Metabolite profiling uncovers plasmid-induced cobalt limitation under methylotrophic growth conditions.

PloS One
Patrick KieferJulia A Vorholt

Abstract

The introduction and maintenance of plasmids in cells is often associated with a reduction of growth rate. The reason for this growth reduction is unclear in many cases. We observed a surprisingly large reduction in growth rate of about 50% of Methylobacterium extorquens AM1 during methylotrophic growth in the presence of a plasmid, pCM80 expressing the tetA gene, relative to the wild-type. A less pronounced growth delay during growth under non-methylotrophic growth conditions was observed; this suggested an inhibition of one-carbon metabolism rather than a general growth inhibition or metabolic burden. Metabolome analyses revealed an increase in pool sizes of ethylmalonyl-CoA and methylmalonyl-CoA of more than 6- and 35-fold, respectively, relative to wild type, suggesting a strongly reduced conversion of these central intermediates, which are essential for glyoxylate regeneration in this model methylotroph. Similar results were found for M. extorquens AM1 pCM160 which confers kanamycin resistance. These intermediates of the ethylmalonyl-CoA pathway have in common their conversion by coenzyme B(12)-dependent mutases, which have cobalt as a central ligand. The one-carbon metabolism-related growth delay was restored by providing...Continue Reading

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Citations

Jan 15, 2014·Applied Microbiology and Biotechnology·Frank SonntagJens Schrader
Aug 26, 2015·The ISME Journal·Florian RyffelJulia A Vorholt
Nov 30, 2014·Applied Microbiology and Biotechnology·Andrea M OchsnerJulia A Vorholt
Dec 14, 2017·Proceedings of the National Academy of Sciences of the United States of America·Nina WaleAndrew F Read
Dec 21, 2010·Analytical Chemistry·Patrick KieferJulia A Vorholt

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Methods Mentioned

BETA
PCR

Software Mentioned

Matlab

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