High-yield paenibacillus polymyxa strain and application thereof
A technology of polymyxa spore and bacillus, which is applied in the field of high-yield Paenibacillus polymyxa and its application patent application field, can solve the problems of limited types, limited popularization and application, low fermentation yield and the like
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[0020] The present application will be further explained below in conjunction with the embodiments. Before introducing the specific embodiments, a brief description of some of the experimental backgrounds involved in the following embodiments is as follows.
[0021] Medium:
[0022] Nutrient Agar medium (Nutrient Agar.NA): peptone 10.00 g, beef extract 3.00 g, sodium chloride 5.00 g, agar 15.0 0 g, distilled water 1.00 L, pH 7.0;
[0023] Beef extract medium (Bee4.3 extract Agar): take 500.00 g of fat-free lean meat, chop it, add 1.00 L of distilled water to a beaker, put it in a refrigerator at 4 °C overnight, filter it with cotton, and boil the filtrate in a water bath 1 h, make the protein coagulate, filter, add 10.00 g peptone and 5.00 g sodium chloride to the filtrate, adjust the pH to 7.4, and make up the liquid to 1.00 L, 15.00 g agar, sterilize after aliquoting, and set aside;
[0024] Method for determining the viability of Paenibacillus polymyxa
[0025] Use the d...
Embodiment 1
[0032] In this example, the process of obtaining the Paenibacillus polymyxa YSD7 strain provided by mutagenesis and screening is briefly introduced as follows.
[0033] The Paenibacillus polymyxa YSD7 provided by this application is mainly obtained through ultraviolet mutagenesis and diethyl sulfate (DES) mutagenesis screening of the existing starting strain Paenibacillus polymyxa YSD5, and the specific mutagenesis and screening process are brief described as follows.
[0034] (1) Ultraviolet mutagenesis
[0035] The first is to carry out ultraviolet mutagenesis breeding on the starting strain YSD5, and the main process is as follows:
[0036] Take 4ml of YSD5 bacterial suspension cultured at 30°C to the growth stage of the opponent, and irradiate it with a 15 W ultraviolet lamp at a distance of 28 cm. During the irradiation process, the survival rate of the strain gradually decreases with the extension of the ultraviolet irradiation time. During the radiation mutagenesis pr...
Embodiment 2
[0055] On the basis of the strain obtained in Example 1, the inventors further optimized the fermentation process to further improve the growth and fermentation capacity of the strain. The specific process is introduced as follows.
[0056] (1) Strain activation and seed solution preparation
[0057] After the preserved Paenibacillus polymyxa YSD7 strain was activated and cultured in LB plate medium at 30°C for 2 days, the bacterial lawn was collected and transferred to LB liquid medium for further amplification, and then transferred to the seed medium of the seed tank medium, 30°C, 200rpm fermentation for about 18h as seed liquid;
[0058] The seed medium is: 2% glucose, 3% corn flour, 5% bean cake powder, 1% corn steep liquor powder, 0.2% potassium dihydrogen phosphate, 1.5% yeast powder, pH 7.0;
[0059] (2) Preparation of fermentation broth
[0060] According to the inoculation amount of 6% volume ratio, transfer the seed liquid prepared in step (1) to the fermentation ...
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