Food-borne lactobacillus for improving tobacco leaf quality, and preparation method and application thereof
A food-derived, tobacco leaf technology, applied in the field of applied microorganisms, can solve problems such as potential safety hazards, and achieve the effects of soft smoke, coordinated chemical components, and weakened bitter and spicy taste
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Embodiment 1
[0020] 1. Isolation and identification of Lactobacillus plantarum L15
[0021] Several tempeh samples were collected from various prefectures and counties in Yunnan, and the modified MRS solid medium was used to screen lactic acid bacteria. After a single colony was allowed to stand overnight in the MRS liquid medium, it was stored in a refrigerator at -80℃. Six strains of lactic acid bacteria were randomly selected, streaked and separated in tobacco leaf medium, and cultured at 37°C for 48 hours. One strain with the best vigor was selected, named L15, inoculated on MRS solid medium, cultured at 37°C for 48 hours, the colony morphology was observed, and physiological and biochemical characteristics were analyzed according to conventional bacterial identification methods. Use bacterial genome extraction kit to extract L15 genomic DNA, use 16S rDNA universal primer 27F
[0022] (5’-AGAGTTTGATCCTGGCTCAG-3’) and 1492R
[0023] (5’-GGTTACCTTGTTACGACTT-3’) amplified L15 16S rDNA gene and...
Embodiment 2
[0055] Application of Lactobacillus plantarum L15
[0056] 1) Preparation of fermentation inoculum: Inoculate Lactobacillus plantarum L15 activated in MRS medium for 16 hours in 100 mL MRS liquid medium at 8‰ (v / v) inoculation amount, and cultivate at 37°C for 24 hours at 8000 rpm , 25min centrifugation to pellet the bacteria, wash the bacteria with sterile deionized water for 2-3 times, add sterile deionized water to make the concentration 10 10 cfu / mL bacterial suspension, this is Lactobacillus plantarum L15 fermentation inoculum.
[0057] 2) Take out tobacco shreds from the commercial Yunyan Ziyun, spray 6mL of Lactobacillus plantarum L15 fermentation inoculum into 20g shredded tobacco with a moisture content of 11%, and ferment in a constant temperature and humidity incubator at 28°C, 60% humidity, and 14 Today, cigarettes are made according to conventional methods for sensory evaluation. The control sample uses the same amount of sterile water instead of the fermentation inoc...
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