Growth of Lactobacillus bulgaricus in Milk. 2. Characteristics of Purine Nucleotides, Pyrimidine Nucleotides, and Nucleic Acid Synthesis

Suzuki, Ichirou; Kato, Shintaro; Kitada, Tokuzou; Yano, Nobuhiro; Morichi, Toshiki

doi:10.3168/jds.s0022-0302(86)80490-8

Cited by 26 publications

(13 citation statements)

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“…An exception was pyrD, which was overexpressed in milk-grown cultures. It seems to reflect a direct influx of orotate present in milk (40). The tiled microarrays, however, identified three other genes that were overexpressed in the milk-grown cultures, cdc8/cmk, rtpR, and nrdD, that were predicted to encode thymidylate/cytidylate kinase; ribonucleotide-triphosphate reductase, class II (3); and ribonucleosidediphosphate reductase, class III (43), respectively, enzymes for the pyrimidine salvage pathway.…”

Section: Discussionmentioning

confidence: 99%

Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium

Smeianov

Wechter

Broadbent

et al. 2007

Appl Environ Microbiol

103

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Lactobacillus helveticus CNRZ32 is used by the dairy industry to modulate cheese flavor. The compilation of a draft genome sequence for this strain allowed us to identify and completely sequence 168 genes potentially important for the growth of this organism in milk or for cheese flavor development. The primary aim of this study was to investigate the expression of these genes during growth in milk and MRS medium by using microarrays. Oligonucleotide probes against each of the completely sequenced genes were compiled on maskless photolithography-based DNA microarrays. Additionally, the entire draft genome sequence was used to produce tiled microarrays in which noninterrupted sequence contigs were covered by consecutive 24-mer probes and associated mismatch probe sets. Total RNA isolated from cells grown in skim milk or in MRS to mid-log phase was used as a template to synthesize cDNA, followed by Cy3 labeling and hybridization. An analysis of data from annotated gene probes identified 42 genes that were upregulated during the growth of CNRZ32 in milk (P < 0.05), and 25 of these genes showed upregulation after applying Bonferroni's adjustment. The tiled microarrays identified numerous additional genes that were upregulated in milk versus MRS. Collectively, array data showed the growth of CNRZ32 in milk-induced genes encoding cell-envelope proteinases, oligopeptide transporters, and endopeptidases as well as enzymes for lactose and cysteine pathways, de novo synthesis, and/or salvage pathways for purines and pyrimidines and other functions. Genes for a hypothetical phosphoserine utilization pathway were also differentially expressed. Preliminary experiments indicate that cheesederived, phosphoserine-containing peptides increase growth rates of CNRZ32 in a chemically defined medium. These results suggest that phosphoserine is used as an energy source during the growth of L. helveticus CNRZ32.

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Section: Discussionmentioning

confidence: 99%

Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium

Smeianov

Wechter

Broadbent

et al. 2007

Appl Environ Microbiol

103

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“…For example, some LAB associated with particular fermented foods have developed auxotrophies for specific growth factors, including orotic acid present in milk for Lactobacillus delbrueckii subsp. bulgaricus (32), a small peptide present in freshly prepared yeast extracts for the sourdough bacterium Lactobacillus sanfrancisco (1), and D-mevalonic acid for rice wine spoilage lactobacilli (33). Some authors (21) consider LAB to be a highly specialized form of life in view of their complex nutritional requirements and restricted habitats.…”

mentioning

confidence: 99%

Extent of Genetic Lesions of the Arginine and Pyrimidine Biosynthetic Pathways in Lactobacillus plantarum , L. paraplantarum, L. pentosus , and L. casei : Prevalence of CO ₂ -Dependent Auxotrophs and Characterization of Deficient arg Genes in L. plantarum

Bringel

Hubert

2003

Appl Environ Microbiol

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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 CO 2 . 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 CO 2 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.Lactic acid bacteria (LAB) are gram-positive bacteria that have adapted to rich environments. As a result, in addition to sugars as energy and carbon sources these organisms require nucleobases, vitamins, cations, and amino acids (18). For example, some LAB associated with particular fermented foods have developed auxotrophies for specific growth factors, including orotic acid present in milk for Lactobacillus delbrueckii subsp. bulgaricus (32), a small peptide present in freshly prepared yeast extracts for the sourdough bacterium Lactobacillus sanfrancisco (1), and D-mevalonic acid for rice wine spoilage lactobacilli (33). Some authors (21) consider LAB to be a highly specialized form of life in view of their complex nutritional requirements and restricted habitats. The complex nutritional needs of LAB may be the result of two opposing evolutionary processes. A primitive LAB may have had restricted metabolism and gradually acquired new enzymatic activities. Alternatively, a chemoorganotrophic ancestor with many biochemical abilities may have evolved by progressively loosing unnecessary genes upon closer association with plants, animals, or hu...

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“…In other LAB, the occurrence of auxotrophies was correlated with specific growth factors present in a given habitat, including orotic acid present in milk for Lactobacillus delbrueckii subsp. bulgaricus [15] and D-mevalonic acid for rice wine spoilage lactobacilli [16]. In milk, the amounts of branched-chain amino acids are growth-limiting.…”

Section: Analysis Of the Nutritio-nal Needs With Respect To Strain Ormentioning

confidence: 99%

Article

Bringel¹,

Hubert²

2003

Lait

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-Lactic acid bacteria require rich media since, due to mutations in their biosynthetic genes, they are unable to synthesise numerous amino acids and nucleobases. The extent of genetic lesions was investigated in two biosynthetic pathways for 150 Lactobacillus plantarum isolates from various origins. Arginine biosynthesis and pyrimidine biosynthesis share a common intermediate, carbamoyl phosphate (CP). No pyrimidine auxotrophs were detected and only 7 L. plantarum strains required arginine for growth. Arginine auxotrophs were more frequently found in L. plantarum isolated from milk products than from fermented plant products or humans: association with dairy products might favour arginine auxotrophy. The argCJBDF genes were functional in most strains and when inactive, only one gene was mutated in more than half of the arginine auxotrophs. Random mutations may have generated these auxotrophs since different arg genes were inactivated. Analysis of the sequenced L. plantarum genome revealed the presence of 6 sets of duplicated genes in the arginine and pyrimidine biosynthetic pathways. Among the three copies of the CP synthetase large sub-unit encoding gene, pyrAb2 harboured frame-shift mutations and may be a pseudogene. These data support the hypothesis that lactic acid bacteria have adapted to specific habitats by progressively losing unnecessary genes and their genome has evolved through cumulative DNA degeneration.Lactobacillus / evolution / adaptation / mutation / auxotrophy Résumé -Évolution de certains lactobacilles par dégénérescence cumulée de l'ADN. Les bactéries lactiques nécessitent des milieux riches pour leur croissance car des mutations dans des gènes impliqués dans la biosynthèse de nombreux acides aminés, de vitamines ou de nucléobases ont invalidé leurs capacités à synthétiser ces composés. La nature et la fréquence des lésions génétiques responsables de ces auxotrophies naturelles ont été recherchées dans les gènes codant pour la synthèse de deux voies de biosynthèse, celle de l'arginine et celle des pyrimidines. Ces deux voies possèdent un intermédiaire de biosynthèse commun, le carbamyl phosphate (CP). Une collection de 150 souches de Lactobacillus plantarum isolées de produits fermentés et de niches écologiques diverses a été testée. Aucune souche n'était auxotrophe vis-à-vis des pyrimidines et 7 souches seulement pré-sentaient une auxotrophie vis-à-vis de la citrulline, un précurseur de l'arginine. Les auxotrophes Arg -étaient plus fréquemment rencontrés parmi les isolats laitiers que parmi des isolats végétaux ou d'origine humaine. Nous faisons l'hypothèse que l'association des lactobacilles aux produits laitiers favoriserait la sélection d'auxotrophes vis-à-vis de l'arginine. Les gènes impliqués dans la biosynthèse de la citrulline sont regroupés dans l'opéron argCJBDF et sont fonctionnels dans la plupart

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Growth of Lactobacillus bulgaricus in Milk. 2. Characteristics of Purine Nucleotides, Pyrimidine Nucleotides, and Nucleic Acid Synthesis

Cited by 26 publications

References 5 publications

Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium

Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium

Extent of Genetic Lesions of the Arginine and Pyrimidine Biosynthetic Pathways in Lactobacillus plantarum , L. paraplantarum, L. pentosus , and L. casei : Prevalence of CO ₂ -Dependent Auxotrophs and Characterization of Deficient arg Genes in L. plantarum

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Growth of Lactobacillus bulgaricus in Milk. 2. Characteristics of Purine Nucleotides, Pyrimidine Nucleotides, and Nucleic Acid Synthesis

Cited by 26 publications

References 5 publications

Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium

Comparative High-Density Microarray Analysis of Gene Expression during Growth of Lactobacillus helveticus in Milk versus Rich Culture Medium

Extent of Genetic Lesions of the Arginine and Pyrimidine Biosynthetic Pathways in Lactobacillus plantarum , L. paraplantarum, L. pentosus , and L. casei : Prevalence of CO 2 -Dependent Auxotrophs and Characterization of Deficient arg Genes in L. plantarum

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Extent of Genetic Lesions of the Arginine and Pyrimidine Biosynthetic Pathways in Lactobacillus plantarum , L. paraplantarum, L. pentosus , and L. casei : Prevalence of CO ₂ -Dependent Auxotrophs and Characterization of Deficient arg Genes in L. plantarum