​Yanqing Plain in Beijing; 10 g of the above fresh samples were weighed and placed in a container containing 10 glass beads. , and filled with 90 ml of sterile water in a conical flask, placed in a shaker at 30°C and 150 rpm for 30 minutes to fully disperse the samples, and left to enrich at 50°C for 24 hours. Use a sterile pipette to draw 1 ml of the supernatant and add it to a test tube containing 9 ml of sterile water, which is 10 -1 Sample diluent, again from 10 -1 Take 1ml of the sample and add it to 9ml sterile water, which is 10 -2 sample diluent, and so on, to get 10 -3 、1...

Embodiment 2

[0027] Example 2 Antagonism between bacterial strains

[0028] The above-mentioned six cellulose-degrading strains were streaked two by two on the carboxymethyl cellulose medium but not intersecting each other, cultured at 50°C for 3 days. If a sterile zone is formed at the intersection of two bacteria, it means that a zone of inhibition has formed between the two bacteria, and the two bacteria are antagonistic to each other and cannot be combined together. combination. From the results, there is no antagonistic effect between the various strains, and they will not interfere with each other and affect the growth of the strains, so it can be made into a compound bacterial agent.

Embodiment 3

[0029] Example 3 Cellulase-producing Ability of Mixed Bacteria

[0030] The 6 strains of high-temperature and high-efficiency cellulose-degrading bacteria screened were inoculated in CMC-Na liquid medium, and cultured in a constant temperature shaker at 50°C with a rotation speed of 150rpm / min. Streptomyces, Streptomyces vulgaris and Brevibacillus Potsdam were cultured for 12 hours, Streptomyces carbomonoxide and Streptomyces albiflous were cultured for 72 hours, and the turbidity OD600 of each culture solution was adjusted to 0.2, and then the suspensions of these 6 strains were prepared according to the The ratio of volume to volume is evenly mixed to obtain a mixed bacterial liquid. Use the same inoculation amount of the mixed bacteria liquid and each single strain to carry out fermentation and enzyme production under the same fermentation conditions, and compare the enzyme production ability of the mixed bacteria group and the single strain, see figure 1 . Depend on f...

technical field

[0001] The invention belongs to the field of environmental biology technology, and in particular relates to the preparation and application of a compound microbial agent for degrading cellulose by high temperature of garden waste. Background technique

[0002] Garden waste refers to the dead branches, fallen leaves, grass clippings, residual flowers, tree and shrub pruning and other plant residues produced by the natural fall of garden plants or artificial pruning. The main components are refractory cellulose and hemicellulose. In recent years, my country's garden waste has been increasing at an annual rate of 8%-10%, which has brought great resistance to the process of urban greening. At present, the main methods of dealing with garden waste in my country are incineration and filling. These two processing methods are not only wasteful The use of renewable resources has also caused serious environmental pollution. The use of biocomposting to degrade garden wa...

Examples

Embodiment 1

[0023] Example 1 Screening and identification of high-temperature cellulose-degrading bacteria

[0024] Samples were collected from composting of garden waste in an apple orchard in Changping District, Beijing at the early stage of high temperature, middle stage of high temperature, and late stage of high temperature material, and the accumulation of landscaping waste in the afforestation area of ​​Yanqing Plain in Beijing; 10 g of the above fresh samples were weighed and placed in a container containing 10 glass beads. , and filled with 90 ml of sterile water in a conical flask, placed in a shaker at 30°C and 150 rpm for 30 minutes to fully disperse the samples, and left to enrich at 50°C for 24 hours. Use a sterile pipette to draw 1 ml of the supernatant and add it to a test tube containing 9 ml of sterile water, which is 10 -1 Sample diluent, again from 10 -1 Take 1ml of the sample and add it to 9ml sterile water, which is 10 -2 sample diluent, and so on, to get 10 -3 、1...

Embodiment 2

[0027] Example 2 Antagonism between bacterial strains

[0028] The above-mentioned six cellulose-degrading strains were streaked two by two on the carboxymethyl cellulose medium but not intersecting each other, cultured at 50°C for 3 days. If a sterile zone is formed at the intersection of two bacteria, it means that a zone of inhibition has formed between the two bacteria, and the two bacteria are antagonistic to each other and cannot be combined together. combination. From the results, there is no antagonistic effect between the various strains, and they will not interfere with each other and affect the growth of the strains, so it can be made into a compound bacterial agent.

Embodiment 3

[0029] Example 3 Cellulase-producing Ability of Mixed Bacteria

[0030] The 6 strains of high-temperature and high-efficiency cellulose-degrading bacteria screened were inoculated in CMC-Na liquid medium, and cultured in a constant temperature shaker at 50°C with a rotation speed of 150rpm / min. Streptomyces, Streptomyces vulgaris and Brevibacillus Potsdam were cultured for 12 hours, Streptomyces carbomonoxide and Streptomyces albiflous were cultured for 72 hours, and the turbidity OD600 of each culture solution was adjusted to 0.2, and then the suspensions of these 6 strains were prepared according to the The ratio of volume to volume is evenly mixed to obtain a mixed bacterial liquid. Use the same inoculation amount of the mixed bacteria liquid and each single strain to carry out fermentation and enzyme production under the same fermentation conditions, and compare the enzyme production ability of the mixed bacteria group and the single strain, see figure 1 . Depend on f...

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