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Developing An Electroporation Method For Transforming Streptomyces Nymphaeiformis, Heather Knott, Stephen Baron Apr 2022

Developing An Electroporation Method For Transforming Streptomyces Nymphaeiformis, Heather Knott, Stephen Baron

Honors Projects

Streptomyces species are notoriously difficult to transform. Streptomyces nymphaeiformis is no different, so a method of electroporation was used to attempt to transform the cells. Multiple growth stages were used in order to alter the degree of development of the cell wall. The procedure did not kill the cells, but the cells were not transformed. Due to the lack of transformation with S. nymphaeiformis, transformation was attempted on two other Streptomyces strains, S. lividans and S. coelicolor. Neither was successfully transformed to thiostrepton (tsr) resistance, nor did they grow on a plate lacking thiostrepton. One possibility for the …


Characterization Of Streptomyces Nymphaeiformis Sp. Nov., And Its Taxonomic Relatedness To Other Polyhydroxybutyrate-Degrading Streptomycetes, Gary R. Hix, Muhammad S. Khan, Mikayla T. Miller, Elisha C. Napier, Allison L. O'Brien, Roger P. White, Stephen F. Baron Ph.D. Mar 2022

Characterization Of Streptomyces Nymphaeiformis Sp. Nov., And Its Taxonomic Relatedness To Other Polyhydroxybutyrate-Degrading Streptomycetes, Gary R. Hix, Muhammad S. Khan, Mikayla T. Miller, Elisha C. Napier, Allison L. O'Brien, Roger P. White, Stephen F. Baron Ph.D.

Biology Faculty Scholarship

A polyhydroxybutyrate (PHB)-degrading actinomycete, strain SFB5AT, was identified as a species of Streptomyces based on its membrane fatty acid profile and the presence of LL-diaminopimelic acid in the cell wall. It formed sporulating mycelia on most agar media, but flat or wrinkled, moist colonies on trypticase soy agar. Spores were smooth, cylindrical, and borne on long, straight to flexuous chains. It produced a light brown diffusible pigment, but not melanin. Comparison of genomic digital DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) values indicated that strain SFB5AT was related to Streptomyces litmocidini JCM 4394T , Streptomyces …


Possible Role Of N-Acetylglucosamine For Induction Of Polyhydroxybutyrate Depolymerase In Streptomyces Sp. Sfb5a, Morgan Todd May 2020

Possible Role Of N-Acetylglucosamine For Induction Of Polyhydroxybutyrate Depolymerase In Streptomyces Sp. Sfb5a, Morgan Todd

Honors Projects

Streptomyces sp. SFB5A is a filamentous, Gram-positive bacterium that makes a polyhydroxybutyrate (PHB) depolymerase to degrade PHB for energy usage. Its life cycle begins as spores germinate to form vegetative mycelia. Starvation prompts formation of aerial mycelia and ultimately spores after 5-7 days. During starvation, Streptomyces may use peptidoglycan hydrolases to degrade its own cell walls, releasing N-Acetylglucosamine (NAG). PHA depolymerase activity is detected during growth of strain SFB5A on NAG as the sole carbon source, suggesting that NAG induces PHA depolymerase synthesis. To test this hypothesis, strain SFB5A was grown for 4 days on PHB, and a colorimetric assay …


Glycogen Accumulation By Wild Type And Bald Mutant Strains Of Streptomyces Sp. Sfb5a During Growth On 3-Hydroxybutyrate, Mackenzie Francis May 2020

Glycogen Accumulation By Wild Type And Bald Mutant Strains Of Streptomyces Sp. Sfb5a During Growth On 3-Hydroxybutyrate, Mackenzie Francis

Honors Projects

Previous research has shown that Streptomyces sp. SFB5A produces PHA depolymerase and forms aerial hyphae, while the bld4 mutant of this species forms aerial hyphae very poorly and does not produce PHA depolymerase. This effect may be due to the mutant’s inability to sense starvation. Therefore, we hypothesized that the bld4 may also be deficient in formation of glycogen and spores, which are both associated with starvation. To test this hypothesis, we grew the wild type (WT) Streptomyces sp. SFB5A and bld4 in broth cultures containing 3HB. We compared 3HB consumption, glycogen accumulation, PHA depolymerase synthesis, protein accumulation, and spore …