Geobacterium galactose BWTGW1.1 and application thereof
A galactose and geological technology, applied in the field of environmental microorganisms, can solve the problems of harmlessness and low fertility, long time-consuming natural fermentation, serious odor pollution, etc., achieve great application potential, promote the fermentation process and the duration of the high-temperature stage , the effect of strong growth adaptability
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Embodiment 1
[0029] Example 1 Isolation, purification and preservation of Geobacter galactosus BWTGW1.1
[0030] In March 2020, three mid-layer samples were collected from Dongguan City Center Designated Slaughterhouse Co., Ltd. during the high-temperature stage (50°C) of composting and fermentation of pig manure and sludge organic waste.
[0031] Isolation, purification and preservation of high-temperature bacteria: Weigh 10 g of the collected three samples and suspend them in 50 mL of PBS (pH 7.4, concentration 0.2 mol / L), shake vigorously for 5 min, centrifuge at 1000 r / min for 3 times, each time for 5 min, and collect the supernatant to the centrifuge tube. Dilute the supernatant to different concentrations, take 10 -3 、10 -4 、10 -5 Gradient bacterial solution was spread on LB agar plate and incubated at 60°C for 48 hours. Pick the strains with better growth and larger colonies, continue to streak and separate on the LB agar plate, and purify repeatedly.
[0032] The purified colo...
Embodiment 2
[0033] Example 2 Identification of 16S rDNA of Geobacter galactosus BWTGW1.1
[0034] (1) Morphological identification: Spread the screened bacterial strain BWTGW1.1 on an LB plate, culture at 60°C for 24 hours, and observe its colony characteristics; take a single colony smear, Gram staining, and observe its cell morphology under a microscope , see the result figure 1 .
[0035] (2) Molecular identification of 16S rRNA gene:
[0036]Genomic DNA of strain BWTGW1.1 was extracted, and the PCR product was amplified with bacterial 16S rDNA gene amplification general primer 27F / 1492R (5'-AGAGTTTGATCCTGGCTCAG-3' and 5'-TACGACTTAACCCCAATCGC-3') and sent to Shanghai Meiji Biomedical Technology Co., Ltd. (Guangzhou Branch) performed sequence sequencing, and the length of the 16S rRNA sequence obtained after sequencing was 1447bp, and the sequence is shown in SEQ ID NO.1. The sequencing results were compared with the 16SrDNA sequences in the NCBI and EzBioCloud databases for homology...
Embodiment 3
[0039] Example 3 Evaluation of the ability to produce thermostable protease of Geobacter galactosus BWTGW1.1
[0040] The ability to produce high-temperature-resistant protease was evaluated by the plate hydrolysis circle detection method:
[0041] The main media and reagents used are as follows:
[0042] Protein culture medium: 40.0g skimmed milk powder, 10.0g soluble starch, 3.0g yeast extract, 2.0g potato flour, 15.0g agar, add distilled water to 1000mL, pH 7.0-7.2. Sterilize at 121℃ for 20min at high temperature and high pressure, pour plate .
[0043] Spot the activated Geobacter galactosus BWTGW1.1 on the protein medium plate respectively, place them in a 60°C incubator and cultivate them for 48 hours, and observe whether there is a hydrolyzed transparent circle around the colonies, which can be directly reflected qualitatively. Whether the strain has the ability to secrete and produce thermostable protease.
[0044] Observation obtained by the above-mentioned method ...
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