Penicillium oxalicum bam-1 and its separation and purification method and application
A Penicillium oxalate, application method technology, applied in the field of microorganisms, can solve the problems of no ecology, environmental protection and the like
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Embodiment 1
[0027] Embodiment 1: Isolation, activation and cultivation of bacterial strains
[0028] The strain was isolated from the organic mulch and the soil covering the Phyllostachys praecox forest that had been piled up for a long time in Gaoyun Village, Taihuyuan Town, Lin'an City, Zhejiang Province. It was a pure strain obtained through enrichment culture of samples, primary screening, and secondary screening.
[0029] Enrichment culture of samples: Weigh 10.0 g of soil and decayed organic mulch samples, add them into a 250 ml Erlenmeyer flask filled with 90 ml of sterile water and glass beads, 180 r min -1 Shake for 30 min to fully disperse the sample, use a sterile pipette to take 5 ml of the suspension and inoculate it into 45 ml of enrichment medium, at 28 °C, 120 r min -1 Cultured with shaking for 2 days. Then take the culture solution and inoculate it into fresh sterile enrichment medium, and culture repeatedly for 3 times.
[0030] The enrichment medium adopts the impro...
Embodiment 2
[0039] Example 2: Strain filter paper enzyme activity
[0040] Penicillium oxalicum ( Penicillium oxalicum) BAM-1, Trichoderma viride, and Trichoderma harzianum were inoculated in Hutchison liquid medium (formula: 1 cm×6 cm Xinhua quantitative filter paper, KH 2 PO 4 1.05g, NaCl 0.11g, MgSO 4 ·7H 2 O 0.29 g, NaNO 3 2.51 g, FeCl 3 0.01g, CaCl 2 0.10g, 1000 ml water, pH 6.05), 120 r·min at 28°C -1 Shake flask shaking culture, see the results figure 1 , it was found that the filter paper enzyme activities of the three strains first increased and then decreased with the prolongation of culture time, and the enzyme activities were the highest at 5 days of culture, which were 9.83 U, 3.93 U, and 4.64 U, respectively, and Penicillium oxalicum ( Penicillium oxalicum) The enzyme activity of BAM-1 filter paper is more than 2 times that of Trichoderma viride and Trichoderma harzianum.
Embodiment 3
[0041] Example 3: Strain Enzyme Production Medium Optimization
[0042] Based on the single factor test, the L 9 (3 4 ) Orthogonal experimental design, for carbon sources containing different concentrations (rice straw 10 g·L -1 , 15 g·L -1 , 25g L -1 ), nitrogen source (beef extract 1.5 g L -1 , 2.0 g L -1 , 2.5 g L -1 ) and inorganic salts (FeCl 3 0.01 g·L -1 , 0.015 g·L -1 , 0.02 g g L -1 and KH 2 PO 4 1.0g L -1 , 1.5 g·L -1 , 2.0 g L -1 ) Hutchinson liquid medium was optimized, see Table 1. The optimal combination of medium factors for enzyme production by strains is: rice straw 25 g·L -1 , beef extract 2.5 g·L -1 , FeCl 3 0.01 g·L -1 、KH 2 PO 4 1.5 g·L -1 .
[0043] Table 1L 9 (34 ) Orthogonal Experimental Design and Results
[0044]
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