TMAO

Fig.l. Gene maps of recombinant plasmids pTOH, pTTOH, pEOH, and pETOH. Abbreviations Pnc, trc promoter Px7, T7 and lac hybrid promoter Tat, twin-arginme TorA signal sequence of TMAO reductase OPH, organophosphoms hydrolase gene Hisg, hexa-histidine affinity tag. 

The conditions under which these function and their regulation depend on the organism. For example, in Escherichia coli, oxygen represses the synthesis of the other reductases, and under anaerobic conditions the reductases for fumarate, DMSO, and TMAO are repressed by nitrate. This does not apply to Wolinella succinogenes in which sulfur represses the synthesis of the more positive electron acceptors nitrate and fumarate (Lorenzen et al. 1993). The DMSO reductase from Escherichia coli (Weiner et al. 1988) has a broad substrate versatility, and is able to reduce a range of sulfoxides and A-oxides. Anaerobic sulfate reduction is not discussed here in detail. 

Metabolism of trimethylamine oxide in fish muscle involves an enzyme-catalyzed oxidation-reduction reaction. The enzyme responsible for the conversion of trimethylamine oxide to trimethylamine is known as trimethylamine-W-oxide reductase. This enzyme acts on nicotinamide adenine dinucleotide (NADH) and TMAO to produce NAD+, trimethylamine and water (Fig. 13.13.1). TMAO acts as the oxidizing agent and is reduced, while NADH undergoes oxidation as the reducing agent. 

An alternate enzyme, trimethylamine-oxide aldolase, converts TMAO to dime-hylamine and formaldehyde, as written in Fig. 13.13.2. 

Main metabolites in urine after 7 months of exposure to 100 mg As/L drinking water were DMA and trimethylarsin oxide (TMAO) with minute amounts of tetramethylarsonium (TMA)... 

Trimethylarsin oxide (TMAO) 100 mg As/L in drinking water for 7 months was excreted in urine mostly unchanged with some TMA 24... 

An efficient and selective Cu-assisted ortho-hydroxylation procedure for the conversion of benzoate to salicylate has been described, involving trimethylamine N-oxide (TMAO) as the oxidant [191,192]. The reaction was assumed to proceed via oxidation of a Cu carboxylate complex by TMAO to produce the active species (postulated to be a Cu hydroxo complex, but with only circumstantial evidence), followed by oxygen transfer to the benzoate group (Scheme 14). Using a set of different amide derivatives of benzoic acid, Comba and co-workers gained additional mechanistic hints in support of a reactive Cu-oxo or Cu-hydroxo intermediate that is stabilized by a five-membered chelate [193]. A pre-equilibrium involving Cu the ligand, and TMAO was proposed, but details of the reaction are far from clear. 

Scheme 14 Cu-assisted ortho-hydroxylation of benzoate to salicylate with TMAO...

Another chemical method for measuring freshness, that is more rapid, continuous, and less destructive than other methods is the detection of volatile trimethylamine (TMA), dimethylamine (DMA), monomethylamine (MMA), and ammonia 14,15). Trimethylamine oxide (TMAO) is a decomposition product of proteins as well as present in excretions of fish 16). Spoilage bacteria can reduce TMAO to TMA plus small amounts of DMA, MMA, and ammonia. Tissue TMA levels have be correlated with the pungent odor associated with spoiled seafood as well as total bacterial counts 14). Researchers incorporated a test strip... 

Arsenic compounds with one to four methyl groups attached to the arsenic atom are common constituents of marine samples. The relevant species are monomethylarsonic acid (MMA) (3), dimethylarsinic acid (DMA) (4), trimethylarsine oxide (TMAO) (5), and tetramethylarson-ium ion (TeMA) (6). Of these, MMA and DMA are readily separated... 

The compounds MMA, DMA, and TMAO are reduced in acidic aqueous media by borohydride solutions to methylarsine (MeAsH2, bp 2°C), dimethylarsine (Me2AsH, bp 35°C), and trimethylarsine (Me3As, bp 55°C), respectively. These products are useful derivatives for speciation analysis of arsenic because they are readily separated from complex sample matrices and may be further separated from each other by distillation (41) or by gas chromatography (42) prior to their determination by element-specific detectors. Consequently, arsine generation techniques are the most commonly used methods for determining MMA, DMA, and TMAO in marine samples. 

The arsenic constituents of marine algae have been the subject of several detailed studies over the past 15 years. It is of interest that arsenobetaine, arsenocholine, TMAO, and TeMA are yet to be detected in marine algae. 

TMAO has been detected in fish (37, 112, 113), molluscs (37, 108), and crustaceans (37). It may occur from microbial decomposition of... 

According to Challenger (127), arsenate is transformed to trimethyl-arsine by the mold Scopulariopsis brevicaulis, by sequential reduction and oxidative methylation of the arsenic species (Fig. 7). The proposed intermediates in the pathway were MMA, DMA, and TMAO. Although Challenger could not detect these compounds, when they were added to a culture of S. brevicaulis trimethylarsine was formed. Challenger (129) considered that the likely source of methyl groups was S-adeno-sylmethionine (AdoMet), which had previously been identified as an... 

S. brevicaulis (134). By the use of sensitive analytical methodology involving arsine generation, the arsenic intermediates proposed in Fig. 7 were identified in the growth medium. A significant result was the detection of TMAO, rather than trimethylarsine, as the major methylation product. The low concentrations of arsenic employed in these experiments resulted in TMAO being present at less than toxic concentrations, and further detoxification by reducing TMAO to the arsine was considered unnecessary. Whether or not TeMA was produced in these experiments is not known its presence was not reported, but it would not have been detected by the analytical technique used. 

Hanaoka and co-workers (141) have reported several experiments describing the microbial degradation of arsenobetaine. Unspecified microorganisms derived from sediments, algae, mollusc intestine, or suspended sediments were incubated with arsenobetaine under various conditions. Arsenobetaine was degraded to TMAO and DMA in all cases and, for sediments and suspended sediments, further degradation... 

Uptake experiments have also been conducted with other forms of arsenic. M. edulis exposed separately to a number of organoarsenic compounds in seawater were found to be selective in their arsenic uptake (85). They did not accumulate arsenic when it was present as MMA, DMA, TMAO, or DMAE. When exposed to the three quaternary arsonium compounds (arsenobetaine, arsenocholine, and TeMA), however, the mussels accumulated substantial quantities of arsenic, with arsenobetaine being the most efficiently accumulated (Table VII). The chemical form of the accumulated arsenic was also examined. Mussels... 

Beside BAs, low-molecular-weight alkylamines, commonly used as indicators of food quality, can also be present in fish muscle. Tri- and dimethylamine (TMA and DMA) are produced by bacterial reduction of the osmoregulatory substance trimethylamine oxide (TMAO) in fresh marine fish and by enzymatic reduction in frozen storage of gadoid fish (cod, cusk, hake, pollack), with concurrent formation of formaldehyde. 

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