Aldehyde-reactive probe -based

Fig. 2. Quantitation of DNA damage. Borrelia burgdorferi B31A cells were grown in BSK-II under anaerobic conditions to a cell density of 5 x 10 cells/mL, treated with 0, 10, or 50 mM f-butyl peroxide (C, D, and E, respectively) for 4 h and DNA isolated. The DNA was then mixed with an aldehyde-reactive probe (Oxford Biomedical Research, Inc) labeled with biotin and detected with an HRP-streptavidin conjugate. The color development was monitored at 450 nm. The number of aldehyde-reactive probe (DNA base lesions)/10 bp DNA was determined using a standard curve. Escherichia coli TA4315 cells were grown in minimal media to ODgoo of 0.4, treated with 0 or 100 mM H2O2 for 30 min (A and B, respectively) and DNA isolated. The number of base lesions was determined as described above.

Finally, another group of derivatives are the aldehyde-/ketone-reactive probes. This group is based on the activation of a sulfonyl hydrazine group of carbon number 5. They are based on Lissamine and Texas Red structures and used to label aldehyde-/ketone-containing molecules (with sugars). The most common aldehyde-/ ketone-reactive probes are Lissamine rhodamine B hydrazine and Texas Red hydrazine. 

From this study it seems that oxygenated aromatic compounds are less reactive towards acidic CO formylation in comparison to alkylated ones and this effect can be explained by the relative acid-base properties of the substrate and carbon monoxide. More than stoichiometric amounts of acid are required for acceptable rates and yields. Various acids apart from the HF/BF3 system were investigated without success in an attempt to facilitate formylation of anisole. Deuterium labelling experiments as a tool to probe the possible reaction mechanism provided interesting but inconclusive results in support of the intra-complex mechanism. Secondary product formation was shown to result mainly from acid catalysed reactions between the substrate and the aldehyde product. Preliminary economic evaluations indicated boron trifluoride to be the major cost driver in these reactions, necessitating its quantitative recovery. A successful FIF/BF3 retrieval process is described by Fujiyama et and involves the decomposition of the p-... 

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