Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof
A technology of methanol-eating spores and bacilli, which is applied in the field of preparation of methanol-eating bacillus and its chaff extract for tobacco, can solve the problems of single aroma and taste of tobacco extract, insufficient aroma intensity, and complicated extraction steps, and achieve increased Characteristic aroma, rich smoke aroma, low cost of raw materials
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Embodiment 1
[0032]The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of chaff extract for tobacco as follows:
[0033] 1) The chaff 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;
[0034] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1 CCTCCNO: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;
[0035] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0036] 3) Preparation of seed culture medium: use the slant culture obtained in the above step 2) to inoculate the seed culture medium, vibrate at 25°C and rotate at 150rpm for 12 hours, and transfer 4 times in succession to prepare the seed culture medium;
[0037] The composition of the seed liquid medium is: maltose 50g / L, peptone ...
Embodiment 2
[0041] The present invention utilizes microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of chaff extract for tobacco as follows:
[0042] 1) The husk 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;
[0043] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCCNO:M 2012004 on the potato dextrose agar slant medium, and culture it statically for 36 hours at 30°C and pH 7, then place it in the refrigerator;
[0044] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0045] 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;
[0046] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, sodium ch...
Embodiment 3
[0050] The present invention utilizes the microorganism Bacillus methylotrophicus VJ4-1CCTCC NO: M 2012004 to biotransform the preparation method of chaff extract for tobacco as follows:
[0051] 1) The husk 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;
[0052] 2) Slant culture: inoculate Bacillus methylotrophicus VJ4-1CCTCCNO: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;
[0053] Slant medium: potato infusion 200mL / L, glucose 20g / L, agar 13g / L;
[0054] 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;
[0055] The seed liquid medium consists of: maltose 50g / L, peptone 10g / L, so...
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