A kind of enzyme preparation using nicotine dehydrogenase as auxiliary tobacco processing and its preparation and use method
A technology of nicotine dehydrogenase and alkali dehydrogenase, which is applied in the field of enzyme preparations to improve tobacco grades, can solve the problems of affecting tobacco leaf smoking evaluation results and low nicotine content, and achieve the effect of easy use and reduced nicotine content
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[0041] (1) Preparation and determination of the preparation of the present invention
[0042] Nicotine dehydrogenase can be purchased directly from microbial fermentation products. Or use the corresponding nicotine dehydrogenase-producing strains to ferment and produce crude products through cell wall breaking, release and preliminary extraction. Then, refine, remove metal ions, dry and concentrate with the method of the present invention to obtain the nicotine dehydrogenation preparation.
[0043] Dissolve and dilute the solid nicotine dehydrogenase crude product with W / V=1g:20mL or liquid W / V=1mL:10mL buffer, the pH value of Britton-Robinsion buffer is 7.0; centrifuge at 3500r / min for 20min, take the Clear solution, add (NH 4 ) 2 SO 4 , so that the final concentration of ammonium sulfate is 62%, let it stand for 24h, then centrifuge at 15000r / min at 4°C for 30min, take the supernatant, and add (NH 4 ) 2 SO 4 Overnight, centrifuge again at 15000r / min ...
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