Bacteria do not incorporate ß-N-methylamino-L-alanine into their proteins.

ß-N-methylamino-l-alanine (BMAA), is commonly found in both a free and proteinassociated form in various organisms exposed to the toxin. The long latency of development of neurodegeneration attributed to BMAA, is hypothesized to be the result of excitotoxicity following slow release of the toxin from protein reservoirs. It was recently suggested that these BMAA-protein associations may reflect misincorporation of BMAA in place of serine, as occurs, for example, when canavanine misincorporates in place of arginine. We therefore compared BMAA and canavanine toxicty in various bacterial species, and misincorporation of these amino acids into proteins in a bacterial protein expression system. None of the bacterial species showed any physiological stress responses to BMAA in contrast to the growth reduction observed when cultures were incubated in media containing canavanine. LC-MS analysis confirmed uptake of BMAA from growth media. However, after immobilized metal affinity chromatography and SDS-PAGE purification of proteins produced in an E scherichia coli expression system, no BMAA was detected by either LC-MS or LC-MS/MS analysis using two derivatization methods, or by orbitrap MS of trypsin digests of the protein. We therefore conclude that BMAA is not misincorporated into proteins in bacteria and that the observed BMAA-protein association in bacteria is superficial.

The localization of exogenous microcystin LR taken up by a non-microcystin producing cyanobacterium.

The effect of exogenous microcystin on non-microcystin producing cyanobacteria has not yet been extensively studied. Existing evidence for internalization of microcystin by cyanobacteria is based only on the presence of internalized radioisotopic label. Where a function or physiological role for microcystin has been proposed based on the molecule acting as a signalling molecule, the hypothetical function has not been demonstrated at the site of action in receiving cells. We therefore exposed Synechocystis PCC6803 to microcystin LR and showed that the microcystin-LR was both taken up by Synechocystis PCC6803 and localised in the thylakoid membranes, where it caused a decrease in photosystem II activity as has been shown for endogenous microcystin, without any negative effects on the cell's survival.

A GROWTH ADVANTAGE FOR MICROCYSTIN PRODUCTION BY MICROCYSTIS PCC7806 UNDER HIGH LIGHT(1).

Batch cultures of both Microcystis PCC7806 and a mcyA(-) knockout mutant (MT) of PCC7806 were cultured at three different light intensities and five media treatments, so as to vary cellular N:C ratios and concentrations and sampled daily over 5 d for analysis of microcystin concentration, cell numbers, and residual nitrate in the growth medium. A competitive survival advantage was noted at a high-light level (37 µmol photons · m(-2 ) · s(-1) ), where the toxic strain survived while the nontoxic strain became chlorotic. A strong correlation (r(2) = 0.91, P < 0.001, N = 22) between microcystin concentration and growth rate was observed at high-light conditions. No advantage was observed at optimal or low-light conditions, and media composition had no significant effect on the relationship between toxicity and survival at high-light conditions. These data suggest a possible role for microcystin in protection against photooxidation.