Method for degrading sterol compounds in tobacco through microorganism combination method
A technology of microbial degradation and sterols, which is applied in the fields of tobacco, tobacco processing, food science, etc., can solve the problems of slow growth rate and low efficiency of sterol degradation, and achieve the effect of improving tolerance
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Embodiment 1
[0024] Example 1. Cultivation of Arthrobacter and Mycobacterium
[0025] 1.1 Arthrobacter
[0026] (1) Arthrobacter growth medium: beef extract 5g, egg white 10g, NaCl 5g, nicotine 2g, agar 15g, add water to make the volume to 1L, adjust the pH to 7.0, and sterilize at 121°C for 20min.
[0027] (2) Arthrobacter strain: Inoculate the seed solution of Arthrobacter into the growth medium with 10% inoculum, and cultivate at 30°C and 200rpm for 24 hours to obtain the Arthrobacter strain. The concentration of Arthrobacter in the strain is 10 9 Pieces / mL.
[0028] 1.2 Mycobacterium
[0029] (1) Mycobacterium liquid growth medium: glucose 10.0g, K 2 HPO 4 0.5g, MgSO 4 ·7H 2 O 0.5g, NH 4 NO 3 2g, 0.05g ferric amine citrate, 2.0g citric acid, 5.0g glycerol, add water to make the volume to 1L, adjust the pH to 7.0-7.2, and sterilize at 121℃ for 20min (prepare solid plate medium and add liquid growth medium mass fraction 2% agar).
[0030] (2) Mycobacterium resting cell liquid: inoculate cryoprese...
Embodiment 2
[0032] The method for degrading sterols in tobacco by the microbial combination method of the present invention includes the following steps:
[0033] (1) Take 30g each of the tobacco stems and dust, extract 20 times the weight of water at 55°C for 1 hour, and then pass through a 200-mesh screen for solid-liquid separation, and the resulting liquid is the tobacco extract;
[0034] (2) According to 10% of the inoculation amount, add Arthrobacter bacterium liquid to the tobacco extract and ferment at 30°C and 200 rpm to reduce the nicotine content in the tobacco extract so that the nicotine content is less than 0.5g / L;
[0035] (3) Add the resting cell liquid of mycobacteria to the tobacco extract of step (2) according to the 10% inoculum, and ferment at 37°C and 200 rpm for 48 hours to reduce the sterols in the tobacco extract content.
Embodiment 3
[0037] The method for degrading sterols in tobacco by the microbial combination method of the present invention includes the following steps:
[0038] (1) Take 30 g of tobacco leaves, use 30 times the weight of water, extract for 70 minutes at 50°C, and then pass through a 200 mesh screen for solid-liquid separation, and the obtained liquid is the tobacco extract;
[0039] (2) According to the inoculation amount of 12%, add Arthrobacter bacteria liquid to the tobacco extract, and ferment at 25°C and 250rpm to reduce the nicotine content in the tobacco extract so that the nicotine content is less than 0.5g / L;
[0040] (3) According to the inoculation amount of 12%, add the resting cell liquid of mycobacteria to the tobacco extract of step (2), and ferment at 35°C and 250 rpm for 50 hours to reduce the sterols in the tobacco extract content.
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