Antifungal compound generated by marine aspergillus and preparation method thereof
A kind of compound, mycin technology, applied in the field of medicine and biology
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Embodiment 1
[0067] identification of bacterial strain IMB17-055 (Aspergillus versicolor)
[0068] a) Source of the strain: The strain Aspergillus versicolor IMB17-055 (Aspergillus versicolor) was isolated from the sea mud samples (18°24′09.00″N, 109°51′08.00″E) collected from the Sanya Hongsha mangrove forest.
[0069] b) Identification of strains: According to the conservation of the DNA sequence in the ITS region among microbial species, the fungus IMB17-055 (Aspergillus versicolor) was identified. The genome of the strain was extracted, its ITS gene was amplified by PCR and sequenced, and submitted to the NCBI GenBank database for accession number MN294468.
[0070] SEQ ID NO.1
[0071] >IMB17-055 ITS
[0072] 1 agtcgttacg tagtgacctg cggaaggatc attackgagt gcgggctgcc tccgggcgcc
[0073] 61 caacctccca cccgtgacta cctaacactg ttgcttcggc ggggagccct ctcgggggcg
[0074] 121 agccgccggg gactactgaa cttcatgcct gagagtgatg cagtctgagt ctgaatataa
[0075] 181 aatcagtcaa aactttcaac aatggatctc ttg...
Embodiment 2
[0083] Co-culture screening of Aspergillus versicolor (Aspergillus versicolor) IMB17-055
[0084]The strain Aspergillus versicolor IMB17-055 and other co-culture strains, including Acremonium persicinum IMB17-217, Aspergillus oryzae IMB17-008, Aspergillussp.IMB17-028, Aspergillus sp.IMB17-036, Aspergillus sp.IMB17 -078, Aspergillus jensenii IMB17-079, Aspergillus chevalieri IMB18-208, Cladosporium xanthochromaticum IMB17-002, Penicillium raistrickii IMB17-035, Penicillium glabrum IMB18-209, Penicillium chrysogenum IMB176 etc. Inoculate on a PDA medium plate [recipe: 2.0 g of glucose, 0.3 g of potato extract powder, 18 g of agar powder, 100 mL of deionized water], and culture in a constant temperature incubator at 28°C for 7-14 days. Take about 1cm with a sterile shovel 2 The bacteria-containing agar block was inoculated in 20mL liquid PDB fermentation medium [formulation: glucose 2.0g, potato extract powder 0.3g, deionized water 100mL], in a 100mL Erlenmeyer flask, 28°C, 200...
Embodiment 3
[0088] Co-cultivation of Aspergillus versicolor (Aspergillus versicolor) IMB17-055 prepared by mass fermentation
[0089] Inoculate the spore liquid of strain IMB17-055 on a PDA medium plate [recipe: glucose 2.0g, potato extract powder 0.3g, agar powder 18g, deionized water 100mL], and place it in a constant temperature incubator at 28°C for 7- 14 days, take about 1cm with a sterile shovel 2 The bacteria-containing agar block was inoculated into 100mL liquid PDB fermentation medium, and cultured in a 500mL Erlenmeyer flask at 28°C and 200r / min for 3 days in a shaker flask to prepare a seed solution.
[0090] Using the same procedure, a seed liquid of strain Aspergillus chevalieri IMB18-208 was prepared. The seed liquids of the two strains were mixed in a ratio of 1:1, and then the mixed seed liquid was transferred to the PDB fermentation medium at an inoculum size of 10%, and a total of 30 L was cultured at 28 ° C and 200 rpm for 10 days with shaking.
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