Growth and Arginine Metabolism of the Wine Lactic Acid Bacteria Lactobacillus buchneri and Oenococcus oeni at Different pH Values and Arginine Concentrations (original) (raw)
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Journal of Applied Microbiology, 2000
I L O N E . 2000. The excretion of citrulline, a precursor of carcinogenic ethyl carbamate, formed from arginine degradation by malolactic bacteria in wine is of toxicological concern. The arginine metabolism of resting cells of Lactobacillus buchneri CUC-3 and Oenococcus oeni Lo111 was examined. The citrulline excretion rate was found to be linearly correlated to the arginine degradation rate. It was possible to calculate an arginine to citrulline conversion ratio which could be used to predict the amount of citrulline expected after the degradation of a known quantity of arginine. The conversion ratios determined in this study were similar to data calculated from other authors for fermentations in wine and ranged between 4Á0% and 7Á7%. Ribose, fructose and glucose inhibited the degradation of arginine in Lact. buchneri CUC-3, and inhibition of arginine degradation by glucose correlated with higher arginine to citrulline conversion ratios. The work presents new results of arginine metabolism in malolactic bacteria and gives starting points for investigations in wine.
Influence of wine-like conditions on arginine utilization by lactic acid bacteria
International Microbiology, 2011
Wine can contain trace amounts of ethyl carbamate (EC), a carcinogen formed when ethanol reacts with carbamyl compounds such as citrulline. EC is produced from arginine by lactic acid bacteria (LAB), e.g., Lactobacillus and Pediococcus. Although the amounts of EC in wine are usually negligible, over the last few years there has been a slight but steady increase, as climate change has increased temperatures and alcohol levels have become proportionately higher, both of which favor EC formation. In this study, resting cells of LAB were used to evaluate the effects of ethanol, glucose, malic acid, and low pH on the ability of non-oenococcal strains of these bacteria to degrade arginine and excrete citrulline. Malic acid was found to clearly inhibit arginine consumption in all strains. The relation between citrulline produced and arginine consumed was clearly higher in the presence of ethanol (10-12 %) and at low pH (3.0), which is consistent with both the decreased amount of ornithine produced from arginine and the reduction in arginine degradation. In L. brevis and L. buchneri strains isolated from wine and beer, respectively, the synthesis of citrulline from arginine was highest.
Arginine, citrulline and ornithine metabolism by lactic acid bacteria from wine
International Journal of Food Microbiology, 1999
The catabolism of arginine, an amino acid found in grape juice and wine, citrulline and ornithine was investigated in four lactic acid bacteria. Only Lactobacillus hilgardii X B catabolized arginine and excreted citrulline into the medium. The 1 recovery of arginine as ornithine was lower than the expected theoretical value. The arginase-urease pathway was not detected indicating that the amino acid degradation was carried out only by the arginine dihydrolase pathway. Oenococcus oeni m, a strain not able to utilize arginine, degraded citrulline that was completely recovered as ornithine, ammonia and CO . Lactobacillus hilgardii X B catabolized citrulline but it was only 44% recovered as ornithine. The citrulline utilization 2 1 by Oenococcus oeni m may be important for two reasons: it can gain extra energy for growth from citrulline metabolism, and the amino-acid diminution could avoid the possibility of ethyl carbamate formation from the citrulline naturally present in wine.
Fems Microbiology Letters, 2000
Major commercially available strains for induction of malolactic fermentation in wine were examined for arginine metabolism in a resting cell system at wine pH with the aim of evaluating their ability to excrete and utilize citrulline, a precursor of carcinogenic ethyl carbamate (urethane). All strains tested excreted citrulline from arginine degradation. Citrulline was stored intracellularly during growth in arginine rich medium and was released upon lysis of the cells. All strains were found to degrade citrulline as a sole amino acid and some of them were able to reutilize previously excreted citrulline.
Research in Microbiology, 2005
The aim of this work was to study the effects of ethanol on cell growth and arginine and citrulline metabolism in two heterofermentative lactic acid bacteria from wine, and to determine their possible association with the formation of ethyl carbamate (EC), a carcinogenic compound. Lactobacillus hilgardii X 1 B is able to utilize arginine and citrulline, while Oenococcus oeni m can only use citrulline, a precursor of EC. Growth of both microorganisms was partially inhibited by 10 and 15% (v/v) ethanol. Specific arginine consumption by L. hilgardii increased when the pH value diminished from 6.5 to 3.8, but was not affected by an increasing ethanol concentration. However, the ethanol concentration affected the specific citrulline consumption of both microorganisms. Arginine metabolism by L. hilgardii X 1 B increased the amount of citrulline, thus allowing production of EC in the medium. Citrulline utilization by both microorganisms, at all pH values studied, indirectly inhibited the formation of EC; indeed, one of the precursors had practically disappeared after 48 h of incubation. Due to its ability to form precursors, L. hilgardii X 1 B has the potential to contribute to EC formation, whereas citrulline utilization by O. oeni m in the presence of ethanol may contribute to diminishing the formation of EC. Rapid degradation of citrulline in the presence of ethanol by O. oeni m is important from a toxicological point of view, because it is important to keep the EC levels as low as possible.
Metabolism of arginine and its positive effect on growth and revival of Oenococcus oeni
Journal of Applied Microbiology, 2000
Oenococcus oeni is the main lactic acid bacteria species which induces malolactic fermentation during wine-making. It is able to break down arginine via the arginine deiminase pathway, a potential source of energy already considered for many bacteria. The production of ATP by starved cells from arginine was quanti®ed with a bioluminescence assay, and ef®cient coupling of amino acid catabolism and cell growth was monitored. Therefore, molecular growth yield was determined after glucose exhaustion. With colony plate counting and a direct epi¯uorescence technique, it was shown that addition of arginine to viable but non-culturable cells obtained after nutrient starvation restored their ability to grow during its degradation. Therefore, arginine produced more than maintenance energy. It is concluded that strains which are able to metabolize arginine might take advantage of this additional energy source for growth.
Arginine metabolism by wine Lactobacilli isolated from wine
Food Microbiology, 2002
Lactobacillus hilgardii is a very common heterofermentative bacterium found in wine, associated mainly with several kinds of negative alterations. It is also known as a spoilage organism in soft drinks and other fermented beverages. It is able to break down arginine, one of the most abundant amino acids in wine, through the arginine deiminase (ADI) pathway.The ¢rst step of this metabolism may lead to the excretion of citrulline. This feature has an important enological implication since citrulline can react spontaneously with ethanol to form ethyl carbamate. Carcinogenic e¡ects of this compound have been observed when administrated at high concentrations to laboratory animals. To complete the understanding of this catabolic pathway in L. hilgardii, arginine and citrulline utilization were investigated under di¡erent conditions. Moreover, ATP production from these amino acids was also monitored. From these data, arginine degradation via the ADI pathway can be considered to be a mechanism of energy production and pH regulation. However, if arginine degradation is bene¢cial for the bacteria, improving its growing ability and its adaptability, this increases the risk of degradation of the organoleptic and hygienic properties of wine.
Preliminary characterization of wine lactobacilli able to degrade arginine
World Journal of Microbiology & Biotechnology, 2002
Lactobacillus strains able to degrade arginine were isolated and characterized from a typical red wine. All the strains were gram-positive, catalase-negative and produced both D -and L -lactate from glucose. Strains L2, L3, L4, and L6 were able to produce CO 2 from glucose; however, production of CO 2 from glucose was not observed in strains L1 and L5, suggesting that they belong to the homofermentative wine lactic acid bacteria (LAB) group. All of the lactobacilli were tested for their ability to ferment 49 carbohydrates. The sugar fermentation profile of strain L1 was unique, suggesting that this strain belonged to Lactococcus lactis ssp. cremoris, a non-typical wine LAB. Furthermore, a preliminary typing was performed by using a random amplified polymorphic DNA analysis (RAPD-PCR analysis).
Relationships between arginine degradation, pH and survival in Lactobacillus sakei
FEMS Microbiology Letters, 1999
Lactobacillus sakei is one of the most important lactic acid bacteria of meat and fermented meat products. It is able to degrade arginine with ammonia and ATP production by the arginine deiminase pathway (ADI). This pathway is composed of three enzymes: arginine deiminase, ornithine transcarbamoylase and carbamate kinase, and an arginine transport system. The transcription of the ADI pathway is induced by arginine and subjected to catabolite repression. In order to understand the physiological role of the degradation of this amino acid we investigated the growth of bacteria under various conditions. We show that arginine degradation is responsible for an enhanced viability during the stationary phase when cells are grown under anaerobiosis. Arginine is necessary for the induction of the ADI pathway but in association with another environmental signal. Using a mutant of the L-lactate dehydrogenase unable to lower the pH we could clearly demonstrate that (i) low pH is not responsible for cell death during the stationary phase, so survival is due to another factor than elevated pH, (ii) neither low pH nor oxygen limitation is responsible for the induction of the ADI pathway together with arginine since the ldhL mutant is able to degrade arginine under aerobiosis.
International Journal of Food Microbiology, 2009
Sourdough lactic acid bacteria (LAB) need to be adapted to a highly acidic and, therefore, challenging environment. Different mechanisms are employed to enhance competitiveness, among which conversion of arginine into ornithine through the arginine deiminase (ADI) pathway is an important one. A combined molecular and kinetic approach of the ADI pathway in Lactobacillus fermentum IMDO 130101, a highly competitive sourdough LAB strain, identified mechanisms with advantageous technological effects and quantified the impact of these effects. First, molecular analysis of the arcBCAD operon of 4.8 kb revealed the genes encoding the enzymes ornithine transcarbamoylase, carbamate kinase, arginine deiminase, and an arginine/ornithine (A/O) antiporter, respectively, with an additional A/O antiporter 702.5 kb downstream of the ADI operon. The latter could play a role in citrulline transport. Second, pH-controlled batch fermentations were carried out, generating data for the development of a mathematical model to describe the temporal evolution of the three amino acids involved in the ADI pathway (arginine, citrulline, and ornithine) as a result of the activity of these enzymes and transporter(s). Free arginine in the medium was converted completely into a mixture of citrulline and ornithine under all conditions tested. However, the ratio between these end-products and the pattern of their formation showed variation as a function of environmental pH. Under optimal pH conditions for growth, citrulline release and some further conversion into ornithine was observed. When growing under sub-optimal pH conditions, ornithine was the main product of the ADI pathway. These kinetic data suggest a role in adaptation of L. fermentum IMDO 130101 to growth under suboptimal conditions.