| Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates. | |
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MedLine Citation:
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PMID: 20159569 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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 l(-1)). 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(-1). 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. |
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Authors:
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Stefanie Siebenberg; Prashant M Bapat; Anna Eliasson Lantz; Bertolt Gust; Lutz Heide |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't Date: 2009-09-16 |
Journal Detail:
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Title: Journal of bioscience and bioengineering Volume: 109 ISSN: 1347-4421 ISO Abbreviation: J. Biosci. Bioeng. Publication Date: 2010 Mar |
Date Detail:
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Created Date: 2010-02-17 Completed Date: 2010-05-18 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 100888800 Medline TA: J Biosci Bioeng Country: Japan |
Other Details:
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Languages: eng Pagination: 230-4 Citation Subset: IM |
Copyright Information:
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Copyright 2009 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved. |
Affiliation:
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Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhardt Karls-Universit?t T?bingen, Auf der Morgenstelle 8, 72076 T?bingen, Germany. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Anti-Bacterial Agents
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biosynthesis* Bioreactors / microbiology* Cell Culture Techniques / instrumentation* Equipment Design Equipment Failure Analysis Novobiocin / biosynthesis* Streptomyces / cytology, metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Anti-Bacterial Agents; 303-81-1/Novobiocin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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