Embedding-crosslinking method for preparing nicotinic acid by Gibberella immobilization bioconversion
A technology for biotransformation, Gibberella, applied in biochemical equipment and methods, microorganism-based methods, immobilized on/in organic carriers, etc., can solve the problems of active damage, high glutaraldehyde toxicity, etc.
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example 1
[0013] Glutaraldehyde cross-linked immobilized cells
[0014] The method of immobilizing cells with polyvinyl alcohol-chitosan composite materials: the prepared bacterial liquid is centrifuged to obtain wet bacterial cells, after centrifugal washing with normal saline, 0.05 g of bacterial cells are suspended in normal saline to prepare 10 g / L bacterial suspension, add polyvinyl alcohol solution with a mass fraction of 3% and chitosan solution with a mass fraction of 6%, mix thoroughly and uniformly according to a certain proportion, and then inject it with a syringe steadily into a solution containing 8% (w / v) polysaccharide Sodium phosphate saturated boric acid solution was solidified at 0°C for 6 hours to obtain immobilized pellets, which were filtered through gauze and then washed with PBS buffer to remove unfixed cells and mixed with the solidification solution. After curing, the pellets were stored in normal saline. The immobilized pellets are milky white and about 2mm i...
example 2
[0024] Genipin cross-linked immobilized cells
[0025] According to the method in Example 1, an equal amount of 7 parts of two kinds of immobilized cells were respectively prepared, and each part of the immobilized cells was treated with different concentrations of genipin for 12 hours, and the treated immobilized cells were washed with physiological saline after cross-linking, and then The immobilized cells were placed in saline and stored at 4°C.
[0026] The conditions for determining the viability were as follows: the immobilized cells cross-linked by genipin were placed in 7 mL of PBS buffer solution with a pH of 7.4, and 7 mL of 100 mM 3-cyanopyridine was added thereto to form a 14 mL reaction system for 30 min. After the reaction was completed, the liquid phase was used to detect the conversion result.
[0027] Table 3 Different concentrations of genipin treated polyvinyl alcohol-chitosan immobilized cells
[0028]
[0029] Table 4 Sodium alginate-gelatin immobiliz...
example 3
[0033] Batch conversion of niacin using two immobilized cells treated with two crosslinkers
[0034] Batch experiments were carried out with 6 mL of two kinds of immobilized cells treated with a certain amount of genipin and glutaraldehyde, and then 7 mL of PBS buffer was added to it, and 7 mL of substrate 3-cyanopyridine solution with a concentration of 100 mM was added, and the temperature was 40 °C. , react for 40min. At the end of each batch of transformation, the immobilized cells were washed with PBS buffer three times and then put into a new transformation system for the next batch of reactions.
[0035] Table 5 Batch transformation comparison of glutaraldehyde crosslinked immobilized cells and genipin immobilized cells
[0036]
[0037] It can be seen from Table 5 that the sodium alginate-gelatin immobilized cells without cross-linking treatment can only carry out 17 batches of reactions, and the cells immobilized by glutaraldehyde cross-linking can be reused for 2...
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