Global phenotypic characterization of bacteria

Bochner, Barry R.

doi:10.1111/j.1574-6976.2008.00149.x

Cited by 363 publications

(335 citation statements)

References 86 publications

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“…Preliminary results from Biolog-AN plates (data not shown) indicated that not only uptake and oxidation of amino acids, but also the ability to transport and oxidize many other compounds was reduced early during irradiation. The ability to reduce substrates in the assay was lost simultaneously for all C-sources, again indicating that the final step of the respiratory chain is inactivated at this time rather than single transport systems within the membrane (Bochner, 2009). …”

Section: Discussion Changes In Cellular Energy Metabolism During Uva mentioning

confidence: 99%

The respiratory chain is the cell's Achilles' heel during UVA inactivation in Escherichia coli

et al. 2010

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Solar disinfection (SODIS) is used as an effective and inexpensive tool to improve the microbiological quality of drinking water in developing countries where no other means are available. Solar UVA light is the agent that inactivates bacteria during the treatment. Damage to bacterial membranes plays a crucial role in the inactivation process. This study showed that even slightly irradiated cells (after less than 1 h of simulated sunlight) were strongly affected in their ability to maintain essential parts of their energy metabolism, in particular of the respiratory chain (activities of NADH oxidase, succinate oxidase and lactate oxidase were measured). The cells' potential to generate ATP was also strongly inhibited. Many essential enzymes of carbon metabolism (glucose-6-phosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase and malate dehydrogenase) and defence against oxidative stress (catalases and glutathione-disulfide reductase) were reduced in their activity during SODIS. The work suggests that damage to membrane enzymes is a likely cause of membrane dysfunction (loss of membrane potential and increased membrane permeability) during UVA irradiation. In this study, the first targets on the way to cell death were found to be the respiratory chain and F 1 F 0 ATPase.

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Section: Discussion Changes In Cellular Energy Metabolism During Uva mentioning

confidence: 99%

The respiratory chain is the cell's Achilles' heel during UVA inactivation in Escherichia coli

et al. 2010

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“…In total, these 10 PM plates contained 237 toxins (including antibacterial agents), with each toxin present at four concentrations (24). Growth in each toxin-containing well could be monitored by purple color development because of the presence of a tetrazolium indicator dye (25). A negative control (E. coli harboring the empty ASKA vector, pCA24N-NoIns) was used to inoculate a second set of PM plates in parallel.…”

Section: Resultsmentioning

confidence: 99%

Artificial gene amplification reveals an abundance of promiscuous resistance determinants in Escherichia coli

Soo

Hanson‐Manful²,

Patrick³

2010

Proc. Natl. Acad. Sci. U.S.A.

111

110

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Duplicated genes provide an important raw material for adaptive evolution. However, the relationship between gene duplication and the emergence of new biochemical functions is complicated, and it has been difficult to quantify the likelihood of evolving novelty in any systematic manner. Here, we describe a comprehensive search for artificially amplified genes that are able to impart new phenotypes on Escherichia coli, provided their expression is up-regulated. We used a high-throughput, library-on-library strategy to screen for resistance to antibiotics and toxins. Cells containing a complete E. coli ORF library were exposed to 237 toxin-containing environments. From 86 of these environments, we identified a total of 115 cases where overexpressed ORFs imparted improved growth. Of the overexpressed ORFs that we tested, most conferred small but reproducible increases in minimum inhibitory concentration (≤16-fold) for their corresponding antibiotics. In many cases, proteins were acting promiscuously to impart resistance. In the absence of toxins, most strains bore no fitness cost associated with ORF overexpression. Our results show that even the genome of a nonpathogenic bacterium harbors a substantial reservoir of resistance genes, which can be readily accessed through overexpression mutations. During the growth of a population under selection, these mutations are most likely to be gene amplifications. Therefore, our work provides validation and biochemical insight into the innovation, amplification, and divergence model of gene evolution under continuous selection [Bergthorsson U, Andersson DI, Roth JR (2007) Proc Natl Acad Sci USA 104:17004-17009], and also illustrates the high frequency at which novel traits can evolve in bacterial populations.antibiotic resistance | evolutionary innovation | molecular evolution | protein promiscuity | phenotype microarray

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“…Bacteria grown on solid medium are suspended in a minimal, defined medium (Bochner, Gadzinski and Panomitros, 2001;Bochner, 2009). The phenotypic microarray is carried out in 96-well plates, with each of the wells containing a different nutrient.…”

Section: Phenotypic Microarray Technologymentioning

confidence: 99%