Immobilization method of free enzyme
A lipase and metal salt technology, applied in the field of immobilization of free enzymes, can solve problems such as poor structural stability, difficult separation of free enzymes, narrow particle size distribution, etc., achieve low mechanical loss rate, overcome crystal transformation, and maintain enzyme activity high rate effect
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Embodiment 1
[0030] (1) Two parts of identical microemulsions were prepared at 25°C, each part of microemulsion consisted of: 100g n-hexane / ethyl acetate (n-hexane 33.3g, ethyl acetate 66.7g), 100g dodecylaminopropionic acid and 100g deionized water.
[0031] (2) Weigh 10 g of CALB lipase dry powder, add 50 mL, 0.2 mol / L sodium hydrogen phosphate buffer solution therein, then add 10 g of barium chloride powder therein, and stir to obtain a white emulsion. The obtained white emulsion was added to one part of the microemulsion prepared in step (1), and it was used under stirring state, which was recorded as the microemulsion mixed liquid one.
[0032] (3) Prepare 100g mass fraction of 10wt% sodium sulfate aqueous solution, add it to the above-mentioned microemulsion two, then add 10g triethylamine therein, stir and mix it for later use, and record it as microemulsion mixed solution two.
[0033] (4) Add two drops of the microemulsion mixed solution in step (3) to the first microemulsion mix...
Embodiment 2
[0036] (1) Prepare two identical microemulsions at 25°C, each microemulsion consists of: 100g n-hexane / ethyl acetate (n-hexane 50.0g, ethyl acetate 50.0g), 50g dodecylaminopropionic acid and 100g deionized water.
[0037] (2) Weigh 10g of CALB lipase dry powder, add 60mL, 0.4mol / L sodium hydrogen phosphate buffer solution to it, then add 20g of barium chloride powder to it, and stir to obtain a white emulsion. The obtained white emulsion was added to one part of the microemulsion prepared in step (1), and it was used under stirring state, which was recorded as the microemulsion mixed liquid one.
[0038] (3) Prepare 100g mass fraction of 12wt% sodium sulfate aqueous solution, add it to the above-mentioned microemulsion two, then add 20g triethylamine therein, stir and mix it for later use, and record it as microemulsion mixed liquid two.
[0039] (4) Add two drops of the microemulsion mixed solution in step (3) to the first microemulsion mixed solution in step (2) at 15°C, an...
Embodiment 3
[0042] (1) Two identical microemulsions were prepared at 25° C., each microemulsion consisted of: 100 g of n-hexane, 100 g of dodecylalanine and 100 g of deionized water.
[0043] (2) Weigh 10 g of CALB lipase dry powder, add 50 mL, 0.2 mol / L sodium hydrogen phosphate buffer solution therein, then add 10 g of barium chloride powder therein, and stir to obtain a white emulsion. The obtained white emulsion was added to one part of the microemulsion prepared in step (1), and it was used under stirring state, which was recorded as the microemulsion mixed liquid one.
[0044] (3) Prepare 100 g of sodium sulfate aqueous solution with a mass fraction of 10 wt %, add it to the above-mentioned microemulsion two, then add 10 ethylenediamine therein, stir and mix it for later use, and record it as microemulsion mixed liquid two.
[0045] (4) Add two drops of the microemulsion mixed solution in step (3) to the first microemulsion mixed solution in step (2) at 10°C, control the dropping ti...
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