Aureobasidin A high-yield bacterial strain and application thereof
A technology for bacterial strains and bacterial suspensions, applied in fungi, biochemical equipment and methods, microorganisms, etc., can solve the problems of increasing the yield of AureobasidinA, which can reduce large-scale production costs, improve production capacity, and have strong antifungal activity. Effect
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Embodiment 1
[0050] Example 1: Acquisition of AureobasidinA Original Producer Bacteria
[0051] Soil samples were taken from the lower layer of decomposed leaves in Moganshan Scenic Area, Hangzhou City, Zhejiang Province. The sampling volume was 5g, diluted with sterile water and evenly spread to the PDA with 50 μg / ml ampicillin sodium and 50 μg / ml streptomycin sulfate added at the same time. On the culture medium, culture them upside down in a constant temperature and humidity chamber at 25° C. and 60% relative humidity for 3 days. Take a flat plate with independent single colony distribution, and use a sterile toothpick 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. Number and number the colonies that are creamy white in the early stage but turn black...
Embodiment 2
[0056] Example 2: Confirmation of the ability of A. pullulans HDCC101-R13 strain to produce AureobasidinA
[0057] The isolated and purified A. pullulans HDCC101-R13 strain was inoculated on PDA solid medium, and cultured upside down in a constant temperature and humidity chamber at 25°C and 60% relative humidity for 7 days. Take an appropriate amount of bacterial lawn with an inoculation loop, inoculate it in 100ml sterile liquid seed medium, and culture it on a shaker at 25°C and 220rpm for 48h. After microscopic examination of no bacteria, use a pipette to inoculate 0.4 ml of the seed solution into a 250 ml Erlenmeyer flask containing 40 ml of fermentation medium, inoculate 5 bottles of fermentation medium in total, and place it at 25° C. for 8 days on a shaker at 220 rpm. After the fermentation and cultivation, take 4ml of the fermentation broth, mix it with 1 times the volume of absolute ethanol, soak it in ultrasonic for 20min, mix it again and centrifuge it at 3000rpm f...
Embodiment 3
[0058] Example 3: Preparation and verification of AureobasidinA high-yielding strains
[0059] 1) HDCC101-R13 was used as the original starting strain, inoculated into PDA solid medium, and cultured 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 plate of grown solid cultured bacterial lawn, scrape off the bacterial lawn with a sterile steel swab, transfer it to a triangular flask with glass beads and 15ml sterile saline, wrap it up and place it on a shaker to shake and disperse After 30 minutes, a well-dispersed bacterial suspension was obtained.
[0061] 3) After taking the prepared bacterial suspension for mutagenesis treatment, evenly spread it on the PDA solid medium added with a specific resistance reagent, and culture it upside down in a constant temperature and humidity chamber at 25°C and 60% relative humidity for 4 days, and then Spot them on fresh PDA solid medium...
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