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

Paper ID Volume ID Publish Year Pages File Format Full-Text
21458 43223 2010 5 PDF Available
Title
Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates
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 °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.

Keywords
Novobiocin; Nikkomycin; Streptomyces; Deepwell-plate; Antibiotic production
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Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 109, Issue 3, March 2010, Pages 230–234
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us