45 results about "Resazurin" patented technology

The invention discloses a novel clostridium beijerinckii strain for producing butanol. The strain has a classification name of clostridium beijerinckii IT66 (Clostridium beijerinckii IT66) and has a collection number of CCTCCM NO:2011404. The invention also discloses application of the clostridium beijerinckii in fermentation production of the butanol. The clostridium beijerinckii IB4 is subjected to continuous culture and plasma induced mutation by adopting a chemostat, and a strain which tolerates an inhibitor and is high in reducing power is screened through a resazurin flat plate and is high in acidolysis byproduct resistance, high in butanol ratio and high in solvent yield. When a non-detoxified corncob acidolysis sugar solution is a carbon source, the total solvent yield and butanol yield in a 5L fermentation tank respectively reach 11.3g/L and 9.1g/L and are respectively improved by 2.3 times and 2.4 times compared with those of an original strain cultured under the same conditions, the butanol ratio is 80 percent, the sugar conversion rate is 0.32, and the strain has significant social meaning and economic value.

The invention discloses a separated purification method and the application thereof of a polycyclic aromatic hydrocarbon anaerobic degrading pure strain, which belongs to the polluted soil and aqueous biological degradation handling field. A base bottom layer flat plate is manufactured in the a anaerobic box filled with nitrogen gas by taking polycyclic aromatic hydrocarbon as a sole carbon source and an energy source and taking sodium nitrate or potassium nitrate as an electronic accepted anaerobic agar solid culture, the top layer flat plate of the anaerobic agar solid culture including polycyclic aromatic hydrocarbon anaerobic degrading mixed bacteria is manufactured on the bottom layer flat plate, the manufactured culture medium flat plate is put in a drying container filled with the nitrogen gas to culture one to two months after being sealed. Polycyclic aromatic hydrocarbon, electronic acceptance, basic culture medium, trace metal liquid, vitamin c solution and colony in the culture medium flat plate connected into the anaerobic bottle of resazurin are added in the anaerobic box filled with the nitrogen gas, and are cultured for 20 to 25 days in an oscillator under the temperature of 20 plus or minus 2 DEG C and the rotating speed of 100 r / min. Then, the cyclical switchover is performed, after a cycle time is larger than four times, the pure strain of the anaerobic degrading polycyclic aromatic hydrocarbon can be obtained. The method can obtain good microbe pure strain to the polycyclic aromatic hydrocarbon anaerobic degrading performance by being separated.

The invention discloses a method for rapidly screening and evaluating an antibiotic bioactivator, in particular a test method for screening the antibiotic bioactivator by combining an accelerated solvent extraction apparatus and a microplate resazurin assay. The method comprises the following steps of: preparing a bioactivator by the accelerated solvent extraction apparatus; adding the bioactivator and known antibiotic into a microplate according to concentration gradient, and adding experimental bacterial suspension into various holes; placing the microplate under suitable conditions, incubating, adding resazurin solution, and mixing uniformly; determining light absorption values of the holes of the microplate at 600nm by using a microplate reader; evaluating a minimum inhibitory concentration (MIC) value of the bioactivator to be screened according to light absorption values in known antibiotic holes and light absorption values in bioactivator holes at different concentrations; and further evaluating the antibiotic activity of the bioactivator. The method is rapid, simple and convenient, and the antibiotic activity of the bioactivator can be objectively and accurately evaluated, so that the high-throughput screening of the antibiotic bioactivator becomes possible.

The invention discloses a method for producing hydrogen through intensified anaerobic fermentation. The method comprises the following steps of: adding a culture medium, resazurin and a cetyl trimethyl ammonium bromide solution into a fermentation container, mixing uniformly, sterilizing, inoculating liquid obtained after cow dung is fermented, introducing sterile nitrogen into the fermentation container and an alkaline washing bottle which is connected with the fermentation container so as to displace the air, sealing a sterile nitrogen introduction port, performing shake culture at constant temperature, performing alkaline washing on gas produced by fermentation, and collecting to obtain the hydrogen. By adding cetyl trimethyl ammonium bromide into a fermentation substrate, the problems that the cost of the current hydrogen production by biological anaerobic fermentation is still high and the hydrogen production efficiency is low are solved, and the effects that the accumulated hydrogen yield is improved by 38 percent and the maximum hydrogen production rate is improved by 44 percent are successfully achieved. The method can be simply operated, and is low in cost and easy to implement.

The invention provides a method for producing hydrogen by fermenting special anaerobic clostridium butyricum. The screened special anaerobic bacterium is clostridium butyricum JCM1391. Proven by tests, the hydrogen is produced by fermenting by batch by using anaerobic bacteria under the sterile anaerobic condition, wherein the mixture of 10.0-15.0g of peptone, 2.0-3.0g of beef extract, 5.0-10.0g of yeast extract, 3.0-4.0g of Na2HPO4, 0.2-0.5g of L-cysteine, 0.2-0.5g of L-cysteine hydrochloride monohydrate, 1 ml of resazurin indicator which accounts for 0.2 percent by weight and 1 ml of distilled water is used as culture media, and glucose is used as fermentation matrix. The method has the advantage of high hydrogen production efficiency and low requirement on environmental condition for fermentation.

The invention relates to a Clostridium acetobutylicum strain capable of quickly generating butanol by fermenting xylose and starch, and a screening method and application thereof. The invention discloses Clostridium acetobutylicum, which is classified and named Clostridium acetobutylicumXY16 with the preservation and registration number of CCTCC NO:M2010011. The strain obtained by ion beam mutation and screening of a xylose plate, a 2-deoxy-D-glucose plate, a bromocresol green plate and a resazurin plate can quickly and efficiently generate the butanol by fermenting xylose and starch, has the advantages of high yield of total solvent, high butanol ratio and good repeatability, and is a good strain suitable for industrial production.

The invention provides a bergenia crassifolia extract and an extracting method and medical application in oral diseases. Through a resazurin dyeing method and an XTT method, the extract of bergenia crassifolia is subjected to bacteriostatic activity and inhibitory biological film activity study, the extract of bergenia crassifolia has an obvious inhibitory effect on streptococcus mutans, streptococcus sanguis, streptococcus salivarius, lactobacillus acidophilus, porphyromonas gingivalis, fusobacterium nucleatum, actinobacillus actinomycetemcomitan and biological film of the fungi, has a better inhibitory effect on streptococcus mutans, porphyromonas gingivalis and the biological film of the fungi and can be used for preparing daily necessities and medicine for preventing and treating the oral diseases.

The invention discloses a method for screening or detecting antibacterial active substance from plant extract. The method includes the following steps: (1) plant extract is prepared, or an extract bank is created based on the plant extract, or constituent or ingredient is obtained through separation and purification; (2) cultivation is completed according to the culture conditions of different bacteria and fungi; (3) each sample to be tested is added in bacterium culture solution or fungus culture solution; (4) 12 hours to 24 hours cultivation is completed under bacterium culture condition or fungus culture condition; (5) resazurin indicator is added in each culture solution; (6) according to the color change of resazurin, bacterium survival status is judged so as to judge whether the constituent or ingredient of the plant extract has bacteriostatic activity. With easy operation, simple experiment apparatus and less work load, the invention can obtain obvious results; moreover, the invention realizes fast screening of plant extract with antibacterial activity and screens out antibacterial active constituent or ingredient from the plant extract quickly, thereby providing basis for drug research; in addition, the invention can also be used in detecting plant antibacterial active constituent.

The invention discloses a method for detecting D-2-hydroxyglutarate by using FAD dependent D-2-hydroxyglutarate dehydrogenase and resazurin, and relates to the technical field of enzymatic detection.The method uses FAD dependent D-2-hydroxyglutarate dehydrogenase, resazurin and a PBS buffer solution to realize detection of D-2-hydroxyglutarate; and the FAD dependent D-2-hydroxyglutarate dehydrogenase is an AX-F-D2HGDH protein or a P-D2HGDH protein. The method is simpler in composition, lower in cost and simpler and more convenient to operate. Since no cofactor, coenzyme or diaphorase needs tobe added, introduced interference is less, sensitivity is higher, only one enzyme needs to be prepared, no exogenous cofactor NAD and the like need to be added, and meanwhile, the buffer solution canbe a PBS buffer solution which is lower in cost and easy to obtain, so that cost is lower.

The invention relates to a display experiment method and device for coupling bubble internal flow and bubble external mass transfer. The method comprises the following steps of adding a prepared resazurin solution with a certain concentration, a sodium hydroxide solution and a glucose solution to a certain liquid level in a bubble observation chamber by utilizing a resazurin chromogenic reaction and taking smoke as tracer particles, introducing a mixed gas of oxygen and smoke, and by virtue of a high-speed camera and an image processing system, meanwhile, the phenomena of fluid flowing inside the bubbles and liquid phase mass transfer outside the bubbles being displayed. The method is advantaged in that an internal flow phenomenon of the captured bubbles is obvious, and the mass transfer wake outside the bubbles is clear.

The invention discloses a pure culture method of anaerobic microorganisms. The pure culture method comprises the following steps of: step 1, preparing a liquid culture solution and boiling; dropwise adding resazurin; sub-packaging into a thin-opening pipe to obtain a culture pipe; step 2, introducing N2/H2 mixed gas and aerating; rapidly covering a rubber plug tightly; step 3, putting the culture pipe into a high-pressure sterilization pot and sterilizing at high temperature; transferring into a super-clean platform and carrying out ultraviolet sterilization; and step 4, injecting a magnetic antioxidant into the culture pipe by utilizing an injector and uniformly mixing; injecting an anaerobic microorganism seed solution into the culture pipe by utilizing the injector; putting the culture pipe into a constant-temperature culture box and culturing to obtain an anaerobic microorganism group. The pure culture method of the anaerobic microorganisms has the beneficial effects that (1) an anaerobic culture box does not need to be utilized and the operation is simple and convenient; and (2) the magnetic antioxidant is not dissolved into the liquid culture solution and is not easily absorbed by the anaerobic microorganisms, so that the growth is not influenced; the separation of the anaerobic microorganisms can be simplified through magnetic separation and the relatively pure anaerobic microorganism group is obtained.

The invention discloses a method for rapidly identifying anaerobic microorganisms in wine brewing, and belongs to the technical field of anaerobic microorganism identification. The method for rapidlyidentifying the anaerobic microorganisms in the wine brewing comprises the following steps: S1, preparing an anaerobic culture solution, namely uniformly mixing 0.18 g of potassium persulfate, 0.2993g of potassium phosphate, 0.35 g of sodium chloride, 0.40 g of ammonium persulphate, 0.040 g of magnesium chloride, 0.085 g of potassium sulfate, 0.002 g of resazurin, 0.6 g of L-cysteine, 0.7 g of L-ascorbic acid, 5.0 g of sodium bicarbonate, 1.3 g of peptone and 1500 mL of distilled water so as to obtain an anaerobic culture solution for standby application; S2, preparing a bacterial suspension,namely weighing 15 g of fermented grains in the presence of a nitrogen flow, adding the weighed fermented grains into 95 mL of sterilized normal saline in an anaerobic bottle, quickly and tightly covering the anaerobic bottle with a cap, oscillating the anaerobic bottle on a shaking table at a temperature of 50 DEG C and a rate of 200 r/min for 15 min, taking the oscillated anaerobic bottle out,putting the anaerobic bottle into an anaerobic workstation, sucking 0.2 mL of liquid supernatant, and adding the liquid supernatant into the sterilized culture solution so as to carry out anaerobic accumulation culture for 36 hours; S3, carrying out colony culture; and S5, carrying out identification. The method for rapidly identifying the anaerobic microorganisms in the wine brewing is convenientto operate, simple in processm, and hign in identification accuracy.

The invention discloses an intestinal anaerobic microorganism culture bottle and a preparation method thereof. The culture bottle comprises a culture bottle sealed and packaged and a culture solutionfilled in the culture bottle. The prepration method comprises the steps of sequentially mixing a basic culture medium solution, an L-cysteine saline solution and a resazurin aqueous solution in proportion to obtain a basic culture solution, filling the culture bottle with the basic culture solution, and sealing and packaging the culture bottle; performing high-pressure sterilization on the sealedculture bottle; and finally, mixing a 1640 culture medium solution, BI serum, vitamin K and hemin in proportion to obtain an active additive mixed solution, injecting the active additive mixed solution into the culture bottle which is subjected to high-pressure sterilization and cooled to room temperature, thereby completing the preparation. According to the anaerobic microorganism culture bottle,a culture bottle body and a packaging piece are adopted, all components for preparing the culture solution are commercially available products, the raw materials are easy to obtain, the threshold andcost of anaerobic culture are reduced, anaerobic culture, storage and transportation are easier to operate, the preparation method is simple and easy to implement, the requirement for large-scale production is met, and the use requirements of hospitals and scientific research institutions are met.

The invention belongs to the technical field of microbial engineering. In order to solve the problems that the existing strain preservation method cannot meet the requirements of short-term and long-term preservation of strains of methanogens produced by coal-bed methane and cannot maintain the activity of high-yield methane gas of the strains, the invention provides a strain preservation method of anaerobic methanogens in coal seams. The method comprises the steps of screening a strain to be preserved, adding the screened strain into a 5ml of anaerobic tube, then adding an equivoluminal sterilized glycerin/deoxidized L-cysteine solution, evenly mixing the mixed solution, and then preserving; long-term preservation: mixing lump anthracite coal, a preservation nutrient solution, L-cysteine, resazurin and a bacterium solution according to a certain ratio and then preserving the mixture at room temperature, simulating the environmental conditions of the coal seams, and enabling the methanogens to maintain higher activity and a more stable bacterial flora structure all the time so as to obtain more ideal methane yield. The strain preservation method does not need ultra-low temperature equipment and the like, does not need special equipment, is convenient and fast, is suitable for short-term and long-term preservation of different amounts of strains, is economical and convenient, and lowers the production and test costs.

The invention discloses a method for detecting PBMC activity under an in-vitro 3D culture condition by using a resazurin reagent. The method comprises the following specific steps: step 1, preparing resazurin mother liquor; step 2, preparing a resazurin detection reagent; and step 3, determining the activity of PBMC under an in-vitro 3D culture condition. The standard resazurin solution which canbe produced on a large scale and is used for detecting the activity of human peripheral blood mononuclear cells after being cultured in an in-vitro 3D culture system in vitro is established; the reagent is high in cell activity detection sensitivity, convenient to operate, free of adverse effects on cells and free of pollution to the environment, and it is proved through actual tests that the reagent is suitable for detecting the activity of PBMC in a 3D culture system.

The invention relates to a method for quickly screening anti-Aspergillus-niger active substances, which comprises the following steps: adding a sample to be screened, a resazurin and Aspergillus niger spore bacterial suspension, into a 96 microporous plate, and culturing in a 37 DEG C constant-temperature culture box for some time to obtain the result, wherein a negative control and a positive control are arranged, and the preferable culture time is 12-36 hours; and screening to obtain the anti-Aspergillus-niger active substances according to the result. When the color of the microporous plate is blue or purple, the fluorescent value measured by the microplate reader is lower than the 15000; and when the inhibition rate is higher than 40%, the detected sample is the anti-Aspergillus-niger active substances. The method is simple, quick and accurate.

The invention provides a deoxidizing method of a fluid medium. The method comprises the following steps: S1, preparing a resazurin water solution to serve as an anaerobic indicator which takes on a dark blue color or a light blue color in an oxygen dissolution state, takes on a pink color in a slight oxygen dissolution state, and is colorless in an oxygen dissolution-free state; S2, adding L-cysteine hydrochloride, a biological buffering agent MES (morpholineethanesulfonic acid) and an anaerobic indicator into an anaerobic bottle with 1L of the fluid medium and mixing uniformly; S3, introducing nitrogen gas to the bottom of the anaerobic bottle in the S2 for 50-100min, pretreating a part of dissolved oxygen in the fluid medium to change the color of the fluid medium from the dark blue color to the light blue color or the light pink color; and S4, enabling the fluid medium of which a part of the dissolved oxygen is removed to stand, and consuming the rest of dissolved oxygen in the anaerobic bottle through the added L-cysteine hydrochloride in the S2 to change the color of the fluid medium from the light blue color or the light pink color to an achromatic color, thus completing deoxygenization. The deoxidizing method can be used for removing the dissolved oxygen in the fluid medium, and is conducive to enrichment and cultivation research of anaerobic micro-organisms.

The invention discloses a method of screening a 1-deoxyxylulose-5-phosphate racemase inhibitor from a marine microorganism fermentation broth. The method includes: 1) a step of preparing a marine microorganism fermentation broth sample, namely, a step of inoculating a fungus, fermenting, extracting metabolites, concentrating and eluting; 2) a step of designing a primer and performing PCR amplification by adopting the whole genome of vibrio vulnificus as a template, cloning a vector, converting escherichia coli, inducing expression of a target protein, and subjecting the target protein to elution, dialysis, ultrafiltration and desalting to obtain His-DXR; 3) a step of reacting a bacterial suspension, a resazurin solution and the marine microorganism fermentation broth sample obtained by the step 1), measuring the fluorescence value, calculating the inhibition rate through an equation, and screening the fraction the inhibition rate of which is higher than 80%; and 4) reacting the fraction screened in the step 3) and the His-DXR obtained in the step 2), calculating the inhibition rate, and screening the fraction the inhibition rate of which is not less than 50%. The method is simple and accurate, and lays a foundation for development of novel antibacterial medicines adopting Dxr as a therapeutic target or development of leading compounds.