Application of Iron Ions in Fermentative Production of Low Molecular Weight γ-Polyglutamic Acid by Bacillus subtilis
A Bacillus subtilis, polyglutamic acid technology, applied in fermentation, microorganism-based methods, microorganisms, etc., can solve problems such as the reduction of γ-PGA content, and achieve the effects of improving yield, cheap and efficient production, and broad application prospects.
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Embodiment 1
[0020] Activation of strains: Bacillus subtilis (Bacillus subtilis) PGA-7 strains were inoculated on a solid medium slope, and cultured at 37°C for 16-24 hours to obtain activated Bacillus subtilis strains. The composition of the solid medium: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, agar 20g / L, the balance is water, pH7.0~7.2; the preparation method is to mix the above ingredients After uniformity, adjust the pH value, and then sterilize for later use.
[0021] Preparation of seed solution: Take 2 rings of the above-mentioned activated Bacillus subtilis strains, put them into a 300mL Erlenmeyer flask containing 50mL of fermentation medium, shake and culture at 37°C and 100rpm for 18h to obtain Bacillus subtilis seed solution. The composition of described fermentation medium: citric acid 12g / L, glycerol 80g / L, L-glutamic acid 20g / L, ammonium chloride 7g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate heptahydrate 0.5 g / L, calcium chloride dihydrate ...
Embodiment 2
[0024] The steps of the activation of the strain and the preparation of the seed solution are the same as in Example 1.
[0025] Liquid shake flask fermentation: ferric trichloride is added in the fermentation medium (the formula is the same as the fermentation medium of Example 1), so that the final concentration of the ferric ion is 2.96mmol / L, and it is prepared into a fermentation medium containing iron ions , subpackage 50mL into 300mL Erlenmeyer flask, inoculate the Bacillus subtilis seed liquid into the fermentation medium containing iron ions according to the inoculum amount of 1% by volume fraction, ferment at 37°C, shake and cultivate in a reciprocating shaker for 72h, The rotating speed of the shaking table is 80r / min, and the stroke is 75mm. After the fermentation was completed, the output of gamma-polyglutamic acid was detected to be 25.76g / L. The molecular weight distribution is that the content of γ-polyglutamic acid higher than 245kD accounts for 27.38% of the...
Embodiment 3
[0027] The steps of the activation of the strain and the preparation of the seed solution are the same as in Example 1.
[0028] Liquid shake flask fermentation: ferric chloride is added in the fermentation medium (the formula is the same as the fermentation medium of Example 1), so that the final concentration of the ferric ion is 7.4mmol / L, and it is prepared into a fermentation medium containing iron ions , subpackage 50mL into 300mL Erlenmeyer flask, inoculate the Bacillus subtilis seed liquid into the fermented medium containing ferric ions according to the inoculum amount of 5% volume fraction, ferment at 37°C, shake and cultivate in a reciprocating shaker for 96h, The rotating speed of the shaker is 110r / min, and the stroke is 75mm. After the fermentation was completed, the output of gamma-polyglutamic acid was detected to be 24.69g / L. The molecular weight distribution is that the content of γ-polyglutamic acid higher than 245kD accounts for 9.29% of the total γ-polygl...
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