Bacillus subtilis culture medium raw material, preparation method and application thereof and culture medium for improving yield of bacillomycin D
A Bacillus subtilis culture medium technology, applied in the field of Bacillus subtilis culture medium for improving the production of bacillus mycin D, can solve the problems of restricting the industrial application of bacillus mycin D and the limited synthesis ability of bacillus mycin D, and achieve the purpose of improving the synthesis Efficiency, the preparation method is simple and easy, and the effect of improving yield
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[0023] The present invention also provides the preparation method of the bacillus subtilis culture medium raw material described in the above technical scheme, comprising the following steps:
[0024] (1) Mix corn stalks, cellulase, xylanase and water, and enzymolyze at 50-60°C for 60-80 hours to obtain an enzymolysis compound;
[0025] (2) Separating the enzymolysis complex from solid to liquid to obtain the enzymolysis supernatant.
[0026] In the present invention, the corn stalks are preferably crushed to a size of 1-2 cm before enzymatic hydrolysis, dried to a constant weight and then crushed to a size above 60 mesh. In the present invention, the drying method is preferably hot-air drying; more preferably, the temperature of the hot-air drying is 60-70°C, more preferably 65°C. The purpose of drying the corn stalks to constant weight in the present invention is to sterilize and inactivate enzymes and facilitate preservation.
[0027] In the present invention, the tempera...
Embodiment 1
[0033] Select mature corn stalks free from diseases and insect pests, remove dust, clean them, cut them into 1-2 cm sections with scissors, put them in an electric blast drying oven, control the temperature at 65°C, and dry them to constant weight. The dried corn stalks are taken out, placed at room temperature for 20 minutes, crushed with a solid high-speed pulverizer, passed through a 60-mesh sieve, and made into powder for later use.
[0034] Accurately weigh 2.00 g of the above-prepared corn stalk powder, put it in a high-pressure steam sterilizer, take it out after sterilizing at 121 ° C for 30 min, cool to room temperature, and add 1 g of cellulase (100,000 U / g) to the sterilized corn stalk powder ), xylanase 2g (100000U / g) and sterile water 54ml, and fully shaken, fully enzymolyzed at a temperature of 50 ° C, the enzymolysis conditions are: cellulase and xylanase addition ratio 2:3 (mass ratio, g:g), enzymolysis time 70h, liquid-material ratio 27mL / g (enzymolysis liquid...
Embodiment 2
[0040] Get respectively 100ml of the enzymolysis supernatants of Example 1, Comparative Example 1 and Comparative Example 2, add yeast extract 2.0g, L-sodium glutamate (MSG) 10g and KH to each enzymolysis supernatant respectively 2 PO4 1.0g, sterilized at 115°C for 20min, cooled to room temperature, prepared bacillusmycin D fermentation medium (Example 1) and control bacillusmycin D fermentation medium 1 (comparative example 1), and control bacillusmycin D fermentation medium Medium 2 (Comparative Example 2).
[0041] Inoculate the Bacillus subtilis preserved in the laboratory on the slant medium, culture at a constant temperature of 37°C for 24 hours, pick a single colony, insert it into the seed medium, and culture it in a shaker flask at 37°C until OD 600 0.8 to 1.0, according to the inoculum size of 6%, respectively inoculated into the above three fermentation media, at 33 ° C, 180 r / min fermentation culture for 110 h.
[0042]After the fermentation, take three fermented ...
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