Penicillium oxalicum WX-209 strain with high cellulase yield and enzyme producing method
A technology of cellulase and bacterial strains, which is applied in the field of high-yield cellulase strains and enzyme production, and achieves the effect of low equipment requirements, low energy consumption, and improved cellulase activity
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example 1
[0024] Example 1: Screening and identification method of Penicillium oxalicum WX-209
[0025] a. Preliminary screening: samples were collected from 18 places including moldy walnuts in Wenxian County, Gansu Province, soil samples from Yuelu Mountain, orange orchard soil samples, kiwifruit orchard soil samples, and rotten branches from Yuelu Mountain. Take 5 g of the sample and add 100 mL of 2% (W / V) CMC-Na liquid enrichment medium. After enrichment culture at 35°C and 170 r / min shaker for 3 days, take 1 mL of culture solution and dilute to 10 6 -10 8 , take 0.2mL of the diluted solution and spread it evenly on the screening plate medium with CMC-Na as the carbon source. After culturing for 3 days, use the method of Congo red staining to screen out a total of 20 fungal strains with large transparent circles, and then draw a plate The purified strain was finally transferred to PDA slant medium for culture and preservation.
[0026] b. Re-screening: Mandels nutrient solution wa...
Embodiment 2
[0029] Inoculate the spores of Penicillium oxalicum on the slant medium, grow at 35°C for 4 days, wash the spores with sterile water, and make a spore suspension and refrigerate at 4°C for later use.
[0030]Weigh 2.4g of bran and put it into 30ml Mandels nutrient solution for full infiltration. Ultrasonic pretreatment was performed after infiltration, and ultrasonic treatment was performed for 50 minutes under the condition of ultrasonic power 800W. After the treatment, autoclave for 20 minutes, cool to room temperature after sterilization, insert Penicillium oxalicum spores under aseptic conditions, so that the concentration of inoculated spores reaches 1×10 6 individual / mL. The inoculated shake flask was placed in a constant temperature shaker at 35° C. for 72 hours, and the rotation speed was set at 170 r / min. The culture solution was filtered or centrifuged to obtain a crude enzyme solution. The CMCase enzyme activity of the enzyme solution was detected by the DNS metho...
Embodiment 3
[0032] Penicillium oxalicum spores were inoculated on the slant medium, and after growing at 35°C for 3 days, the spores were washed with sterile water to make a spore suspension and refrigerated at 4°C for later use.
[0033] Weigh 2.4g of bran and put it into 30ml Mandels nutrient solution for full infiltration. Ultrasonic pretreatment was performed after infiltration, and ultrasonic treatment was performed for 55 minutes under the condition of ultrasonic power 800W. After the treatment, autoclave for 20 minutes, cool to room temperature after sterilization, insert Penicillium oxalicum spores under aseptic conditions, so that the concentration of inoculated spores reaches 1×10 5 individual / mL. The inoculated shake flask was placed in a constant temperature shaker at 35° C. for 72 hours, and the rotation speed was set at 180 r / min. The culture solution was filtered or centrifuged to obtain a crude enzyme solution. The CMCase enzyme activity of the enzyme solution was detect...
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