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Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens

Delaney, Nigel F ; Kaczmarek, Maria E ; Ward, Lewis M ; Swanson, Paige K ; Lee, Ming-Chun ; Marx, Christopher J Zhang, Qijing

PloS one, 2013-04, Vol.8 (4), p.e62957-e62957 [Periódico revisado por pares]

United States: Public Library of Science

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Título:
Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens
Autor: Delaney, Nigel F ; Kaczmarek, Maria E ; Ward, Lewis M ; Swanson, Paige K ; Lee, Ming-Chun ; Marx, Christopher J
Zhang, Qijing
Assuntos: Analysis ; Automation ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Batch culture ; Biofilms ; Biology ; Carbon - metabolism ; Carbon compounds ; Cell culture ; Cellulose ; Cellulose synthase ; Chelating Agents - pharmacology ; Citric acid ; Cobalt ; Culture media ; Culture Media - chemistry ; Dehydrogenases ; Ethylenediaminetetraacetic acids ; Evolutionary biology ; Genomes ; Growth media ; Growth rate ; Handbooks ; Laboratories ; Laboratory equipment ; Metabolism ; Metals ; Methods ; Methylobacterium extorquens ; Methylobacterium extorquens - drug effects ; Methylobacterium extorquens - genetics ; Methylobacterium extorquens - growth & development ; Methylobacterium extorquens - metabolism ; Mutation ; Physiological aspects ; Pipes ; Prokaryotes ; Public software ; Response surface methodology ; Robotics ; Strains (organisms)
É parte de: PloS one, 2013-04, Vol.8 (4), p.e62957-e62957
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
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Conceived and designed the experiments: NFD MEK LMW PKS ML CJM. Performed the experiments: NFD MEK LMW PKS ML. Analyzed the data: NFD MEK LMW PKS ML CJM. Contributed reagents/materials/analysis tools: NFD LMW. Wrote the paper: NFD CJM.
Competing Interests: The authors have declared that no competing interests exist.
Current address: Division of Geological and Planetary Sciences, California Institute of Technology, Pasadena, California, United States of America
Current address: Department of Biochemistry, The University of Hong Kong, Pokfulam, Hong Kong
Current address: Department of Molecular Genetics and Microbiology, University of Texas, Austin, Texas, United States of America
Descrição: Methylobacterium extorquens strains are the best-studied methylotrophic model system, and their metabolism of single carbon compounds has been studied for over 50 years. Here we develop a new system for high-throughput batch culture of M. extorquens in microtiter plates by jointly optimizing the properties of the organism, the growth media and the culturing system. After removing cellulose synthase genes in M. extorquens strains AM1 and PA1 to prevent biofilm formation, we found that currently available lab automation equipment, integrated and managed by open source software, makes possible reliable estimates of the exponential growth rate. Using this system, we developed an optimized growth medium for M. extorquens using response surface methodologies. We found that media that used EDTA as a metal chelator inhibited growth and led to inconsistent culture conditions. In contrast, the new medium we developed with a PIPES buffer and metals chelated by citrate allowed for fast and more consistent growth rates. This new Methylobacterium PIPES ('MP') medium was also robust to large deviations in its component ingredients which avoided batch effects from experiments that used media prepared at different times. MP medium allows for faster and more consistent growth than other media used for M. extorquens.
Editor: United States: Public Library of Science
Idioma: Inglês

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Development of an optimized medium, strain and high-throughput culturing methods for Methylobacterium extorquens

Delaney, Nigel F ; Kaczmarek, Maria E ; Ward, Lewis M ; Swanson, Paige K ; Lee, Ming-Chun ; Marx, Christopher J Zhang, Qijing

United States: Public Library of Science

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