Bacillus firmus and application thereof
A technology of bacillus firmus and bacteria agent, applied in the field of microorganisms, can solve the problems of natural ecosystems and human health threats, petroleum-based plastics are not biodegradable, and achieve good industrial application prospects
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Embodiment 1
[0042] Example 1 Isolation and Screening of Bacillus firmus MN36-4
[0043] This embodiment provides the preparation process of Bacillus firmus MN36-4, refer to figure 1 ,Specific steps are as follows:
[0044] 1. Isolation of culturable strains
[0045] Collect 10cm of soil from the tidal flat surface in the Mangrove Reserve, Futian District, Shenzhen; put the collected soil into a conical flask filled with 100mL of sterilized oil-containing and salt-enriched medium, and shake at a constant temperature of 30°C and 160r / min Cultivate for 77 days, transfer to the enriched medium containing fresh medium with 1% inoculum every 7 days, increase the oil content by 1% and salinity by 10‰ at the same time, take 1mL of soil on the 7th, 42nd and 77th days of cultivation Suspension; gradiently dilute the sediment suspension to make a soil suspension with a concentration of 10-3 to 10-5; add the diluted sediment suspension to a 2216E medium plate for coating treatment, 30°C After 48 h...
Embodiment 2
[0051] The identification of embodiment 2 Bacillus firmus MN36-4
[0052] This example identifies the Bacillus firmus MN36-4 obtained in Example 1, including morphological observation, physiological and biochemical identification, and 16S rDNA sequence analysis, as follows:
[0053] 1. Morphological observation
[0054] Streak the bacterial strain MN36-4 provided in Example 1 onto a 2216E medium plate, then invert the plate, culture it in a 30°C incubator for 24 hours, observe and record the growth of the colonies on the plate. Colony morphology reference for strain MN36-4 image 3 . from image 3 It can be seen that the colony of the strain is light yellow, oval, and flat, with irregular edges, slightly turbid, and a moist and smooth surface.
[0055] Carry out Gram staining to the bacterial strain MN36-4 provided by the embodiment of the invention with the kit, and observe bacterial strain under the oil lens, the Gram staining figure of this bacterial strain sees Figur...
Embodiment 3
[0069] Example 3 Determination of PHA-producing Ability of Bacillus firmus MN36-4
[0070] In the embodiment of the present invention, the PHA-producing ability of Bacillus firmus MN36-4 was measured, and the determination included the following contents:
[0071] 1. Extraction of PHA
[0072] The Bacillus firmus MN36-4 provided in Example 1 was inoculated into the liquid medium, and cultured with constant temperature shaking at 30°C and 150 r / min for 4 days. After the cultivation, the fermentation broth was centrifuged at 5000r / min for 20min to obtain cell pellets, and then freeze-dried; weighed 10mg of bacterial freeze-dried samples, put them into lipidation tubes, and then added 1mL of chloroform (containing 0.5mg / mL methyl benzoate) and 1mL methanol solution containing 15% (v / v) concentrated sulfuric acid, sealed in an oil bath at 100°C for 2.5h to carry out methylation reaction; after the reaction, cool the sample in an ice bath for 5min , then add 0.5mL deionized wate...
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