Clostridium sporogenes with function of synchronously producing hydrogen by fermentation and methylating anaerobic arsenic and application of clostridium sporogenes
A technology of Clostridium sporogenes and methylation, applied in the field of microorganisms, can solve problems such as high energy consumption and carbon emissions
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Embodiment 1
[0039] Example 1: Isolation and identification of hydrogen-producing and anaerobic arsenic-methylating functional Clostridium sporogenes LHA6
[0040] The samples were collected from arsenic-contaminated paddy soil in Lianhua Mountain, Shantou City, Guangdong Province. Weigh 300g of the soil sample that has been naturally air-dried and sieved through a 10-mesh sieve into a 500ml anaerobic serum bottle, add 3g of straw powder and 300ml of sterilized ultrapure water, and aerate with high-purity nitrogen filtered through a 0.22μm filter head for 30 minutes. Serum bottles are sealed with sterile butyl rubber stoppers and aluminum ferrules. The soil anaerobic microorganisms were enriched by static culture in a 30°C incubator for 40 days. During this period, 200 μl of headspace gas products were collected with a gas sampling needle, and the headspace hydrogen concentration was detected by a high-performance gas chromatograph (Agilent 7820A, USA). 30 g of soil after pre-cultivation ...
Embodiment 2
[0051] Example 2: Arsenic methylation efficiency and hydrogen production effect of hydrogen production and anaerobic arsenic methylation function Clostridium sporogenes LHA6 at different methionine concentrations
[0052] In order to investigate the arsenic methylation and hydrogen production efficiency of LHA6 in anaerobic reactors containing different concentrations of methionine, LHA6 was expanded and cultured and then inoculated into different treatments. The details are as follows: Prepare 100ml of sterilized TSB liquid medium, inoculate the culture medium in an anaerobic glove box with a single colony after rolling the tube, seal it with a sterilized butyl rubber stopper and an aluminum ring, and culture at 30°C for 15 -20 hours, its OD600 is 0.4-0.5, which is in the middle of the logarithmic phase. Take 0.5ml of LHA6 bacteria solution and inoculate them in 8ml of TSB liquid medium with different treatments containing 30μM arsenous acid, the initial reaction bacteria OD6...
Embodiment 3
[0055] Example 3: Biomass growth assessment of hydrogen-producing and anaerobic arsenic-methylating functional Clostridium sporogenes LHA6 at different concentrations of arsenic and methionine
[0056] To evaluate the effect of different arsenic concentrations and methionine on the biomass of functional strain LHA6. With reference to the method of Example 2, on the basis of inoculating LHA6, arsenous acid (0, 30, 60, 100, 200 μM) and different concentrations of methionine (ie 0, 0.001, 0.01, 0.1, 1 , 10mM), cultured in a 15ml anaerobic reaction tube, and cultured statically in a 30°C constant temperature incubator for 7 days. The OD600 of the initial bacteria in the reaction was 0.1, and 1.0ml of the bacterial solution after 7 days of cultivation under different treatments was taken, and added to a light absorption standard microplate plate to determine the OD600 value.
[0057] Such as Figure 5 As shown, the OD600 of LHA6 thallus reached 0.8 after 7 days in the absence of ...
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