MethyLomonas tolerant to high-concentration methanol and application thereof
A methylomonas, high-concentration technology, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., to achieve the effect of convenient operation, simple formula and high industrial application value
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Embodiment 1
[0047] Screening method for Methanotroph MethyLomonas SP.ZR1:
[0048] The screening of Methylomonas comes from wetland sludge. Inoculate 1g of wetland sludge sample directly into a 250mL anaerobic bottle containing 100mL of NMS medium, and supplement 100mL of methane gas as a carbon source in the anaerobic bottle , so that the content of methane in the gas phase of the anaerobic cylinder reaches 30%. Culture at 25°C with shaking at 180 rpm until the OD600 of the culture medium reaches a stable level. 1% of the inoculum was inoculated in a fresh medium, and subcultured for 3 times. The liquid culture was diluted to coat the plate, and the plate was cultured in air containing 30% methane. A large number of methanotrophs can be screened by this screening method, among which Methylomonas ZR1, which forms bright orange-red colonies, grows the fastest and forms the largest colony.
[0049] The genome of ZR1 was extracted, and its 16S rRNA gene sequence was amplified and sequence...
Embodiment 2
[0051] Growth of MethyLomonas SP.ZR1 on methane as substrate:
[0052] A single colony of ZR1 formed on the plate was picked and inoculated in 30 mL of NMS liquid medium (methane as the carbon source) for culture at 28° C. and 180 rpm. Sampling and measuring the bacterial concentration of the fermentation broth every day, from such as figure 1 As shown, as the fermentation time prolongs, the cell concentration gradually increases, indicating that the number of cells of Methylomonas gradually increases, and this strain grows well in the medium with methane as the substrate.
Embodiment 3
[0054] Growth of MethyLomonas SP.ZR1 on Methanol as Substrate:
[0055] ZR1 grows a single colony of ZR using methanol as a substrate, inoculates it in 30 mL of NMS liquid medium (methane is the carbon source), and conducts seed culture at 28°C and 180 rpm. Cultured for 2 days, OD600 reached 0.7, inoculated in fresh methanol content of 12.5g / L, 15g / L, 17.5g / L, 20g / L, 22.5g / L, 25g / L, 27.5 g / L, 30g / L, 32.5g / L, 35g / L NMS medium 100mL, shake culture in 250mL shake flask, 25℃, 180rpm culture for 3 days, the culture results are as follows figure 2 As shown, it shows that Methylomonas can grow and reproduce well under the conditions of different concentrations of methanol.
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