105 results about "Methane-Oxidizing Bacteria" patented technology

Methylotrophic or methanotrophic bacteria such as Methylobacterium are transformed with a gene of interest, and expression of the gene is regulated by means of a cumate repressor protein and an operator sequence which is operatively linked to the gene of interest, and the addition of an external agent. Specifically, the cymR repressor and cmt operator from Pseudomonas putida may serve to regulate gene expression in methylotrophic or methanotrophic bacteria with the addition of cumate.

A method of biosynthesis of polyhydroxyalkanoates (PHA) is provided that includes providing a type II methanotrophic bacteria, and disposing the type II methanotrophic bacteria in an unbalanced growth condition, where the unbalanced growth condition includes a nutrient-deficient media and a hydroxyalkanoic acid, and where the nutrient-deficient media has an absence of an essential nutrient required for cell replication of the type II methanotrophic bacteria.

The invention discloses a combined aerobic and anaerobic microorganism treatment system and method for gas in a mine goaf. The system comprises two parts: an aerobic methane-oxidizing bacteria agent spraying system and an anaerobic methane-oxidizing bacteria agent spraying system; the components comprise a methane-oxidizing bacteria agent storage tank, a pressure pump, a spraying pipe network, a nozzle and the like; the methane-oxidizing bacteria agent storage tank is connected with the input end of the pressure pump, and the output end of the pressure pump is connected with the spraying pipenetwork; the spraying pipe network is connected with the nozzle to form the spraying system. The method comprises the following steps: (1) culturing aerobic and anaerobic methane-oxidizing bacteria agents; (2) arranging the aerobic methane-oxidizing bacteria agent spraying pipe network system; (3) arranging the anaerobic methane-oxidizing bacteria agent spraying pipe network system; (4) regularlyspraying a methane-oxidizing bacteria agent and a culture solution; (5) testing effects. The danger of gas in the goaf can be reduced by regularly spraying the methane-oxidizing bacteria agent and theculture solution thereof into the goaf and removing methane by utilizing microbial metabolism.

The invention discloses a representation method of oil-gas exploration and oil-gas reservoir by taking vital bacterium abnormality and dead bacterium abnormality of methane-oxidizing bacteria as indicators, comprising the following steps: collecting a sample above an oil-gas exploration and oil-gas reservoir representation area according to the specific grid degree, the sampling depth and the sampling amount, packaging and saving the sample according to the specific condition; adopting a conventional microorganism counting method to obtain the vital bacterium quantity and the dead bacterium quantity of the methane-oxidizing bacteria in the sample; drawing an equal bacterium line of the vital bacteria and an equal bacterium line of the dead bacteria on a map, a topographic map, a geologic map or a topography-geologic map to obtain the vital bacterium abnormality and the dead bacterium abnormality; comprehensively comparing the vital bacterium abnormality and the dead bacterium abnormality; and estimating and predicting the underground oil-gas resource condition of the oil-gas exploration and oil-gas reservoir representation area, or extracting an exploitation scheme of the oil-gas reservoir according to the characteristics of the vital bacterium abnormality and the dead bacterium abnormality and the differences of the vital bacterium abnormality and the dead bacterium abnormality, and combining the results of geological exploration, geochemical exploration and geophysical exploration.

The invention discloses a preparation method of methane-oxidizing bacteria preparation and relates to a mixed culturing method of methane-oxidizing bacteria. Mineralized waste is employed as the carrier and the seed source of the methane-oxidizing bacteria. Enrichment of the methane-oxidizing bacteria is carried out on the mineralized waste; then the enriched mineralized waste is added into the culture fluid of the improved formula of NMS culture fluid used for culturing normal methane-oxidizing bacteria; then the mixed gas of methane and oxygen with a volume ratio of 1:1 to 1: 1.5 is pumped to carry out culture. When the method is employed, the effect for culturing the methane-oxidizing bacteria is good and the starting is so quick that the largest methane oxidation rate can appear in about 2 days. The mixedly cultured methane-oxidizing bacteria avoid the problems that the traditional strain culture method and axenic isolation process are complex and the pure strains are not easily preserved. The preparation of the invention has a simple preparation method, and a remarkable methane oxidizing effect; moreover, the bacteria liquid is easy to be stored and transported. The preparation can be conveniently used for controlling the methane discharge in a refuse landfill and can also provide the methane-oxidizing bacteria strain for industrial production.

The invention relates to a membrane biological treatment device and method for cooperative treatment of methane and sulfur and nitrogen-containing wastewater. The device provided by the invention comprises a gas membrane separation container provided inside with a plurality of gas membrane modules, a circulation type oxidation container for holding a filling material adhered with methane-oxidizing bacteria and sulfate reducing bacteria, and a suspension, an upflow-type regeneration container for holding a supernatant solution containing Thiobacillus denitrificans, a solid-liquid separation container and a gas guiding container. Methane is oxidized and sulfate is reduced in the circulation type oxidation container; separation effect of gas membrane modules make unoxidized methane return to the circulation type oxidation container for continuous oxidization, and make the sulfate reducing product hydrogen sulfide enter into the upflow-type regeneration container and be oxidized by nitrate into sulfate, as a regeneration oxidant; and at the same time, nitrate is reduced into nitrogen, so as to realize cooperative effective treatment of methane exhaust gas and sulfur and nitrogen-containing wastewater in two biochemical reaction zones. The device provided by the invention has the advanategs of compact structure, low energy consumption and simple operation.

The invention relates to a sewage treatment system and a sewage treatment method for reducing greenhouse gas emission. The system comprises a moving bed biomembrane biochemical reactor MBBR, an air stripping pool and an up-flow anaerobic sludge blanket UASB which are sequentially connected, wherein bio-filler is arranged in the MBBR, an aerating head is arranged at the bottom, an intercepting screen is arranged at an upper overflowing water outlet for preventing loss of the bio-filler, a mud outlet is formed at the bottom of the UASB, a sludge return opening is formed on the upper part, and a three-phase separator and an air outlet are formed on the top. Sewage enters into the MBBR for intermittent aeration, so that accumulation of NO<2-> is realized; effluent of the MBBR enters into the air stripping pool through a peristaltic pump to remove dissolved gas in water and then enters into the UASB; denitrifying anaerobic methane-oxidizing bacteria convert NO<2-> into N2 to be removed. According to the system provided by the invention, the UASB is coupled by using the MBBR, so that actual sewage with low carbon nitrogen ratio is treated in an energy-saving and efficient manner with less secondary pollution.

The invention discloses a preparation method for methane-oxidizing bacteria liquid. The preparation method comprises the following steps that mineralized refuse is mixed with leachate, cultivation is conducted on the condition that mixed gas of methane and oxygen is input, the methane-oxidizing bacteria liquid is obtained, and the methane oxidization rate of the mineralized refuse exceeds 50 %. The invention provides refuse landfill covering materials and a refuse landfill methane emission reduction method. The mineralized refuse is used as the source of the methane oxidation bacteria strain, the leachate is used as a culture medium to culture the methane oxidation bacteria in a mixed mode, the cultivation system is close to the environment of the landfill, oxidization efficiency of the methane in landfill gas is higher, and the methane oxidation rate can exceed 84 % when covering is conducted for 7 days.

The invention discloses a methane gas thin-layer medium electrochemical biosensor and a preparing method thereof. The biosensor comprises a counter electrode, a section at one end of the counter electrode is reserved for being connected with an electrochemical workstation when methane gas is detected, the remaining part of the counter electrode is wrapped with electrolytic paper, the outer portion of the electrolytic paper is wound by a carbon fiber monofilament, one end of the carbon fiber monofilament is suspended and used for being connected with the electrochemical workstation when methane gas is detected, and methane-oxidizing bacteria are attached to the carbon fiber monofilament. The concentration of methane gas is detected through an electrochemical method, the high-temperature condition of a heat conduction method is not needed, and safety is fully ensured under the normal-temperature solution condition. The methane-oxidizing bacteria serve as a biological catalyst, and the detection selectivity and sensitivity on methane gas are improved through the features of high specificity and high selectivity of the biological catalyzing process.

The invention relates to the field of garbage treatment, in particular to a final covering layer for household refuse semi-aerobic landfill. The final covering layer covers the surface layer of a household refuse semi-aerobic landfill and comprises a vegetation layer, a biological covering layer, a reservoir layer, a capillary barrier layer and a drainage layer sequentially from top to bottom. Methane-oxidizing bacteria exist in the biological covering layer. According to the actual situation of the household refuse semi-aerobic landfill process, the covering layer provided by the invention has good air permeability, and it can be guaranteed that external air is diffused into a landfill pile body; and meanwhile, the landfill gases including methane and the like generated when pile body landfill is conducted are diffused outside the pile body from the inside, so that an appropriate oxidation removing environment is provided for the methane released into the air through the covering layer. By adoption of the final covering layer for household refuse semi-aerobic landfill, the methane oxidation efficiency of the final covering layer is improved through optimization design of all the layers.

The present invention discloses process and special bacterial colony for degrading organic phenol matter. Mixed bacterial colony, MY9 CGMCC No. 1893, containing methylosinus trichosporium is inoculated to organic phenol matter liquid to be degraded, and cultured in sealed mixed gas environment containing methane and air. The process of degrading organic phenol matter utilizes the synergistic metabolic action of methane oxidizing bacteria and other microbes in the said mixed bacterial colony in degrading organic phenol pollutant, and has mild reaction condition and high efficiency.

The invention discloses a compost preparation method for reducing emission of greenhouse gases in kitchen garbage compost. The compost preparation method comprises the following steps: mixing matured compost, kitchen garbage and maize straws, thereby obtaining mixed compost material with the mass moisture content of 50-60 percent; performing aerobic fermentation on the mixed compost material, thus obtaining the compost, wherein the matured compost is obtained by composting and decomposing wet-based kitchen garbage and maize straws of which the wet bases are in a mass ratio of 17:3-9:1; and a mass ratio of the wet base of the matured compost to the wet base of the mixture of kitchen garbage and maize straws is 1:9-1:4. Compared with the prior art, the method disclosed by the invention has the advantages that (1) the recycled matured compost is soft in texture and large in specific surface area, has high adsorption capacity and is enriched in lots of methane-oxidizing bacteria and nitrite oxidizing bacteria, and the emission of greenhouse gases such as CH4 and N2O in the composting process can be obviously reduced; and (2) the method disclosed by the invention is easy to operate and low in cost and has significant application values in emission reduction and regulation of pollution gases in the kitchen garbage composting process.

The invention disclosure is a method of bioremediation of wastewater, particularly groundwater, by utilizing coupled anaerobic and aerobic biological treatment, more specifically, methanogenic (strictly anaerobic) and methanotrophic (strictly aerobic) microbial populations, in combination with a supply of in-situ generated water-dissolved oxygen and hydrogen. Water electrolysis is used to produce water-dissolved oxygen and hydrogen. The immediate advantage of using H2 from the electrolysis is to provide electron donors to methanogens to reductively dechlorinate the chloroaliphatics, and to reduce the water carbonates and generate methane which is used as energy and carbon source for the methanotrophic bacteria. Oxygen is used as electron acceptor by the aerobic bacteria, including the methanotrophs. The addition of an organic carbon source can be minimized or even eliminated, so as to diminish the competition between methanotrophic bacteria and heterotrophic bacteria for oxygen.

The invention discloses an oil-gas exploration method based on anomaly of light hydrocarbon oxidizing bacteria, which comprises the following steps: after homogenizing a soil sample, extracting sample DNA (deoxyribonucleic acid), and quantifying propane oxidizing bacteria and methane oxidizing bacteria by a fluorescence quantitative PCR (polymerase chain reaction) technique; determining anomaly thresholds of the propane oxidizing bacteria and methane oxidizing bacteria, establishing a microbiological anomaly reference system for verification zone oil-gas microbiological exploration, drawing isoline of the propane oxidizing bacteria, and carrying out microbiological anomaly zoning; and marking the gas-cap reservoir and pure reservoir in the anomaly zone according to the content of the methane oxidizing bacteria. According to the method, the anomaly of light hydrocarbon oxidizing bacteria is utilized to perform oil-gas exploration; in the precision detection aspect, the quantity of the light hydrocarbon oxidizing bacteria in the sample is directly detected to avoid the artificial interference in the traditional culture method; and the detection quantity is higher than that of the traditional culture method by 1-2 orders of magnitude, the time is shortened to 12 hours from 14-21 days in the traditional culture method, and the method can detect 96 samples at one time, thereby effectively enhancing the precision and detection efficiency of microbial oil-gas exploration.

The invention discloses an effective microbiological detection method capable of being applied to oil-gas exploration, belongs to the field of biotechnology, and relates to a method for detecting unusual methane-oxidizing bacteria content, which can be applied to microbial oil-gas exploration, takes methane-oxidizing bacteria as indicators and can effectively avoid the influence of biomethanation. The methane-oxidizing bacteria and methanogens in a soil sample are respectively quantified by a Real-time polymerase chain reaction (PCR) technology, the methane-oxidizing bacteria content and ratios of the methane-oxidizing bacteria to the methanogens in different regions are compared, and that whether the reason of unusual methane-oxidizing bacteria is biomethanation is further deduced, so that the influence of the biomethanation is eliminated and risks of false negative results are reduced. The method can be applied to the microbial oil-gas exploration, provides cheap and effective help for the early stage of the exploration, and indicates and predicts favorable exploration blocks to reduce exploration risks and cost.

The invention discloses a multilayer vertical methane biological oxidation device, relating to the technical field of greenhouse gasses abatement, wherein the device is composed of a reactor, methane supply system and air supply system. Methane and air is forcibly led to flow along the pores of filling medium up to down in the device by the gas supply system. Plurality of gas-arranging layers are vertically mounted in the reactor to realize the frequent mixing for the gas to reduce the short flow of the gas, provide growth advantage for methane oxidation bacteria in the filling medium, and make the methane to be continuously decomposed in the device to transform into CO2 and discharge from the outlet at last.

The invention discloses a method for removing methane in the environment, comprising the following steps: 1) collecting soil sample of a coal mine or a refuse landfill; 2) adding the soil sample of the step 1) into an NMS culture medium, and culturing in a sealed bottle; when in initial period, leading the volume ratio among the NMS culture medium, the methane and air in the sealed bottle to be (5-30): (5-40): (30-80); replacing the methane into the sealed bottle every 24-72h, leading the volume ratio among the NMS culture medium, the methane and air in the sealed bottle to be (5-30): (5-40):(30-80); 3) inoculating the culture solution of the step 2) into a new NMS culture medium for passaging every 8-12 days; leading the volume ratio between the culture solution of the step 2) and the NMS culture medium to be (5-15): (95-85); carrying out passage for more than ten generations, and obtaining mixed methane-oxidizing bacteria; and 4) culturing the mixed methane-oxidizing bacteria obtained from the step 3), and removing the methane in the environment. The method for removing the methane in the environment has simple operation, strong methane removing capability and wide application scope, and has broad application prospect for removing coal-mine gas, reducing emission of landfill gas of the refuse landfill and the like.

The present invention discloses one kind of recombinant bacterium with granular methane monooxygenase activity and its application. Fusion gene comprising the promoter sequences of coding gene and desulfurizing gene of granular methane monooxygenase is introduced into Rhodococcus, and after screening, the strain expressing granular methane monooxygenase or the recombinant bacterium with granular methane monooxygenase activity is obtained. The promoter sequence of the desulfurizing gene is located in the upstream of the granular methane monooxygenase coding gene. The recombinant bacterium avoids the effect of methanol dehydrogenase radically, makes methane oxidation stop in methanol reaction step, and is superior to wild methane oxidizing bacteria and easy to culture. The recombinant bacterium can convert alkane in natural gas into liquid alcohol, convert methane into methanol and oxidize propylene into 1, 2-epoxy propane.

The invention discloses a method for culturing a methane-oxidizing bacterium, which adds cetane in the culture medium based on the conventional method of the methane-oxidizing bacterium. Cetane is sheared by the mechanical force so as to form the micro dispersion droplet in the process that methane-oxidizing bacterium is shake cultured, the droplets can improves the dissolvability of methane and the cell can shuttle between the water phase and the organic phase owing to the compatibility of the bacterial cell. The invention can improve the growth speed and the cell density of the methane-oxidizing bacterium, which has the simple culture method, and has the wide application prospect.

The invention publishes a kind of biology method to prepare methanol using methane catalysis. The method adopts the technology as follows. Fist, bacterial cells are oxidized by methane and the outcome is as catalyst. Second, add mixed gas of methane, carbon dioxide and oxygen to membrane reactor and start up reaction under the condition of 32-40deg.C and 0-1.2MPa. after the reaction the aqueous solution containing methanol can be gotten. The invention solves the problems that cytoactive will depress because of the consumption of NADH lead by chemical inhibitor. In this method, the reaction can continue to make sure that coenzymes NADH can rebirth in situ in the reaction system.

The invention relates to the technical field of sewage treatment, and concretely relates to a method for synchronously realizing sewage denitrification and biogas desulfurization. The method comprisesthe following processes: internal carbon source removal treatment, preliminary denitrification treatment, and sewage denitrification and biogas desulfurization synchronous decoupling treatment. In the process of the internal carbon source removal treatment, a nitrate acting as an electron acceptor can metabolize the self carbon source of sludge; in the process of preliminary denitrification treatment, methane used as an electron donor is used for domesticating and cultivating methane-denitrifying bacteria to achieve anaerobic methane oxidation-based nitrate denitrification; and in the processof sewage denitrification and biogas desulfurization simultaneous coupling treatment, hydrogen sulfide and methane in biogas, which act as a common electron donor enrich autotrophic denitrifying bacteria in the presence of anaerobic methane-oxidizing bacteria, and provide a good environment for the co-growth of the autotrophic denitrifying bacteria and anaerobic methane-oxidizing bacteria in order to realize the coupling of the two processes of sewage denitrification and biogas desulfurization.

The invention provides a method of electrically-assisting electroactive methane oxidizing bacteria to catalyze methane to produce methanol or formic acid. According to the method provided by the invention, the dehydrogenase activity of the electroactive methane oxidizing bacteria is controlled by a redox potential which is controlled by a microorganism electrolysis pool, and the activity and the stability of the electroactive methane oxidizing bacteria are maintained while methanol or formic acid, serving as a chemical product, is obtained; H<+> and electrons are produced while the electroactive methane oxidizing bacteria which are adhered to the surface of an anode of the microorganism electrolysis pool metabolize methane to producemethanol or formic acid, and H<+> and electrons are combined on the surface of the cathode under the action of electrically assisting and a cathode catalyst. The method is capable of solving the problems of synthesizing methanol or formic acid by traditional chemical reaction of high energy consumption and low efficiency of the cathode catalyst and preventing further deep oxidation of methanol and formic acid to produce CO2 in the process of bio-catalytically oxidizing methane to produce methanol or formic acid; according to the method provided by the invention, the whole reaction process is carried out under mild conditions and is environmentally friendly.

The invention discloses a culture device and a culture method for double membrane aerating aerobiotic methane-oxidizing bacteria. The method comprises the following steps: putting sludge rich in methane-oxidizing bacteria and a soil suspension into a reaction device cavity; and introducing methanol and oxygen by way of bubble-less aeration to achieve high-density and low-cost culture of the aerobiotic methane-oxidizing bacteria. The invention also provides the culture device for double membrane aerating aerobiotic methane-oxidizing bacteria. As solubility of methane and oxygen in water is relatively low, sufficient carbon sources and energy sources cannot be provided to microorganisms in a reaction device and methane mixed in air or oxygen exceeding a certain proportion has a risk of explosion, gas-liquid mass transfer resistance is broken through by way of double membrane aeration, so that the dissolved load of methane and oxygen in a solution is improved. A primer can be provided efficiently at low cost, and the culture device and culture method are suitable for culturing aerobiotic methane-oxidizing bacteria efficiently, and a high density aerobiotic methane-oxidizing bacteria culture is used for sewage treatment, intermediate product production, greenhouse gas emission reduction and the like.

The invention discloses a synchronous methanogenesis and aerobic methane oxidation coupling denitrification process control method. The method comprises the following steps: (1) establishing a reaction system; (2) carrying out microbial culture; (3) enriching anaerobic methanogens; (4) enriching aerobic methane oxidizing bacteria; and (5) enriching denitrifying bacteria, removing 91.4%-98.8% of COD, 93.2%-99% of NO3-N and 71%-93.5% of TN by a system under the process condition, and ensuring that the methane recovery amount is 180-660mL / d. The method is economical and efficient, and good socialand economic benefits can be generated. The organic and nitrogen pollutants in the wastewater are removed, the greenhouse effect of effluent soluble methane is eliminated, and meanwhile part of gaseous methane can be recycled.

The invention belongs to the technical field of biological analysis, and in particular provides a method for simultaneous analysis of ammoxidation or methane oxidation correlated bacterial community structure. The method comprises the following steps: designing a high-degeneracy tag primer capable of covering a particulate methane oxidase alpha subunit gene and an ammoxidation monooxygenase alpha subunit gene; conducting PCR amplification on a genomic DNA extracted from an environmental sample by using the primer; conducting PCR amplification on the purified PCR products from the previous step by using a barcode marked tag primer; sequencing the amplified sequences; and analyzing to obtain ammoxidation or methane oxidation correlated bacterial community structure characteristics. The fidelity of the method is verified by different pmoA / amoA gene recombinant plasmid mixture experiments, so the method can significantly improve PCR amplification effect on the premise of improving primers coverage degree, and truly reflect the distribution of the different communities in environmental sample. The method completes structure analysis of ammonia oxidation and methane oxidation bacterial community in the environmental sample, and is applicable to structure analysis of other functional microflora.

The invention publishes a kind of biology method to prepare methanol using carbon dioxide catalysis. Fist, use methane to oxidize Methylosimus trichosporium IMV as the catalyst. Second, add carbon dioxide or the mixture of carbon dioxide and air as reaction substrate. In this method, the outcome methanol can be gotten and the transformation from carbon dioxide to methanol.

The invention relates to a microbacterium capable of utilizing methane and the applications thereof. The CCTCC NO of the microbacterium is M2010099. The microbacterium can be used to prepare the methanogen absorbent, the garbage treatment agent and the biocatalyst. The microbacterium overcomes the application problems that the methane oxidizing bacteria have low microbial density, the cultivation of the bacteria is difficult to enlarged, etc; and the microbacterium has higher affinity to methane. The microbacterium is especially suitable for man-made methane sources such as domestic garbage landfills and rice fields and can be widely popularized.

The invention relates to a method for preparing methane-oxidizing bacteria on a large scale. The method includes the following steps: 1), putting the methane-oxidizing bacteria and inorganic salt into a reactor, and then injecting methane and oxygen into the reactor; 2), crushing the added gases into tiny bubbles by a crushing device in the reactor to enable the bubbles to be blended in liquid in the reactor and flow along the reactor internally together with the liquid, and preforming a fermentation process; 3), after the fermentation process is finished, determining completion of the fermentation process, performing initial condensation of a product in the reactor, and quickly cooling and drying the product after heating to obtain an initial product. The method has the advantages that the method mainly aims at preparation for methane-oxidizing bacteria production of large-scale reaction devices, the methane-oxidizing bacteria are cultivated by the devices such as the reactor, water solubility of the gases such as the methane is improved through measures such as bubble crushing, reaction efficiency is improved through addition of inorganic salt cultivation media, and accordingly, the method can be suitable for large-scale production application.

The present disclosure relates to genetically engineered methanotrophic bacteria with the capability of growing on a multi-carbon substrate as a primary or sole carbon source and methods for growing methanotrophic bacteria on a multi-carbon substrate.

The invention discloses a membrane biological reaction system for treating greenhouse gas by bacteria and algae synergistically. The system comprises a reactor, a bio-membrane system and an illumination device, wherein the bio-membrane system is arranged in the reactor and divides the internal space of the reactor into a methane-oxidizing bacterium reaction chamber and a microalgae reaction chamber; the bio-membrane system comprises a microalgae bio-membrane and a methane-oxidizing bacterium bio-membrane; a microporous filter membrane is arranged between the microalgae bio-membrane and the methane-oxidizing bacterium bio-membrane; and the illumination device is arranged right in front of the microalgae reaction chamber. In the membrane biological reaction system, oxygen generated by microalgae photosynthetic carbon sequestration and carbon dioxide generated by biochemical conversion of methane-oxidizing bacteria can be utilized by the methane-oxidizing bacteria and the microalgae respectively, bacteria and algae metabolic processes are coupled, a conversion process is clean, the product is supplied to cell growth and metabolism, adsorption type culture is energy-saving and efficient, and the purposes of synergistically treating greenhouse gas and increasing the biomass yield can be achieved.