title: Rapid functional screening of Streptomyces coelicolor regulators by use of a pH indicator and application to the MarR-like regulator AbsC.
abstract: To elucidate the function of an unknown regulator in Streptomyces, differences in phenotype and antibiotic production between a deletion mutant and a wild-type strain (WT) were compared. These differences are easily hidden by complex media. To determine the specific nutrient conditions that reveal such differences, we used a multiwell method containing different nutrients along with bromothymol blue. We found several nutrients that provide key information on characterization conditions. By comparing the growth of wild-type and mutant strains on screened nutrients, we were able to measure growth, organic acid production, and antibiotic production for the elucidation of regulator function. As a result of this method, a member of the MarR-like regulator family, SCO5405 (AbsC), was newly characterized to control pyruvate dehydrogenase in Streptomyces coelicolor. Deletion of SCO5405 increased the pH of the culture broth due to decreased production of organic acids such as pyruvate and alpha-ketoglutarate and increased extracellular actinorhodin (ACT) production in minimal medium containing glucose and alanine (MMGA). This method could therefore be a high-throughput method for the characterization of unknown regulators.
authors.affiliation: School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, Republic of Korea.
authors.forename: Yung-Hun
authors.initials: YH
authors.lastname: Yang
authors.forename: Eunjung
authors.initials: E
authors.lastname: Song
authors.forename: Bo-Rahm
authors.initials: BR
authors.lastname: Lee
authors.forename: Eun-jung
authors.initials: EJ
authors.lastname: Kim
authors.forename: Sung-Hee
authors.initials: SH
authors.lastname: Park
authors.forename: Yun-Gon
authors.initials: YG
authors.lastname: Kim
authors.forename: Chang-Soo
authors.initials: CS
authors.lastname: Lee
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001431:Bacteriological Techniques / Q000379:methods*; D002264:Carboxylic Acids / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry*; D017353:Gene Deletion; D057166:High-Throughput Screening Assays; D006863:Hydrogen-Ion Concentration; D048372:Streptomyces coelicolor / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002264:Carboxylic Acids; D003470:Culture Media; D014157:Transcription Factors
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