Furfural inhibits growth by limiting sulfur assimilation in ethanologenic Escherichia coli strain LY180.

A wide variety of commercial products can be potentially made from monomeric sugars produced by the dilute acid hydrolysis of lignocellulosic biomass. However, this process is accompanied by side products such as furfural that hinder microbial growth and fermentation. To investigate the mechanism of furfural inhibition, mRNA microarrays of an ethanologenic strain of Escherichia coli (LY180) were compared immediately prior to and 15 min after a moderate furfural challenge. Expression of genes and regulators associated with the biosynthesis of cysteine and methionine was increased by furfural, consistent with a limitation of these critical metabolites. This was in contrast to a general stringent response and decreased expression of many other biosynthetic genes. Of the 20 amino acids individually tested as supplements (100 microM each), cysteine and methionine were the most effective in increasing furfural tolerance with serine (precursor of cysteine), histidine, and arginine of lesser benefit. Supplementation with other reduced sulfur sources such as d-cysteine and thiosulfate also increased furfural tolerance. In contrast, supplementation with taurine, a sulfur source that requires 3 molecules of NADPH for sulfur assimilation, was of no benefit. Furfural tolerance was also increased by inserting a plasmid encoding pntAB, a cytoplasmic NADH/NADPH transhydrogenase. Based on these results, a model is proposed for the inhibition of growth in which the reduction of furfural by YqhD, an enzyme with a low K(m) for NADPH, depletes NADPH sufficiently to limit the assimilation of sulfur into amino acids (cysteine and methionine) by CysIJ (sulfite reductase).

Inhibition of sulfur incorporation to transfer RNA by ultraviolet-A radiation in Escherichia coli.

tRNA sulfurtransferase activity was assayed in Escherichia coli cell extracts obtained from bacterial suspensions exposed to a sub-lethal dose of ultraviolet-A radiation (fluence 148 kJ m(-2)) imparted at a low fluence rate (41 W m(-2)). We found that the irradiation reduced the enzymatic activity to one fourth of the control value, indicating that ultraviolet-A exposure inhibits the synthesis of 4-thiouridine, the most abundant thionucleoside in E. coli tRNA. Changes in the tRNA content of 4-thiouridine and its derived photoproduct 5-(4'-pyrimidin 2'-one) cytosine were studied in bacteria growing under ultraviolet-A irradiation. In these conditions the accumulation of photoproduct was limited, and the kinetics of this process was non-coincident with disappearance of 4-thiouridine. The results, which are compatible with the fact that ultraviolet-A induces an inhibition of the 4-thiouridine synthesis, suggest that the effect of radiation on tRNA modification is relevant to tRNA photo-inactivation in growing bacteria.

The effect of zinc-containing chewing gum on volatile sulfur-containing compounds in the oral cavity.

Volatile sulfur-containing compounds (VSC) are known to constitute the major component of halitosis. Aqueous solutions of zinc salts have been shown to reduce the levels of VSC produced orally. The aim of the present study was to examine whether zinc could be made available in the oral cavity and inhibit VSC production when delivered by a chewing gum. VSC measurements were carried out on the 'morning breath' of 11 test subjects and re-examined after the use of test solutions containing 0.02% zinc chloride, 0.2% chlorhexidine, or water or the use of chewing gums containing 2 mg, 0.5 mg, or 0 mg zinc acetate. The results showed that similar amounts of zinc in mouthrinses or chewing gum had the same effect, with a reduction of the oral VSC of 45%. Chewing gum thus seems to be a viable alternative for delivering zinc to reduce VSC levels in the oral cavity.

Effect of high-level copper feeding on the sulfur amino acid need of chicks fed corn-soybean meal and purified crystalline amino acid diets.

Supplementation of a corn-soybean meal diet with .05% methionine maximized gain and feed efficiency of three-week-old chicks fed copper at either 0 or 250 mg/kg. With copper present at 500 mg/kg, the supplemental methionine requirement was in excess of .10%. Liver copper stores of chicks fed copper at 250 mg/kg declined linearly as supplemental methionine level increased, but methionine level had no effect on liver copper when chicks were fed 500 mg/kg copper. Liver copper accumulation did not occur in chicks fed 250 mg/kg copper when supplemental sulfur was provided as a combination of .05% methionine and 1800 mg/kg sulfide. Irrespective of methionine level, sulfide addition reduced liver copper by one-half in birds fed 500 mg/kg copper. Chicks fed the purified diet were appreciably more susceptible to copper toxicity than those fed the corn-soybean meal diet. Chicks fed the purified diet without supplemental copper required .52% sulfur amino acids (SAA). Chicks fed copper at 250 and 500 mg/kg required .65 and .70 SAA, respectively, although gain and feed efficiency were still depressed when compared with the performance of the control birds. Higher levels of methionine failed to restore growth rates to those achieved with birds fed no supplemental copper. Supplementing the copper-containing diets with up to 1.6% cystine improved performance some but failed to completely overcome the growth depressing effect of copper at 500 mg/kg. Results of additional studies indicated that the purified diet containing .70% SAA and 500 mg/kg copper was nutritionally adequate and that the marked toxicity of copper when fed in the purified diet was due to the chemical nature of the diet rather than to any specific copper-induced nutritional deficiencies.