Bacillus licheniformis for straw degradation and application thereof
A technology of Bacillus licheniformis and straw, which is applied to microorganism-based methods, bacteria, fungi, etc., can solve the problems of environmental pollution and low utilization rate of straw, achieve strong tolerance, improve maturity and fertilizer efficiency, and improve maturity. degree of effect
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Embodiment 1
[0033] Example 1 strain screening
[0034] 1.1 Sample:
[0035] Collected from broiler waste in Penglai, Yantai.
[0036] 1.2 Initial screening of cellulose-degrading strains:
[0037] Take 1g of chicken manure and add it to a culture bottle containing 99 mL of sterile water, and add a few glass beads to make a suspension; take 1 mL of the suspension and put it into a culture bottle containing 50 mL of enriched medium, seal it , placed in a biochemical incubator at 30°C for static culture for 3-7 days. Spread the enriched culture on the separation medium, culture at 30°C for 2 days, pick out a single colony for purification, and then continue to purify on the separation medium for 2 to 3 times.
[0038] Finally, the applicant successfully isolated three strains with cellulose-degrading ability by using the enrichment medium and separation medium with carboxymethylcellulose sodium (CMC-Na) as the sole carbon source, and named them VB372, VB375, VB376.
[0039] 1.3 Re-scree...
Embodiment 2
[0056] The identification of embodiment 2 VB376 bacterial strain
[0057] 2.1 Identification of colony morphology
[0058] The colony morphology of the VB376 strain is as follows figure 1 As shown, the colony is off-white, 3-5mm in diameter, with rough edges and a bulge in the middle. Electron microscope observation results such as figure 2 As shown, the VB376 bacterium is a short straight rod-shaped, Gram-positive bacterium that can produce spores, the sporangia do not expand, and the cells are solitary.
[0059] 2.2 16S rDNA molecular identification
[0060] The genome of the VB376 strain was extracted using a kit. Then using the genome as a template, the 16S rDNA was amplified using specific primers. The amplified PCR products were detected by 1% agarose gel electrophoresis and sent to a sequencing company for sequencing.
[0061] Sequencing results showed that the sequence of the PCR amplification product was SEQ ID NO:1. Through the BLAST comparison of the sequen...
Embodiment 3
[0073] Example 3 Evaluation of Enzyme Production Ability and Low Temperature Resistance of Bacillus licheniformis VB376
[0074] 1. Preparation of bacteria solution
[0075] The activated Bacillus licheniformis VB376 was inoculated into LB liquid medium, cultivated at 37°C and 220rpm for 14h, and the viable count was 10 8 -10 9 CFU / ml bacterial solution, adjusted to 10 with sterile LB medium 8 CFU / ml.
[0076] 2. Evaluation of cellulase production ability
[0077] Spot the Bacillus licheniformis VB376 bacteria solution on the separation medium, and incubate at 37°C for 48 hours. A transparent circle can be seen around the Bacillus licheniformis VB376, and the diameter of the transparent circle is 18 mm. This shows that Bacillus licheniformis VB376 can produce certain cellulase.
[0078] 3. Evaluation of the ability to produce endoglucanase
[0079]The Bacillus licheniformis VB376 bacterial liquid was centrifuged at 4°C and 12000 rpm for 10 min, and the enzyme activity of...
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