Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates

S. Siebenberg, Prashant Madhusudhan Bapat, Anna Eliasson Lantz, B. Gust, L. Heide

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Highly reproducible production values of the aminocoumarin antibiotic novobiocin were achieved by cultivation of a heterologous Streptomyces producer strain in commercially available square deepwell plates consisting of 24 wells of 3 ml culture volume each. Between parallel cultivation batches in the deepwell plates, novobiocin accumulation showed standard deviations of 4-9%, compared to 39% in baffled Erlenmeyer flasks. Mycelia used as inoculum could be frozen in the presence of 20% peptone and stored at 70 degrees C, allowing repeated cultivations from the same batch of inoculum over extended periods of time. Originally, novobiocin titers in the deepwell plate (5-12 mg l(-1)) were lower than in Erlenmeyer flasks (24 mg Optimization of the inoculation procedure as well as addition of a siloxylated ethylene oxide/propylene oxide copolymer, acting as oxygen carrier, to the production medium increased novobiocin production to 54 mg l. The additional overexpression of the pathway-specific positive regulator gene novG increased novobiocin production to 163 mg l(-1). Harvesting the precultures in a defined section of growth phase greatly reduced variability between different batches of inoculum. The use of deepwell plates may considerably reduce the workload and cost of investigations of antibiotic biosynthesis in streptomycetes and other microorganisms due to the high reproducibility and the low requirement for shaker space and culture medium.
    Original languageEnglish
    JournalJournal of Bioscience and Bioengineering
    Volume109
    Issue number3
    Pages (from-to)230-234
    ISSN1389-1723
    DOIs
    Publication statusPublished - 2010

    Keywords

    • Antibiotic production
    • Streptomyces
    • Novobiocin
    • Deepwell-plate
    • Nikkomycin

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