A kind of aureobasidin A high-yield bacterial strain and its application
A technology of strains and microbial strains, applied in the field of high-yielding AureobasidinA strains, can solve the problems of increasing the yield of AureobasidinA, which has not been seen, and achieve the effects of reducing large-scale production costs, strong antifungal activity, and improving production capacity
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Embodiment 1
[0050] Example 1: Acquisition of the original producing bacteria of AureobasidinA
[0051] Soil samples were taken from the lower layer of decayed leaves in Moganshan Scenic Area, Hangzhou City, Zhejiang Province, with a sampling amount of 5g, which was diluted with sterile water and then evenly spread on PDA supplemented with 50μg / ml ampicillin sodium and 50μg / ml streptomycin sulfate. On the medium, the cells were cultured upside down in a constant temperature and humidity chamber at 25°C and 60% relative humidity for 3 days. Take the plate with independent single colony distribution, and use sterile toothpicks to inoculate the creamy colonies on fresh PDA medium supplemented with 80 μg / ml ampicillin sodium and 80 μg / ml streptomycin sulfate, at 25 ° C, 60% The cells were cultured upside down in a constant temperature and humidity chamber with relative humidity for 9 days and observed every day. The colonies that were creamy white in the early stage but turned black in the la...
Embodiment 2
[0056] Example 2: Confirmation of AureobasidinA-producing ability of A. pullulans HDCC101-R13 strain
[0057] The isolated and purified strain of A. pullulans HDCC101-R13 was inoculated into PDA solid medium, and cultured upside down in a constant temperature and humidity chamber at 25°C and 60% relative humidity for 7 days. An appropriate amount of bacterial moss was taken with an inoculation loop, inoculated into 100 ml of sterile liquid seed medium, and cultured on a shaker at 25°C and 220 rpm for 48 hours. After microscopic examination, 0.4ml of seed liquid was inoculated into a 250ml conical flask containing 40ml of fermentation medium, and 5 bottles of fermentation medium were inoculated, and cultured on a shaker at 25°C and 220rpm for 8 days. After the fermentation culture, take 4 ml of the fermentation broth, mix it with 1 volume of absolute ethanol, soak it in ultrasonic for 20 minutes, and then mix it again and centrifuge it at 3000 rpm for 15 minutes. The supernatan...
Embodiment 3
[0058] Example 3: Preparation and verification of AureobasidinA high-yielding strains
[0059] 1) Take HDCC101-R13 as the original starting strain, inoculate it into PDA solid medium, and cultivate it upside down in a constant temperature and humidity chamber at 25°C and 60% relative humidity for 5 days to obtain fresh provenance.
[0060] 2) Take 1 dish of the grown solid culture bacterial moss, scrape off the bacterial moss with a sterile steel stick, transfer it to a triangular flask with glass beads and 15ml sterile saline, wrap it and place it on a shaker to shake and break up 30min to obtain a well-dispersed bacterial suspension.
[0061] 3) Take the prepared bacterial suspension and evenly coat it on the PDA solid medium with specific resistance reagent after mutagenesis treatment, and invert the culture in a constant temperature and humidity chamber at 25°C and 60% relative humidity for 4 days, then They were seeded on fresh PDA solid medium supplemented with specific...
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