A kind of bio-targeted antibacterial dspb immobilizing enzyme and its preparation method and application
A technology for immobilizing enzymes and organisms, applied to biochemical equipment and methods, and enzymes immobilized on or in inorganic carriers, can solve the problems of low stability and activity of glycoside hydrolases, and achieve easy enrichment and separation, The effect of good dispersion and high recycling efficiency
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Embodiment 1
[0037] A preparation method for biological targeting antibacterial DspB immobilizing enzyme, comprising the following steps:
[0038] 1) Preparation of magnetic nanomaterials: Weigh 2.33g FeCl 3 ·6H 2 O and 1.9g Fe(NH 4 ) 2 ·(SO 4 ) 2 ·6H 2 O, dissolved together in 50ml deoxygenated deionized water, and quickly added NH during stirring at 60°C 3 ·H 2 O to adjust the pH to 9-12, closed reaction for 30min, and then aging at 80°C for 0.5-2.0h to obtain superparamagnetic Fe 3 o 4 nanoparticles, and in order to improve the Fe 3 o 4 The purity of the nanoparticles, thereby improving the efficiency of the later reaction, will prepare the Fe 3 o 4 Nanoparticles are washed to neutrality after magnetic separation;
[0039] Take 200mg Fe 3 o 4 Suspend nanoparticles in 40mL deionized water and 160mL absolute ethanol, add 6mL concentrated ammonia water after ultrasonic dispersion, and slowly add 100-1000μL tetraethyl orthosilicate (TEOS) under mechanical stirring at 25°C, an...
Embodiment 2
[0044] The difference between this example and Example 1 is that in this example, the modification of the magnetic nanomaterials uses dopamine for polydopamine modification.
[0045] The modification of magnetic nanomaterials in this embodiment specifically includes:
[0046] Dissolve 0.121g Tris-base in 100mL deionized water, adjust the pH to 8.5 with hydrochloric acid to obtain Tris-HCl buffer;
[0047] Add 0.1-1.0g of dopamine to the Tris-HCl buffer, take 200mg of magnetic nanomaterial A to absorb and remove the supernatant, add it to the Tris-HCl buffer, ultrasonically disperse for 10-30min, and stir the reaction in a water bath at 25°C for 10- 18h, carry out polydopamine modification reaction, after polydopamine modification reaction finishes, magnetic adsorption removes water, obtains the magnetic nano material B of immobilizing glycoside hydrolase, the end of this magnetic nano material B is modified with polydopamine (Fe 3 o 4 -SiO 2 -PDA), and in order to improve t...
Embodiment 3
[0050] The difference between this example and Example 1 is that the modification of the magnetic nanomaterial in this example uses succinic anhydride for carboxyl modification.
[0051] The modification of magnetic nanomaterials in this embodiment specifically includes:
[0052] Dissolve 100mg of magnetic nanomaterial A in absolute ethanol and disperse it ultrasonically. Slowly add 0.1-10mL of γ-aminopropyltriethoxysilane dropwise under mechanical stirring, and then reflux and condense at 80°C for 4-8h. Amination modification reaction was carried out to obtain amino-modified nanomaterials (Fe 3 o 4 -SiO 2 -APTES);
[0053] Take 100mg of amino-modified nanomaterials, wash them with absolute ethanol for several times, ultrasonically oscillate for 10-30min each time, and suspend in an appropriate amount of absolute ethanol;
[0054] Weigh 2.0-5.0 g of succinic anhydride, dissolve it in an appropriate amount of dimethyl sulfoxide (DMSO), and then, under vigorous stirring, mix...
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