Method for preparing endogenous total enzyme by using fresh tobacco and application of endogenous total enzyme
An endogenous, tobacco technology, applied in the application, tobacco, treatment of tobacco and other directions, can solve problems such as unreported, achieve the effects of reasonable process, promotion of tobacco leaf alcoholization, good economic and social benefits
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Embodiment 1
[0025] The method for preparing endogenous total enzymes from fresh tobacco comprises the following steps:
[0026] (1) Raw material treatment: Wash the discarded fresh tobacco raw materials, squeeze out the juice with a press, quickly filter with gauze to remove suspended plant particles, and then use a centrifuge at 3000r / min for 60min to remove impurities, and collect the supernatant for later use ;
[0027] (2) Concentration by ultrafiltration: Concentrate the supernatant in ultrafiltration equipment, the molecular weight cut-off (MWCO) of the ultrafiltration membrane is 10kD, the temperature is 4°C, and the pressure is 0.10MPa, so that the liquid volume is concentrated to about 20% of the original volume;
[0028] (3) Precipitation and refinement: cool the concentrated solution to 0-2°C, add edible ethanol with a mass concentration of 95% while stirring until the mass concentration of ethanol in the mixed solution is 10%, leave it to stand to precipitate the enzyme, remov...
Embodiment 2
[0033] The method of using fresh tobacco to prepare endogenous total enzymes is to wash the discarded fresh tobacco raw materials, squeeze out the juice with a press, quickly filter out suspended plant particles with gauze, and then use a centrifuge at 15000r / min for 10min to remove impurities and collect Serum for later use. Concentrate the supernatant in an ultrafiltration device with an ultrafiltration membrane molecular weight cut-off (MWCO) of 50kD, a temperature of 35°C, and a pressure of 0.40MPa to concentrate the liquid volume to 19% of the original volume. Cool the concentrated solution to 2-4°C, add edible ethanol while stirring until the mass concentration of ethanol in the mixed solution is 90%, let it stand to allow the enzyme to precipitate, remove the supernatant, collect the precipitate to obtain the wet enzyme. The wet enzyme is frozen at a low temperature below 0°C for 24 hours to be dried, and then pulverized to obtain the tobacco endogenous total enzyme pro...
Embodiment 3
[0037] The method of using fresh tobacco to prepare endogenous total enzymes is to wash the discarded fresh tobacco raw materials, squeeze out the juice with a press, quickly filter out suspended plant particles with gauze, and then use a centrifuge at 4000r / min for 30min to remove impurities and collect them. Serum for later use. Concentrate the supernatant in an ultrafiltration device with an ultrafiltration membrane molecular weight cut-off (MWCO) of 20kD, a temperature of 25°C, and a pressure of 0.20MPa to concentrate the liquid volume to 21% of the original volume. Cool the concentrated solution to 3°C, add ethanol while stirring until the mass concentration of ethanol in the mixed solution is 20%, let it stand still to allow the enzyme to precipitate, remove the supernatant, collect the precipitate to obtain the wet enzyme. The wet enzyme is frozen at a low temperature below 0°C for 4 hours, dried, and then pulverized to obtain the tobacco endogenous total enzyme product...
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