Enhanced ribosome and tRNA contents in Escherichia coli expressing a truncated Vitreoscilla hemoglobin mutant analyzed by flow field-flow fractionation

Abstract

The ribosome and tRNA levels of Escherichia coli cells, transformed with a native or mutated Vitreoscilla hemoglobin genes (vhb), were investigated using asymmetrical flow field-flow fractionation (AFFFF). Mutagenesis of vhb by error-prone PCR was carried out to alter the growth behavior of microaerobically cultivated native VHb-expressing E. coli. A VHb mutant, pVMT1, was identified, which was able to reach a remarkably high final A600 of 15, the value of which being 160% higher than that of a VHb control carrying pVHb8 (A600 5.8). AFFFF revealed that cells expressing mutant vhbs showed up to a doubling in the number of active 70S ribosomes cell−1, an almost 3-fold increase in the number of tRNAs cell−1, and up to a 26% increase in the mass fraction of active 70S ribosomes.

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Correspondence to Leif Bülow.

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Andersson, C.I., Arfvidsson, C., Kallio, P.T. et al. Enhanced ribosome and tRNA contents in Escherichia coli expressing a truncated Vitreoscilla hemoglobin mutant analyzed by flow field-flow fractionation. Biotechnology Letters 25, 1499–1504 (2003). https://doi.org/10.1023/A:1025475703006

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  • asymmetrical flow field-flow fractionation
  • error-prone PCR
  • Escherichia coli
  • oxygen limitation
  • Vitreoscilla hemoglobin (VHb)