Method for extracting laccase from pleurotus eryngii fungi residues
A technology of Pleurotus eryngii bacteria residue and laccase, which is applied in the biological field, can solve the problems of limited production, extraction of laccase, and high production cost, and achieve the effects of improving utilization rate, increasing specific activity, and easy operation
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Embodiment 1
[0024] A kind of method of present embodiment extracts laccase from Pleurotus eryngii fungus slag, concrete process is as follows:
[0025] Select fresh fungus residues that have just harvested fruit bodies, crush these fungus residues, and pass through a 4-mesh sieve; then add an extract to the crushed fungal residues, the extract used is HAc-NaAc with a pH of 4.5 to 5.5 Buffer solution, add according to the ratio of material to liquid 1g: 5ml, after soaking for 24 hours, filter to remove solids, centrifuge at 4800r / min for 10min, collect the supernatant as the crude enzyme solution, this method is simple, the extraction rate is high, and it is convenient for subsequent collection of crude enzyme Enzyme solution. Then use the tangential flow ultrafiltration system to concentrate the crude enzyme solution. Since the molecular weight of laccase is about 120KD, the crude enzyme solution is first filtered with a 50KD tangential flow ultrafiltration membrane to intercept some macr...
Embodiment 2
[0031] A kind of method of present embodiment extracts laccase from Pleurotus eryngii fungus slag, concrete process is as follows:
[0032] Select the fungal residues that have been placed in a cool place within 3 days, crush these fungal residues, and pass through a sieve of about 10 meshes; then add an extract to the crushed fungal residues, the extract used is HAc- Add NaAc buffer solution according to the ratio of material to liquid: 1g: 10ml. After soaking for 12 hours, filter to remove solids, centrifuge at 5000r / min for 5 minutes, and collect the supernatant as the crude enzyme solution. Then use the tangential flow ultrafiltration system to concentrate the crude enzyme solution. The crude enzyme solution is first filtered with a 50KD tangential flow ultrafiltration membrane to intercept some macromolecular impurities, collect the permeate, and then use a 30KD tangential flow ultrafiltration membrane to filter the crude enzyme solution. The ultrafiltration membrane conc...
Embodiment 3
[0035] This example is a method for extracting laccase from Pleurotus eryngii fungus dregs. The specific process is the same as in the example, except that the solid-liquid ratio of fungus dregs to extract is 1g:8.2ml, soaked for 36 hours.
[0036] The characterization results of the laccase enzyme powder obtained by the above method are as follows: the specific activity is 9400U / g, and the enzyme activity is stable at a temperature of 20-60°C; at room temperature, the change of the enzyme activity within the pH range of 4.0-6.6 is less than 11.7%.
[0037] It should be noted that when a numerical range is involved in the claims of the present invention, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. 3 Similarly, in order to avoid repetition, the present invention describes preferred embodiments, but those skilled in the art can make additional changes and modifications to these embodiments once th...
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Abstract
Description
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