Antimicrobial Peptide Extraction

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Purpose

This protocol is used for extracting antimicrobial peptides made by bacterial species.

Materials Needed

Occidiofungin extraction

-Potato dextrose broth

-Burkholderia contaminans

-large centrifuge tubes

-centrifuge

-35% Acetonitrile

-2000 mL flasks

-stir bar

-stir plate

-Sodium Thiosulfate

-plastic pipette

Protocol

Occidiofungin extraction

-Culture the Burkholderia contaminans bacteria in Potato Dextrose Broth at 25 C for at least 5 days

-Centrifuge the bacteria at 13,000 rpm for 20 minutes

-Pour the supernatant into 2000 mL glass flask with a stir bar and add 0.5 volume of Sodium Thiosulfate (i.e. 750 mL of supernatant, 375 grams Sodium Thiosulfate)

-Stir the mixture for 2 hours

-Centrifuge the mixture at 13,000 rpm for 20 minutes

-Discard the supernatant and let the protein precipitate dry overnight

-Wash the protein out of the centrifuge tubes with 35% Acetonitrile and a plastic pipet into a plastic tube

References

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