scholarly journals In Lactobacillus plantarum, Carbamoyl Phosphate Is Synthesized by Two Carbamoyl-Phosphate Synthetases (CPS): Carbon Dioxide Differentiates the Arginine-Repressed from the Pyrimidine-Regulated CPS

2000 ◽  
Vol 182 (12) ◽  
pp. 3416-3422 ◽  
Author(s):  
Hervé Nicoloff ◽  
Jean-Claude Hubert ◽  
Françoise Bringel
Keyword(s):  
Carbon Dioxide ◽  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Lactobacillus Plantarum ◽  
Bicarbonate Level ◽  
Carbamoyl Phosphate Synthetase ◽  
Wild Type ◽  
Arginine Biosynthesis ◽  
Carbamoyl Phosphate ◽  
Double Deletion

ABSTRACT Carbamoyl phosphate (CP) is an intermediate in pyrimidine and arginine biosynthesis. Carbamoyl-phosphate synthetase (CPS) contains a small amidotransferase subunit (GLN) that hydrolyzes glutamine and transfers ammonia to the large synthetase subunit (SYN), where CP biosynthesis occurs in the presence of ATP and CO2.Lactobacillus plantarum, a lactic acid bacterium, harbors a pyrimidine-inhibited CPS (CPS-P; Elagöz et al., Gene 182:37–43, 1996) and an arginine-repressed CPS (CPS-A). Sequencing has shown that CPS-A is encoded by carA (GLN) and carB (SYN). Transcriptional studies have demonstrated that carB is transcribed both monocistronically and in the carABarginine-repressed operon. CP biosynthesis in L. plantarumwas studied with three mutants (ΔCPS-P, ΔCPS-A, and double deletion). In the absence of both CPSs, auxotrophy for pyrimidines and arginine was observed. CPS-P produced enough CP for both pathways. In CO2-enriched air but not in ordinary air, CPS-A provided CP only for arginine biosynthesis. Therefore, the uracil sensitivity observed in prototrophic wild-type L. plantarum without CO2 enrichment may be due to the low affinity of CPS-A for its substrate CO2 or to regulation of the CP pool by the cellular CO2/bicarbonate level.

scholarly journals Repression of the pyr Operon in Lactobacillus plantarum Prevents Its Ability To Grow at Low Carbon Dioxide Levels

2005 ◽  
Vol 187 (6) ◽  
pp. 2093-2104 ◽  
Author(s):  
Hervé Nicoloff ◽  
Aram Elagöz ◽  
Florence Arsène-Ploetze ◽  
Benoît Kammerer ◽  
Jan Martinussen ◽  
...  
Keyword(s):  
Carbon Dioxide ◽  
Lactobacillus Plantarum ◽  
De Novo ◽  
Site Directed Mutagenesis ◽  
Low Carbon ◽  
Wild Type ◽  
Carbamoyl Phosphate ◽  
Phosphoribosyl Pyrophosphate ◽  
Pyrimidine Nucleotides ◽  
Transcription Attenuation

ABSTRACT Carbamoyl phosphate is a precursor for both arginine and pyrimidine biosynthesis. In Lactobacillus plantarum, carbamoyl phosphate is synthesized from glutamine, ATP, and carbon dioxide by two sets of identified genes encoding carbamoyl phosphate synthase (CPS). The expression of the carAB operon (encoding CPS-A) responds to arginine availability, whereas pyrAaAb (encoding CPS-P) is part of the pyrR1BCAaAbDFE operon coding for the de novo pyrimidine pathway repressed by exogenous uracil. The pyr operon is regulated by transcription attenuation mediated by a trans-acting repressor that binds to the pyr mRNA attenuation site in response to intracellular UMP/phosphoribosyl pyrophosphate pools. Intracellular pyrimidine triphosphate nucleoside pools were lower in mutant FB335 (carAB deletion) harboring only CPS-P than in the wild-type strain harboring both CPS-A and CPS-P. Thus, CPS-P activity is the limiting step in pyrimidine synthesis. FB335 is unable to grow in the presence of uracil due to a lack of sufficient carbamoyl phosphate required for arginine biosynthesis. Forty independent spontaneous FB335-derived mutants that have lost regulation of the pyr operon were readily obtained by their ability to grow in the presence of uracil and absence of arginine; 26 harbored mutations in the pyrR1-pyrB loci. One was a prototroph with a deletion of both pyrR1 and the transcription attenuation site that resulted in large amounts of excreted pyrimidine nucleotides and increased intracellular UTP and CTP pools compared to wild-type levels. Low pyrimidine-independent expression of the pyr operon was obtained by antiterminator site-directed mutagenesis. The resulting AE1023 strain had reduced UTP and CTP pools and had the phenotype of a high-CO2-requiring auxotroph, since it was able to synthesize sufficient arginine and pyrimidines only in CO2-enriched air. Therefore, growth inhibition without CO2 enrichment may be due to low carbamoyl phosphate pools from lack of CPS activity.

scholarly journals Two Arginine Repressors Regulate Arginine Biosynthesis in Lactobacillus plantarum

2004 ◽  
Vol 186 (18) ◽  
pp. 6059-6069 ◽  
Author(s):  
Hervé Nicoloff ◽  
Florence Arsène-Ploetze ◽  
Cédric Malandain ◽  
Michiel Kleerebezem ◽  
Françoise Bringel
Keyword(s):  
Lactobacillus Plantarum ◽  
Wild Type ◽  
Arginine Biosynthesis ◽  
Carbamoyl Phosphate ◽  
Gram Positive ◽  
E Coli ◽  
Gram Positive Bacteria ◽  
Arginine Repressor ◽  
In The Wild ◽  
Spontaneous Mutants

ABSTRACT The repression of the carAB operon encoding carbamoyl phosphate synthase leads to Lactobacillus plantarum FB331 growth inhibition in the presence of arginine. This phenotype was used in a positive screening to select spontaneous mutants deregulated in the arginine biosynthesis pathway. Fourteen mutants were genetically characterized for constitutive arginine production. Mutations were located either in one of the arginine repressor genes (argR1 or argR2) present in L. plantarum or in a putative ARG operator in the intergenic region of the bipolar carAB-argCJBDF operons involved in arginine biosynthesis. Although the presence of two ArgR regulators is commonly found in gram-positive bacteria, only single arginine repressors have so far been well studied in Escherichia coli or Bacillus subtilis. In L. plantarum, arginine repression was abolished when ArgR1 or ArgR2 was mutated in the DNA binding domain, or in the oligomerization domain or when an A123D mutation occurred in ArgR1. A123, equivalent to the conserved residue A124 in E. coli ArgR involved in arginine binding, was different in the wild-type ArgR2. Thus, corepressor binding sites may be different in ArgR1 and ArgR2, which have only 35% identical residues. Other mutants harbored wild-type argR genes, and 20 mutants have lost their ability to grow in normal air without carbon dioxide enrichment; this revealed a link between arginine biosynthesis and a still-unknown CO2-dependent metabolic pathway. In many gram-positive bacteria, the expression and interaction of different ArgR-like proteins may imply a complex regulatory network in response to environmental stimuli.

scholarly journals Extent of Genetic Lesions of the Arginine and Pyrimidine Biosynthetic Pathways in Lactobacillus plantarum, L. paraplantarum, L. pentosus, and L. casei: Prevalence of CO2-Dependent Auxotrophs and Characterization of Deficient arg Genes in L. plantarum

2003 ◽  
Vol 69 (5) ◽  
pp. 2674-2683 ◽  
Author(s):  
Françoise Bringel ◽  
Jean-Claude Hubert
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Single Point ◽  
Point Mutations ◽  
Pyrimidine Biosynthesis ◽  
Arginine Biosynthesis ◽  
Carbamoyl Phosphate ◽  
Random Mutation ◽  
Rich Media

ABSTRACT Lactic acid bacteria require rich media since, due to mutations in their biosynthetic genes, they are unable to synthesize numerous amino acids and nucleobases. Arginine biosynthesis and pyrimidine biosynthesis have a common intermediate, carbamoyl phosphate (CP), whose synthesis requires CO2. We investigated the extent of genetic lesions in both the arginine biosynthesis and pyrimidine biosynthesis pathways in a collection of lactobacilli, including 150 strains of Lactobacillus plantarum, 32 strains of L. pentosus, 15 strains of L. paraplantarum, and 10 strains of L. casei. The distribution of prototroph and auxotroph phenotypes varied between species. All L. casei strains, no L. paraplantarum strains, two L. pentosus strains, and seven L. plantarum strains required arginine for growth. Arginine auxotrophs were more frequently found in L. plantarum isolated from milk products than in L. plantarum isolated from fermented plant products or humans; association with dairy products might favor arginine auxotrophy. In L. plantarum the argCJBDF genes were functional in most strains, and when they were inactive, only one gene was mutated in more than one-half of the arginine auxotrophs. Random mutation may have generated these auxotrophs since different arg genes were inactivated (there were single point mutations in three auxotrophs and nonrevertible genetic lesions in four auxotrophs). These data support the hypothesis that lactic acid bacteria evolve by progressively loosing unnecessary genes upon adaptation to specific habitats, with genome evolution towards cumulative DNA degeneration. Although auxotrophy for only uracil was found in one L. pentosus strain, a high CO2 requirement (HCR) for arginine and pyrimidine was common; it was found in 74 of 207 Lactobacillus strains tested. These HCR auxotrophs may have had their CP cellular pool-related genes altered or deregulated.

scholarly journals Draft Genome Sequence of Lactobacillus plantarum SF2A35B

2016 ◽  
Vol 4 (1) ◽  
Author(s):  
Peter A. Bron ◽  
I-Chiao Lee ◽  
Lennart Backus ◽  
Sacha A. F. T. van Hijum ◽  
Michiel Wels ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Lactobacillus Plantarum ◽  
Genome Sequence ◽  
Draft Genome ◽  
Draft Genome Sequence

The lactic acid bacterium Lactobacillus plantarum is intensively studied as a model probiotic species. Here, we present the draft genome sequence of the exopolysaccharide-producing strain SF2A35B.

Preserving Herbs with Lactobacillus plantarum-52 Lactic-acid Bacterium and Their use in Forage For Cows

2015 ◽  
Vol 12 (1) ◽  
pp. 231-235
Author(s):  
Zh. Ibraimova ◽  
A. Rustenov ◽  
R. Abildaeva ◽  
A. Dauylbay ◽  
A. Saparbekova
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Lactobacillus Plantarum

scholarly journals The Pyrimidine Operon pyrRPB-carA fromLactococcus lactis

2001 ◽  
Vol 183 (9) ◽  
pp. 2785-2794 ◽  
Author(s):  
Jan Martinussen ◽  
Jette Schallert ◽  
Birgit Andersen ◽  
Karin Hammer
Keyword(s):  
Mutational Analysis ◽  
Regulatory Gene ◽  
Regulatory Protein ◽  
Small Subunit ◽  
Transcriptional Level ◽  
Carbamoyl Phosphate Synthetase ◽  
Arginine Biosynthesis ◽  
Carbamoyl Phosphate ◽  
Biosynthetic Genes ◽  
Membrane Bound

ABSTRACT The four genes pyrR, pyrP, pyrB, and carAwere found to constitute an operon in Lactococcus lactissubsp. lactis MG1363. The functions of the different genes were established by mutational analysis. The first gene in the operon is the pyrimidine regulatory gene, pyrR, which is responsible for the regulation of the expression of the pyrimidine biosynthetic genes leading to UMP formation. The second gene encodes a membrane-bound high-affinity uracil permease, required for utilization of exogenous uracil. The last two genes in the operon, pyrBand carA, encode pyrimidine biosynthetic enzymes; aspartate transcarbamoylase (pyrB) is the second enzyme in the pathway, whereas carbamoyl-phosphate synthetase subunit A (carA) is the small subunit of a heterodimeric enzyme, catalyzing the formation of carbamoyl phosphate. The carAgene product is shown to be required for both pyrimidine and arginine biosynthesis. The expression of the pyrimidine biosynthetic genes including the pyrRPB-carA operon is subject to control at the transcriptional level, most probably by an attenuator mechanism in which PyrR acts as the regulatory protein.

Sign in / Sign up

Export Citation Format

Share Document