Bacillus methylotrophicus and preparation method and application of tobacco coffee extract thereof
A technology of methanol-eating bacillus and bacillus, which is applied in the field of preparation of methanol-eating bacillus and its coffee extract for tobacco, can solve the problems of various types of dominant bacteria, difficult control of production conditions, unsuitable for large-scale industrial production, etc., and achieve industrialization Conditions are easy to control, reduce miscellaneous air, increase the effect of delicate and soft feeling
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Embodiment 1
[0033] The present invention utilizes the microorganism Bacillus methylotrophicus (Bacillus methylotrophicus) VJ4-1 CCTCC NO: M 2012004 to bioconvert the coffee extract for tobacco and prepare the method as follows:
[0034]1) Crush the coffee beans to 10 meshes, reflux extraction with water at a mass ratio of 1:20 for 3 hours, then filter, and sterilize the filtrate at 115°C for 25 minutes;
[0035] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1 CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 24 hours at 25°C and pH 10, then place it in the refrigerator;
[0036] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0037] 3) Preparation of seed culture medium: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate for 12 hours at 25°C and rotate at 150 rpm, and transfer 4 times in succession to prepare the seed culture medium;
[0038] The composition of the see...
Embodiment 2
[0042] The present invention utilizes the microorganism Bacillus methylotrophicus (Bacillus methylotrophicus) VJ4-1 CCTCC NO: M 2012004 to bioconvert the coffee extract for tobacco and prepare the method as follows:
[0043] 1) Crush coffee beans to 100 mesh, reflux extraction with water at a mass ratio of 1:30 for 2 hours, filter, and sterilize the filtrate at 115°C for 25 minutes;
[0044] 2) Slant culture: inoculate Bacillus methanolivorus on the potato dextrose agar slant medium, and culture it for 36 hours at 30°C and pH 7, then place it in the refrigerator;
[0045] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0046] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed medium, vibrate and culture at 30°C and 120 rpm for 24 hours, and make the seed culture solution after continuous transfer for 3 times;
[0047] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, sodium chl...
Embodiment 3
[0051] The present invention utilizes the microorganism Bacillus methylotrophicus (Bacillus methylotrophicus) VJ4-1 CCTCC NO: M 2012004 to bioconvert the coffee extract for tobacco and prepare the method as follows:
[0052] 1) Crush coffee beans to 200 mesh, reflux extraction with water at a mass ratio of 1:40 for 1 hour, then filter, and sterilize the filtrate at 115°C for 25 minutes;
[0053] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1 CCTCC NO: M 2012004 on the potato dextrose agar slant medium, and culture it statically for 72 hours at 35°C and pH 5, then place it in the refrigerator ;
[0054] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0055] 3) Preparation of seed culture solution: use the slant culture obtained in the above step 2) to inoculate the seed medium, vibrate and culture at 35°C and rotate at 100 rpm for 36 hours, and make the seed culture solution after two consecutive transfers;
[0056] The seed liquid medium cons...
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