A kind of preparation method and application of the silk nano ball of encapsulation enzyme
A nano ball and silk technology, applied in the field of enzyme encapsulation, can solve problems such as poor biocompatibility and unfavorable use in animals, and achieve the effect of protecting enzyme activity, protecting storage stability and ensuring stability.
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Embodiment 1
[0037] Example 1 Preparation of Enzyme-Coated Silk Nanospheres
[0038] A method for preparing enzyme-encapsulated silk nanospheres, comprising the following steps:
[0039] S1, prepare silk solution, specifically:
[0040] S11, shred silkworm cocoons and place them in 5% NaHCO 3 Boil in the solution, and stir with a glass rod for 30min, repeat this step twice;
[0041] S12. Wash the silk obtained in step S11 with 60°C deionized water for 20 minutes, and repeat this step three times;
[0042] S13, drying the washed silk in an oven at 60°C, then dissolving it in a 9.3M LiBr solution, and dissolving at 60°C for 4 hours;
[0043] S14, dialyze the silk solution obtained in step S13 with deionized water for two days, and change the water every 2 hours;
[0044] S15. Dilute the silk solution obtained in step S14 to 6% w / w;
[0045] S2, encapsulating the enzyme into silk to form silk nanospheres encapsulating the enzyme, specifically:
[0046] S21. Take 1ml of the silk solution...
Embodiment 2
[0053] Example 2 Determination of the activity of enzymes wrapped in silk nanospheres
[0054] (1) Thermostability and storage stability of the enzyme
[0055] By measuring the relative activity of the enzyme without silk wrapping and the enzyme wrapped with silk at different time points, the heat stability and storage stability of the enzyme before and after wrapping were compared. The result is as figure 2 shown.
[0056] figure 2a represents the relative enzymatic activity of glucose oxidase without silk wrapping and silk wrapping glucose oxidase at 60°C. It can be seen from the figure that at 60°C, glucose oxidase without silk wrapping loses half of its activity in 1 hour; while glucose oxidase wrapped in silk loses half of its activity after 30 hours; The thermal stability of the enzyme encapsulated by nanospheres is effectively protected.
[0057] figure 2 b represents the storage stability of horseradish peroxidase without silk wrapping and silk wrapping horser...
Embodiment 3
[0067] Embodiment 3 in the application of antialcoholic medicine
[0068] Alcohol oxidase (AOx) and catalase (Cat) were wrapped in silk nanospheres to prepare a protease-resistant anti-alcoholic drug, and its effect was as follows: Figure 6 shown.
[0069] In the simulated intestinal fluid, add the alcohol enzyme without silk nanospheres or the alcoholase with silk nanospheres, and compare the relative enzyme activities of the two, as shown in Figure 6 As shown in a, the alcohol enzyme wrapped with silk nanospheres still retains more than 90% of its activity after 2 hours, while the relative activity of the alcoholase without silk nanospheres wrapped rapidly drops to 5% after 20 minutes. This is because the drug containing protease reaches the intestinal juice after oral administration. The protease in the intestinal juice will promote the hydrolysis of protein, and the essence of most enzymes is protein, which leads to the rapid decomposition and inactivation of the enzyme...
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