Preparation method and application of Bacillus methylotrophicus and tobacco spine date seed extract thereof
A technology of methanol-eating spores and sour jujube kernels, which is applied in the field of preparation of Bacillus methanol-eating and sour jujube seed extracts for cigarettes, can solve the problems of environmental pollution, single aroma and taste of tobacco extracts, insufficient aroma intensity, etc. The effect of wide availability, simple process and wide source of raw materials
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Embodiment 1
[0035]The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of Zianzaorex extractum for tobacco as follows:
[0036] 1) The jujube seed and water were refluxed and extracted at a mass ratio of 1:20 for 3 hours, then filtered, and the filtrate was sterilized at 115°C for 25 minutes;
[0037] 2) Slant culture: inoculate Bacillus methanolivorus 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;
[0038] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0039] 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;
[0040] The composition of the seed liquid medium is: maltose 50g / L, peptone 10g / L, sodiu...
Embodiment 2
[0044] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of Zianzaorex extractum for tobacco as follows:
[0045] 1) The jujube seed and water were refluxed and extracted at a mass ratio of 1:30 for 2 hours, then filtered, and the filtrate was sterilized at 115°C for 25 minutes;
[0046] 2) Slant culture: inoculate Bacillus methanolivorus on the slant medium of potato dextrose agar, at 30°C and pH 7, let it stand for 36 hours and place it in the refrigerator;
[0047] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0048] 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;
[0049] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, sodium chloride 5g / L;
[0...
Embodiment 3
[0053] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of Zianzaorex extractum for tobacco as follows:
[0054] 1) The jujube seed and water were refluxed and extracted at a mass ratio of 1:40 for 1 hour, then filtered, and the filtrate was sterilized at 115°C for 25 minutes;
[0055] 2) Slant culture: inoculate Bacillus methanolivorus on the slant medium of potato dextrose agar, at 35°C and pH 5, culture it statically for 72 hours and then place it in the refrigerator;
[0056] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0057] 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;
[0058] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, sodium chl...
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