The smxp-03 bacterial strain and application thereof for degrading protein in tobacco leaves
A SMXP-03, protein technology, applied in applications, microorganism-based methods, bacteria, etc., can solve the problem of not meeting the needs of high-quality tobacco raw materials, achieve good promotion and application value, reduce protein content, and short processing cycles. Effect
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Embodiment 1
[0025] This embodiment only briefly introduces the screening and identification process of the SMXP-03 strain as follows.
[0026] The SMXP-03 strain was originally isolated from red-cured tobacco leaves produced in Sanmenxia in 2013. The colony is round or irregular on the NA culture plate, milky white, dull, the surface of the colony is flat or irregular folds and raised, the surface is rough and not smooth, and the colony is opaque. The shape under the oil mirror is as figure 1 As shown, the bacteria can be observed to be evenly distributed, rod-shaped, and rarely chain-shaped, with a size of (0.7~0.9)×(1.6~2)μm.
[0027] Using the 16S rDNA gene fragment as a universal primer, the strain was subjected to PCR and cloning and sequencing, and a 1416bp sequence was obtained. Phylogenetic analysis showed that the strain SMXP-03 was closely related to Bacillus pumilus ( Bacillus pumilus ) has a homology of up to 99%, that is, the closest relative to Bacillus pumilus (such as ...
Embodiment 2
[0033] In order to use the SMXP-03 strain screened in Example 1 to degrade proteins in tobacco leaves, it is first necessary to prepare the strain into an easy-to-use bacterial agent. The specific preparation process is as follows.
[0034] (1) Activated strains: put the SMXP-03 strain on the NA plate, and cultivate it in a biochemical climate box at 37°C for 1-2 days;
[0035] (2) Preparation of seed solution: Scrape the bacteria cultured in step (1) in the ultra-clean workbench, inoculate in a 18×180 test tube containing 5mL liquid NA medium, and place on a shaker at 30°C and 150 rpm Overnight culture;
[0036] (3) Expansion culture: transfer the seed solution in step (2) to a sterile 250mL Erlenmeyer flask containing 100mL liquid NA medium according to 1 mL bacterial suspension / 100 ml NA medium (1% volume ratio) medium, 30°C, 150 rpm, cultured to OD 600 = about 2.0;
[0037] (4) Preparation of bacterial agent: Centrifuge the bacterial solution in step (3) at 3600 rpm for...
Embodiment 3
[0039] When utilizing bacterial agent prepared in embodiment 2 to process tobacco leaves, concrete steps are:
[0040] When treating tobacco leaves, according to the mass ratio of 2% tobacco leaves, spray the above-mentioned bacterial agent evenly on the surface of tobacco leaves with a sprayer, and then adjust the moisture content of tobacco leaves to 25%; after fermenting for a certain period of time under specific temperature and humidity conditions, treat at 80°C for 20 minutes, Inactivates bacteria and the protein enzymes they produce.
[0041] In order to determine the optimum temperature and humidity conditions and time for the action of the bacterial agent, the inventors carried out an orthogonal experiment design, and the related experiment process is briefly introduced as follows.
[0042] Tobacco leaf sample: Take the tobacco leaves after extracting the tobacco stems, divide them into sheets, and then evenly divide them into multiple portions, each portion is 100 g....
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