Computational protocol: New Spirotetronate Antibiotics, Lobophorins H and I, from a South China Sea-Derived Streptomyces sp. 12A35

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Protocol publication

[…] The sediment was collected from the South China Sea at the depth of 2134 m (17°59.928′N, 111°36.160′E). To effective isolation of culturable marine actinomycetes by making the culture conditions similar to true marine environment, the strain 12A35 was isolated after incubation at 28 °C for 1 week on modified Gauze’s synthetic medium NO. 1 with artificial sea water instead of NaCl and distilled water (soluble starch 20.0 g; KNO3 1.0 g; MgSO4·7H2O 0.5 g; K2HPO4 0.5 g; FeSO4·7H2O 10.0 mg; agar 15.0 g; artificial sea water 1.0 L; adjust pH 7.0). Genomic DNA isolation, PCR amplification of 16S rDNA, and sequencing were performed with conventional methods. Sequence analysis of 16S rDNA were performed using BLASTN. Phylogenetic tree were constructed using Neighbor-Joining method of MEGA (version 5.0). Tree topologies were evaluated by bootstrap analysis with 1000 replicates. The strain 12A35 was maintained on modified Gauze’s synthetic medium NO. 1 at 28 °C, and the agar was cut into pieces (1 × 1 cm) and inoculated into 5 × 100 mL of seed medium (potato sucrose broth containing sucrose 20 g, infusion from 200 g potatoes, artificial sea water 1 L at pH 6.0 before sterilization) in 500-mL Erlenmeyer flasks, then cultivated at 28 °C for 2 days with shaking at 180 rpm. Seed cultures were transferred into 24 × 650 mL production medium in 3.0 L Erlenmeyer flasks, with an inoculation volume of 3%–4% (v/v), and incubated under the same conditions for 7 days. […]

Pipeline specifications

Software tools BLASTN, MEGA
Application Phylogenetics
Organisms Bacillus subtilis, Staphylococcus aureus
Diseases Multiple Sclerosis
Chemicals Ampicillin, Silica Gel