Clostridium tetani culture medium
A clostridia and culture medium technology, applied in the field of microorganisms, can solve the problems of low toxin production and achieve the effect of increasing the expression amount
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Embodiment 1
[0075] 1. Medium
[0076] Seed medium: soybean peptone total nitrogen 1.60g / L, yeast extract powder 5g / L, potassium dihydrogen phosphate 5g / L, L-cysteine hydrochloride 0.5g / L, resazurin 0.001g / L, Vitamin K1 0.001g / L, agar powder 1.5g / L, pH 7.0±0.1.
[0077] Fermentation medium: soybean peptone total nitrogen 0.90g / L, rice peptone total nitrogen 0.90g / L, yeast extract powder 4.0g / L, sodium acetate 5.0g / L, potassium dihydrogen phosphate 1.00g / L, disodium hydrogen phosphate 1.00g / L, glycerin 60ml / L, glucose 2.5g / L, ferric chloride hexahydrate 0.030, factor Ⅰ 5.0ml / L, factor Ⅱ 2.00ml / L, factor Ⅲ 1.00ml / L, 302 activated carbon 2g / L, pH 7.7±0.1.
[0078] Factor Ⅰ: thiamine hydrochloride 0.30g / L, riboflavin 0.30g / L, pyridoxine hydrochloride 0.30g / L, calcium pantothenate 1.20g / L, biotin 0.004g / L, absolute ethanol 250ml / L.
[0079] Factor II: niacin 2.5g / L, uracil 2.5g / L, L-cystine 75g / L, zinc sulfate heptahydrate 10.0g / L, magnesium sulfate heptahydrate 38.4g / L, concentrated hyd...
Embodiment 2
[0087] Embodiment 2, embodiment 3 strain activation culture medium except that the materials listed in table 1 are different from embodiment 1, all the other components and their content and preparation operation in the strain activation culture medium are all the same as in embodiment 1, and the toxin production culture Basic is the same as embodiment 1, no longer repeats. The results are shown in Table 2.
[0088] The results of the above three examples show that the strain activation medium formulations of Examples 1-Example 3 are all more suitable for the growth of Clostridium tetani, and the strains prepared by the three formulations are inoculated to the toxin-producing medium for fermentation, and the toxin output Correspondingly, it shows that it is more appropriate to add three components of selective soybean peptone, soybean peptone and Hysoy 4D to the strain activation medium.
Embodiment 4- Embodiment 10
[0090] Embodiment 4-embodiment 10 strain activation culture medium is the same as embodiment 3, and toxin-producing culture medium is except that the material listed in table 3 is different, and all the other components and their content and preparation and operation are all the same as embodiment 3, without Let me repeat. The results are shown in Table 4.
[0091] The results of the above 8 examples show that the medium formula of Example 5 is the most suitable formula for Clostridium tetani fermentation, so Example 5 is repeatedly verified, and the following examples 11-13 are repeated verification.
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