Building an Aquatic Actinomycete Library: Identification and Secondary Metabolite Profiling
Poster Number
20C
Research or Creativity Area
Natural Sciences
Abstract
Actinomycetes are a group of Gram-positive bacteria that dedicate a large portion of their genome to biosynthetic machinery that constructs secondary metabolites. In the environment these metabolites are used for various functions, however these small molecules often have beneficial bioactivities for humans: antibiotic, antifungal, anticancer, antiviral, etc. Actinomycetes have been found in both terrestrial and marine habitats however marine actinomycetes have been found to be genetically distinct from terrestrial species. Freshwater actinomycetes are an under-explored source of these Gram-positive bacteria with few studies published within the US and even fewer from freshwater sponges. In an era where antibiotic resistance is becoming more prevalent, it is more important now than ever to continue the pursuit of new drug leads. The Carlson Lab has been hard at work isolating novel actinomycete strains from aquatic environments. These isolates are grown up in 2 L scale for 4-7 d and extracted using XAD-16 resin beads. The chemistry extracted by the resin is released by rinsing the beads with acetone. The acetone is then dried down and partitioned with ethyl acetate. The crude extract is then fractionated using silica gel chromatography solid phase extraction columns. These fractions are then analyzed for chemical complexity using high-performance liquid chromatography and liquid chromatography mass spectrometry. The fractions are also tested for antibiotic activity against four human pathogens: Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Salmonella sp.. Genetic identification via 16s rRNA sequencing is currently underway. The creation of the microbial and chemical library will be discussed.
Location
Don and Karen DeRosa University Center (DUC) Poster Hall
Start Date
27-4-2024 10:30 AM
End Date
27-4-2024 12:30 PM
Building an Aquatic Actinomycete Library: Identification and Secondary Metabolite Profiling
Don and Karen DeRosa University Center (DUC) Poster Hall
Actinomycetes are a group of Gram-positive bacteria that dedicate a large portion of their genome to biosynthetic machinery that constructs secondary metabolites. In the environment these metabolites are used for various functions, however these small molecules often have beneficial bioactivities for humans: antibiotic, antifungal, anticancer, antiviral, etc. Actinomycetes have been found in both terrestrial and marine habitats however marine actinomycetes have been found to be genetically distinct from terrestrial species. Freshwater actinomycetes are an under-explored source of these Gram-positive bacteria with few studies published within the US and even fewer from freshwater sponges. In an era where antibiotic resistance is becoming more prevalent, it is more important now than ever to continue the pursuit of new drug leads. The Carlson Lab has been hard at work isolating novel actinomycete strains from aquatic environments. These isolates are grown up in 2 L scale for 4-7 d and extracted using XAD-16 resin beads. The chemistry extracted by the resin is released by rinsing the beads with acetone. The acetone is then dried down and partitioned with ethyl acetate. The crude extract is then fractionated using silica gel chromatography solid phase extraction columns. These fractions are then analyzed for chemical complexity using high-performance liquid chromatography and liquid chromatography mass spectrometry. The fractions are also tested for antibiotic activity against four human pathogens: Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Salmonella sp.. Genetic identification via 16s rRNA sequencing is currently underway. The creation of the microbial and chemical library will be discussed.
