Process for preparing membrane used to immobilize microbes to biosense biochemical oxygen demand
A technology of microbial immobilization and biochemical oxygen demand, applied in biochemical equipment and methods, microbial determination/inspection, biological material pretreatment, etc., can solve problems such as poor stability of BOD microbial membranes, and maintain activity and stability. , good adhesion, small diffusion resistance effect
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Embodiment 1
[0009] Example 1: Add tetramethoxysilane (TMOS) and dimethyldimethoxysilane (DiMe-DMOS) into a vial at a volume ratio of 1:1.2, shake fully and evenly, then stir and add the same solution as TMOS dropwise. The volume of 0.01mol / L HCl aqueous solution, the pH is 3, the mixed solution was stirred and hydrolyzed in a 60°C water bath for 2 hours, the mixed solution was milky white, and it was allowed to stand for 30 minutes, leaving a layer of gel solution for later use; prepare 8% (W / V) PVA-124 aqueous solution, after cooling, mix the bacterial suspension preserved with 0.067mol / L phosphate buffer (pH is 7.2) by 1:1 volume ratio, then add the above-mentioned gel solution of equal volume, Mix well and evenly, and spread microbial films of different thicknesses on the surface of glass substrates, oxidation sensing probes, etc.
[0010] The above-mentioned preparation process mainly has the following characteristics: 1) No alcohol is added to the initial reaction solution, and the ...
Embodiment 2
[0011] Example 2: Using Hansennula anomala as the working strain, the above-mentioned preparation method was used to prepare a microbial immobilized membrane. TMOS:DiMe-DMOS was mixed with HCl aqueous solution (pH 6) at a volume ratio of 1:0.5. The mixed solution was stirred and hydrolyzed in a water bath at 50°C for 2.5 hours. After the mixed solution was milky white, it was allowed to stand at room temperature. The mixed solution was divided into upper and lower layers, and the supernatant was removed. Warm water can be used to prepare the PVA-124 aqueous solution, and the prepared microbial film is directly cured on the surface of the oxygen sensing film, and the oxygen sensing film is used for secondary sensing to detect the change of the dissolved oxygen concentration, so as to investigate the preparation of the method. Response properties of microbial immobilized membranes. The oxygen sensing membrane used uses 4,7-diphenyl-1,10-phenanthroline ruthenium as a fluorescent ...
Embodiment 3
[0012] Example 3: The TMOS:DiMe-DMOS is configured at a volume ratio of 1:1.8, and the resulting microbial film can be placed in a refrigerator at 4° C. for about 2 days and then dried to shape. Glucose-glutamic acid (GGA) solution with a BOD value of 100 mg / L can be added to continue storage at 4°C in the refrigerator. It is found through experiments that the microbial immobilized membrane prepared by the above method has the characteristics of sensitive and rapid response to glucose-glutamic acid (GGA) standard solution, and usually the time to complete the measurement of a GGA standard solution is within 10 minutes. By changing the amount of bacteria embedded in the microbial film, the linear range of the concentration of different BOD values can be obtained. Under the same conditions, the reproducibility of the GGA solution with a BOD value of 100 mg / L was continuously measured 6 times, and the maximum change rate of the fluorescence intensity The relative deviation is 2...
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Abstract
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