A spinosad fermentation method for improving cell activity
A spinosyn and fermentation method technology, which is applied in the field of biological pesticide preparation, can solve the problems of unsatisfactory spinosyn fermentation yield and other problems, and achieve the effects of obvious economic benefit and value, increase fermentation yield, and improve cell activity.
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Embodiment 1
[0028] A spinosad fermentation method for improving cell activity, comprising steps as follows:
[0029] (1) After the experimental verification of the strains producing spinosad, the glycerin tube passed through the seed bottle and fermented for 2-3 days. Microscopic examination showed no bacteria. The shape of the bacteria was radial without breaks. It is 6.8, grow to maturity, obtain seed liquid; carry out next step fermentation.
[0030] (2) Add seed medium into a 30-liter seed tank, cool to 30°C after sterilization, and carry out aseptic pressure holding, and then under the protection of the flame, put the seed liquid into the seed tank for seed cultivation, the inoculation amount 50ml, the seed tank culture conditions are: temperature 30°C, tank pressure 0.05Mp, ventilation rate 1vvm, pH between 7-8, culture for 2-3 days, stirring speed 300 rpm, dissolved oxygen general seed tank maintenance Above 80%, the raw material formula of the seed medium is composed as follows i...
Embodiment 2
[0035] A spinosad fermentation method for improving cell activity, comprising steps as follows:
[0036] (1) After the experimental verification of the strains producing spinosad, the glycerol tube passed through the seed bottle and fermented for 2-3 days. Microscopic examination showed no bacteria, the shape of the bacteria was radial, no breaks, the bacterial concentration reached more than 20%, and the pH It is 6.8, grow to maturity, obtain seed liquid; carry out next step fermentation.
[0037] (2) Add seed culture medium into a 3000-liter seed tank, cool to 30°C after sterilization, and carry out aseptic pressure holding, and then under the protection of flame, put the seed liquid into the seed tank for seed cultivation, the inoculation amount The seed tank culture conditions are: temperature 30°C, tank pressure 0.05Mp, ventilation volume 1vvm, pH between 7-8, culture for 2-3 days, the stirring speed is 100 rpm for the first 0-10 hours, and then the stirring speed For 25...
experiment example
[0043] Ten batches of fermenters were selected, and zinc sulfate and cobalt chloride were added to the basic material, and the cell activity and potency of spinosad were measured at various time periods during the fermentation of spinosyn fermenters.
[0044] Cell viability assay method: MTT staining method was used to measure absorbance with a microplate reader to measure cell viability.
[0045] Potency test method: high performance liquid chromatography, according to the method specified in the national standard GB29696-2017.
[0046] In summary, the addition of metal trace elements zinc sulfate and cobalt chloride to spinosyn fermentation can significantly improve cell activity, and also significantly increase the yield of spinosad. Plays an important role in fermentation production.
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