Ch as Lactobacillus sakei [13] and Lactobacillus delbrueckii [14]. Acetylation of the muramic

Ch as Lactobacillus sakei [13] and Lactobacillus delbrueckii [14]. Acetylation of the muramic and glucosamine residues of the peptidoglycan for instance, involves O-acetylation for which a supply of C2 compounds like acetyl-CoA is essential [15]. Heterofermentative lactic acid bacteria have the capacity for acetate production, and are therefore assumed to be independent of exogenous acetate addition. However, growth of a DLDHLactococcus lactis mutant was reported to be stimulated by acetateOxygen Effect on Lactobacillus Growth Requirementswhich it uses for the conversion to ethanol as a means to regenerate NAD+ in order to rescue its redox balance [16]. Another well-described growth requirement is CO2. L. johnsonii, is a so called capnophilic organism, i.e. it has a requirement for either gaseous CO2 or bicarbonate supplementation for growth, which is a characteristic that is also observed in many other lactic acid bacteria species [17?9]. The C-1 source has been proposed to be required for the synthesis of a common intermediate of the pyrimidine and arginine production pathways, carbamoyl-phosphate. In L. plantarum carbamoyl-phosphate can be synthesized from glutamine, ATP and bicarbonate involving two enzymes: pyrimidine-regulated CPS-P (encoded by carAB) and arginine regulated CPS-A (encoded by pyrAaAb) [20]. Two regulators of this tert-Butylhydroquinone pathway, PyrR1 and PyrR2 control 115103-85-0 expression of the pyr-operon in response to pyrimidine and inorganic carbon levels, respectively [21,22]. The genes of the pyr-operon 1662274 are conserved amongst many lactobacilli, including L, johnsonii NCC 533. Homologues of the argFGH genes for arginine biosynthesis are absent, rendering this species auxotrophic for arginine. The production and consumption of metabolites, like CO2 and acetate, are known to stabilize microbial communities. For example, in yoghurt fermentation, Streptococcus thermophilus and L. delbrueckii show close metabolic relations with the first species providing the second with CO2, acetate, folate, and formate. In exchange, the streptococcal species profits from the proteolytic activities of L. delbrueckii [23]. Analogously, it can be anticipated that specific nutritional requirements of microbes play an important role in the composition of the human microbiota. In view of both its industrial potential and its niche in the complex microbial environments where these lactobacilli are generally found, such as the gut, understanding the mechanisms that underlie these growth requirements are important. Growth requirements may be strongly dependent on the growth conditions. For L. johnsonii NCC 533 we observed major differences in growth and viability between aerobic and anaerobic conditions, including a significantly higher viability in the presence of molecular oxygen. This is surprising in view of the observation that L. johnsonii is known to produce hydrogen peroxide under aerobic conditions, a compound that is generally assumed to be toxic [24]. The study presented here indicates that the anaerobic dependency of L. johnsonii for carbon dioxide and acetate is related to its limited flexibility in pyruvate dissipation pathways, which can be overcome by pyruvate oxidase activity in the presence of oxygen, placing this enzyme in a pivotal position in the central metabolism of L. johnsonii.vacuuming, followed by replacement with the gas mixture of choice, and repeated 3 times at the start of the experiment as well as after every opening of the jar for sampling.Ch as Lactobacillus sakei [13] and Lactobacillus delbrueckii [14]. Acetylation of the muramic and glucosamine residues of the peptidoglycan for instance, involves O-acetylation for which a supply of C2 compounds like acetyl-CoA is essential [15]. Heterofermentative lactic acid bacteria have the capacity for acetate production, and are therefore assumed to be independent of exogenous acetate addition. However, growth of a DLDHLactococcus lactis mutant was reported to be stimulated by acetateOxygen Effect on Lactobacillus Growth Requirementswhich it uses for the conversion to ethanol as a means to regenerate NAD+ in order to rescue its redox balance [16]. Another well-described growth requirement is CO2. L. johnsonii, is a so called capnophilic organism, i.e. it has a requirement for either gaseous CO2 or bicarbonate supplementation for growth, which is a characteristic that is also observed in many other lactic acid bacteria species [17?9]. The C-1 source has been proposed to be required for the synthesis of a common intermediate of the pyrimidine and arginine production pathways, carbamoyl-phosphate. In L. plantarum carbamoyl-phosphate can be synthesized from glutamine, ATP and bicarbonate involving two enzymes: pyrimidine-regulated CPS-P (encoded by carAB) and arginine regulated CPS-A (encoded by pyrAaAb) [20]. Two regulators of this pathway, PyrR1 and PyrR2 control expression of the pyr-operon in response to pyrimidine and inorganic carbon levels, respectively [21,22]. The genes of the pyr-operon 1662274 are conserved amongst many lactobacilli, including L, johnsonii NCC 533. Homologues of the argFGH genes for arginine biosynthesis are absent, rendering this species auxotrophic for arginine. The production and consumption of metabolites, like CO2 and acetate, are known to stabilize microbial communities. For example, in yoghurt fermentation, Streptococcus thermophilus and L. delbrueckii show close metabolic relations with the first species providing the second with CO2, acetate, folate, and formate. In exchange, the streptococcal species profits from the proteolytic activities of L. delbrueckii [23]. Analogously, it can be anticipated that specific nutritional requirements of microbes play an important role in the composition of the human microbiota. In view of both its industrial potential and its niche in the complex microbial environments where these lactobacilli are generally found, such as the gut, understanding the mechanisms that underlie these growth requirements are important. Growth requirements may be strongly dependent on the growth conditions. For L. johnsonii NCC 533 we observed major differences in growth and viability between aerobic and anaerobic conditions, including a significantly higher viability in the presence of molecular oxygen. This is surprising in view of the observation that L. johnsonii is known to produce hydrogen peroxide under aerobic conditions, a compound that is generally assumed to be toxic [24]. The study presented here indicates that the anaerobic dependency of L. johnsonii for carbon dioxide and acetate is related to its limited flexibility in pyruvate dissipation pathways, which can be overcome by pyruvate oxidase activity in the presence of oxygen, placing this enzyme in a pivotal position in the central metabolism of L. johnsonii.vacuuming, followed by replacement with the gas mixture of choice, and repeated 3 times at the start of the experiment as well as after every opening of the jar for sampling.