Organic-inorganic composite immobilized cell and its preparation method and application
A technology of immobilized cells and inorganic composites, applied in biochemical equipment and methods, immobilized enzymes, immobilized on or in inorganic carriers, etc., to achieve the effects of low cost, enhanced tolerance, and excellent stability
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Embodiment 1
[0036] The preparation of embodiment 1 bacterial suspension
[0037] The nitrilase-producing mutant mut-D3 was inoculated in 50 mL of liquid medium, and cultured with shaking at 30° C. for 20-36 h, preferably 24 h in this embodiment, as the seed liquid. Transfer to fresh fermentation medium with 1-5% inoculum size and shake culture at 30°C for 30-45h. In this embodiment, the preferred inoculum size is 1%, and the culture time is 40 h. Centrifuge at 8000 rpm for 10 min at 4°C to remove the supernatant and collect the cells. Use 100mM, pH7.2 sodium phosphate buffer to wash 2-3 times, preferably 2 times in this embodiment, and resuspend the bacterium in the buffer to make a 20mg / mL bacterial suspension, place in 4 ℃ refrigerator for later use.
Embodiment 2
[0038] Example 2 Preparation of organic-inorganic composite immobilized cells
[0039] This implementation provides a method for preparing organic-inorganic composite immobilized cells, adding inorganic materials SiO2 to sodium alginate-PVA immobilized cells 2 , CaCO 3 , bentonite, to investigate the influence of a single inorganic material on the performance of immobilized cells, the specific method is as follows:
[0040] The sodium alginate aqueous solution with a mass concentration of 2.5% and the PVA aqueous solution with a mass concentration of 8% were mixed according to a ratio (volume ratio) of 9:1 as an organic fixing material. The above-mentioned organic fixing material was mixed with the bacterial suspension prepared in Example 1 (volume ratio) at 1:1 to obtain a composite organic material solution. Then add different inorganic materials to the composite organic material solution: SiO 2 (1.0%, 1.5%, 2.0%, 2.5%, 3.0%), CaCO 3 (0.2%, 0.4%, 0.6%, 0.8%, 1.0%), bento...
Embodiment 3
[0043] Example 3 Preparation of organic-inorganic composite immobilized cells
[0044] This implementation provides a preparation method of organic-inorganic composite immobilized cells, adding different concentrations of inorganic material SiO to sodium alginate-PVA immobilized cells at the same time 2 , CaCO 3 , to investigate the effect of composite inorganic materials on immobilized cells, the specific method is as follows:
[0045] The sodium alginate aqueous solution with a mass concentration of 2.5% and the PVA aqueous solution with a mass concentration of 8% were mixed according to a ratio (volume ratio) of 9:1 as an organic fixing material. The above-mentioned organic fixing material was mixed with the bacterial suspension prepared in Example 1 (volume ratio) at 1:1 to obtain a composite organic material solution. Add 1.5% SiO to the composite organic material solution 2 +0.6%CaCO 3 , 1.5% SiO 2 +0.8%CaCO 3 , 1.5% SiO 2 +1.0%CaCO 3 , 2.0% SiO 2 +0.6%CaCO 3 ,...
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