Modified beer waste yeast adsorbent, preparation method and application thereof
A technology for beer waste yeast and adsorbent, which is applied in biochemical equipment and methods, chemical instruments and methods, enzymes, etc., can solve problems such as high cost and complexity, and achieve low cost, good vitality yield, and biological activity maintenance. good effect
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Embodiment 1
[0039] A modified beer waste yeast adsorbent, the steps of its preparation method are as follows:
[0040] 1. Pretreatment of waste beer yeast: Weigh 15.0g waste beer yeast into a 250mL Erlenmeyer flask, add 150mL distilled water, shake in a water bath at 25°C for 1 hour, take it out and let it stand for 30 minutes, discard the upper layer of turbid liquid, and continue to wash the waste beer yeast with water Until the supernatant is colorless, wash with absolute ethanol for 3 times, let stand, and then filter with suction to separate the solid from the liquid, and place the obtained solid in an oven at 60°C to dry and set aside;
[0041] 2. Preparation of cross-linked waste beer yeast: add 1.5 g of waste beer yeast pretreated in step 1 into 50 mL of 2% wt glutaraldehyde solution, shake the reaction at 35°C for 12 hours and collect the product. Washing with water three times, and then washing with absolute ethanol three times to remove unreacted glutaraldehyde on the surface o...
Embodiment 2
[0044] Embodiment 2: apply the adsorbent prepared by the method of the present invention to adsorb lysozyme
[0045] Determination of adsorption conditions:
[0046] 1. Determination of adsorption acidity: Weigh 0.0200g of cross-linked beer waste yeast and modified beer waste yeast adsorbent (weigh multiple parts in parallel), place them in 50mL Erlenmeyer flasks respectively, add 10.00mL of 0.2mg·mL -1 Lysozyme standard solutions with different acidities (pH=3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.4, 8.0), adsorbed on a shaker at 120 rpm at 25°C for 12 hours, took out the supernatant, measured, and calculated The adsorption rate of adsorbent to lysozyme, and its adsorption acidity curve, see Figure 4 ;
[0047] For the modified beer waste yeast, with the increase of pH, the adsorption rate of lysozyme increases gradually. For higher activity, pH 7.4 was selected as the experimental pH of the following examples. For cross-linked beer waste yeast, when pH<4, lysozyme doe...
Embodiment 3
[0053] Example 3: Regeneration Performance Test of Modified Beer Spent Yeast Adsorbent
[0054] 1. Take 0.0200g of modified yeast into a 50.0mL Erlenmeyer flask, add 10.00mL of 0.2mg·mL prepared with pH7.4 buffer solution (0.05M Tris-hydrochloric acid system) -1 Put the lysozyme standard solution into a water-bath shaker at 25°C, and adsorb on the shaker at 120rpm for 40min. After standing still, take out the supernatant and measure the adsorption rate of lysozyme (at this time, it is the adsorption rate for the first use). After the remaining adsorbent is naturally air-dried, add 10.00mL of 1.0M NaCl solution, put it in a water-bath shaker at 25°C, and elute on the shaker at 120rpm for 40min. After standing still, take out the supernatant and wait for the remaining adsorbent Repeat the above adsorption-elution after natural air-drying, the adsorbent was used four times in total, and the adsorption capacity of the fourth use still maintained 94.6% of the first use (see Table 4...
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