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Hideyuki Suzuki - One of the best experts on this subject based on the ideXlab platform.
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a novel Putrescine exporter sapbcdf of escherichia coli
Journal of Biological Chemistry, 2016Co-Authors: Yuta Sugiyama, Kyohei Higashi, Kazuei Igarashi, Atsuo Nakamura, Mitsuharu Matsumoto, Ayaka Kanbe, Mikiyasu Sakanaka, Takane Katayama, Hideyuki SuzukiAbstract:Recent research has suggested that polyamines (Putrescine, spermidine, and spermine) in the intestinal tract impact the health of animals either negatively or positively. The concentration of polyamines in the intestinal tract results from the balance of uptake and export of the intestinal bacteria. However, the mechanism of polyamine export from bacterial cells to the intestinal lumen is still unclear. In Escherichia coli, PotE was previously identified as a transporter responsible for Putrescine excretion in an acidic growth environment. We observed Putrescine concentration in the culture supernatant was increased from 0 to 50 μm during growth of E. coli under neutral conditions. Screening for the unidentified Putrescine exporter was performed using a gene knock-out collection of E. coli, and deletion of sapBCDF significantly decreased Putrescine levels in the culture supernatant. Complementation of the deletion mutant with the sapBCDF genes restored Putrescine levels in the culture supernatant. Additionally, the ΔsapBCDF strain did not facilitate uptake of Putrescine from the culture supernatant. Quantification of stable isotope-labeled Putrescine derived from stable isotope-labeled arginine supplemented in the medium revealed that SapBCDF exported Putrescine from E. coli cells to the culture supernatant. It was previously reported that SapABCDF of Salmonella enterica sv. typhimurium and Haemophilus influenzae conferred resistance toantimicrobial peptides; however, the E. coli ΔsapBCDF strain did not affect resistance to antimicrobial peptide LL-37. These results strongly suggest that the natural function of the SapBCDF proteins is the export of Putrescine.
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Putrescine importer plap contributes to swarming motility and urothelial cell invasion in proteus mirabilis
Journal of Biological Chemistry, 2013Co-Authors: Shi Kurihara, Yumi Sakai, Aaro Muth, Otto Phanstiel, Hideyuki Suzuki, Philip N RatheAbstract:Previously, we reported that the speA gene, encoding arginine decarboxylase, is required for swarming in the urinary tract pathogen Proteus mirabilis. In addition, this previous study suggested that Putrescine may act as a cell-to-cell signaling molecule (Sturgill, G., and Rather, P. N. (2004) Mol. Microbiol. 51, 437–446). In this new study, PlaP, a putative Putrescine importer, was characterized in P. mirabilis. In a wild-type background, a plaP null mutation resulted in a modest swarming defect and slightly decreased levels of intracellular Putrescine. In a P. mirabilis speA mutant with greatly reduced levels of intracellular Putrescine, plaP was required for the Putrescine-dependent rescue of swarming motility. When a speA/plaP double mutant was grown in the presence of extracellular Putrescine, the intracellular levels of Putrescine were greatly reduced compared with the speA mutant alone, indicating that PlaP functioned as the primary Putrescine importer. In urothelial cell invasion assays, a speA mutant exhibited a 50% reduction in invasion when compared with wild type, and this defect could be restored by Putrescine in a PlaP-dependent manner. The Putrescine analog Triamide-44 partially inhibited the uptake of Putrescine by PlaP and decreased both Putrescine stimulated swarming and urothelial cell invasion in a speA mutant.
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a novel Putrescine importer required for type 1 pili driven surface motility induced by extracellular Putrescine in escherichia coli k 12
Journal of Biological Chemistry, 2011Co-Authors: Shin Kurihara, Hideyuki Suzuki, Mayu Oshida, Yoshimi BennoAbstract:Recently, many studies have reported that polyamines play a role in bacterial cell-to-cell signaling processes. The present study describes a novel Putrescine importer required for induction of type 1 pili-driven surface motility. The surface motility of the Escherichia coli ΔspeAB ΔspeC ΔpotABCD strain, which cannot produce Putrescine and cannot import spermidine from the medium, was induced by extracellular Putrescine. Introduction of the gene deletions for known polyamine importers (ΔpotE, ΔpotFGHI, and ΔpuuP) or a putative polyamine importer (ΔydcSTUV) into the ΔspeAB ΔspeC ΔpotABCD strain did not affect Putrescine-induced surface motility. The deletion of yeeF, an annotated putative Putrescine importer, in the ΔspeAB ΔspeC ΔpotABCD ΔydcSTUV strain abolished surface motility in Putrescine-supplemented medium. Complementation of yeeF by a plasmid vector restored surface motility. The surface motility observed in the present study was abolished by the deletion of fimA, suggesting that the surface motility is type 1 pili-driven. A transport assay using the yeeF+ or ΔyeeF strains revealed that YeeF is a novel Putrescine importer. The Km of YeeF (155 μm) is 40 to 300 times higher than that of other importers reported previously. On the other hand, the Vmax of YeeF (9.3 nmol/min/mg) is comparable to that of PotABCD, PotFGHI, and PuuP. The low affinity of YeeF for Putrescine may allow E. coli to sense the cell density depending on the concentration of extracellular Putrescine.
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a Putrescine inducible pathway comprising puue ynei in which γ aminobutyrate is degraded into succinate in escherichia coli k 12
Journal of Bacteriology, 2010Co-Authors: Shin Kurihara, Hidehiko Kumagai, Kenji Kato, Kei Asada, Hideyuki SuzukiAbstract:γ-Aminobutyrate (GABA) is metabolized to succinic semialdehyde by GABA aminotransferase (GABA-AT), and the succinic semialdehyde is subsequently oxidized to succinate by succinic semialdehyde dehydrogenase (SSADH). In Escherichia coli, there are duplicate GABA-ATs (GabT and PuuE) and duplicate SSADHs (GabD and YneI). While GabT and GabD have been well studied previously, the characterization and expression analysis of PuuE and YneI are yet to be investigated. By analyzing the amino acid profiles in cells of ΔpuuE and/or ΔgabT mutants, this study demonstrated that PuuE plays an important role in GABA metabolism in E. coli cells. The similarity of the amino acid sequences of PuuE and GabT is 67.4%, and it was biochemically demonstrated that the catalytic center of GabT is conserved as an amino acid residue important for the enzymatic activity in PuuE as Lys-247. However, the regulation of expression of PuuE is significantly different from that of GabT. PuuE is induced by the addition of Putrescine to the medium and is repressed by succinate and low aeration conditions; in contrast, GabT is almost constitutive. Similarly, YneI is induced by Putrescine, while GabD is not. For E. coli, PuuE is important for utilization of Putrescine as a sole nitrogen source and both PuuE and YneI are important for utilization of Putrescine as a sole carbon source. The results demonstrate that the PuuE-YneI pathway was a Putrescine-inducible GABA degradation pathway for utilizing Putrescine as a nutrient source.
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the Putrescine importer puup of escherichia coli k 12
Journal of Bacteriology, 2009Co-Authors: Shin Kurihara, Shinpei Oda, Yuichi Tsuboi, Hyeon Guk Kim, Hidehiko Kumagai, Hideyuki SuzukiAbstract:The Puu pathway is a Putrescine utilization pathway involving gamma-glutamyl intermediates. The genes encoding the enzymes of the Puu pathway form a gene cluster, the puu gene cluster, and puuP is one of the genes in this cluster. In Escherichia coli, three Putrescine importers, PotFGHI, PotABCD, and PotE, were discovered in the 1990s and have been studied; however, PuuP had not been discovered previously. This paper shows that PuuP is a novel Putrescine importer whose kinetic parameters are equivalent to those of the polyamine importers discovered previously. A puuP+ strain absorbed up to 5 mM Putrescine from the medium, but a ΔpuuP strain did not. E. coli strain MA261 has been used in previous studies of polyamine transporters, but PuuP had not been identified previously. It was revealed that the puuP gene of MA261 was inactivated by a point mutation. When E. coli was grown on minimal medium supplemented with Putrescine as the sole carbon or nitrogen source, only PuuP among the polyamine importers was required. puuP was expressed strongly when Putrescine was added to the medium or when the puuR gene, which encodes a putative repressor, was deleted. When E. coli was grown in M9-tryptone medium, PuuP was expressed mainly in the exponential growth phase, and PotFGHI was expressed independently of the growth phase.
Shin Kurihara - One of the best experts on this subject based on the ideXlab platform.
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functional identification of Putrescine c and n hydroxylases
ACS Chemical Biology, 2016Co-Authors: Tiffany M Lowepower, Shin Kurihara, Stephen S Gonzales, Jacinth Naidoo, John B Macmillan, Caitilyn Allen, Anthony J MichaelAbstract:The small polyamine Putrescine (1,4-diaminobutane) is ubiquitously and abundantly found in all three domains of life. It is a precursor, through N-aminopropylation or N-aminobutylation, for biosynthesis of the longer polyamines spermidine, sym-homospermidine, spermine, and thermospermine and longer and branched chain polyamines. Putrescine is also biochemically modified for purposes of metabolic regulation and catabolism, e.g. N-acetylation and N-glutamylation, and for incorporation into specialized metabolites, e.g. N-methylation, N-citrylation, N-palmitoylation, N-hydroxylation, and N-hydroxycinnamoylation. Only one example is known where Putrescine is modified on a methylene carbon: the formation of 2-hydroxyPutrescine by an unknown C-hydroxylase. Here, we report the functional identification of a previously undescribed Putrescine 2-hydroxylase, a Rieske-type nonheme iron sulfur protein from the β-proteobacteria Bordetella bronchiseptica and Ralstonia solanacearum. Identification of the Putrescine 2-hy...
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Functional Identification of Putrescine C- and N‑Hydroxylases
2016Co-Authors: Tiffany Lowe-power, Shin Kurihara, Jacinth Naidoo, John B Macmillan, Caitilyn Allen, Stephen Gonzales, Anthony J MichaelAbstract:The small polyamine Putrescine (1,4-diaminobutane) is ubiquitously and abundantly found in all three domains of life. It is a precursor, through N-aminopropylation or N-aminobutylation, for biosynthesis of the longer polyamines spermidine, sym-homospermidine, spermine, and thermospermine and longer and branched chain polyamines. Putrescine is also biochemically modified for purposes of metabolic regulation and catabolism, e.g. N-acetylation and N-glutamylation, and for incorporation into specialized metabolites, e.g. N-methylation, N-citrylation, N-palmitoylation, N-hydroxylation, and N-hydroxycinnamoylation. Only one example is known where Putrescine is modified on a methylene carbon: the formation of 2-hydroxyPutrescine by an unknown C-hydroxylase. Here, we report the functional identification of a previously undescribed Putrescine 2-hydroxylase, a Rieske-type nonheme iron sulfur protein from the β-proteobacteria Bordetella bronchiseptica and Ralstonia solanacearum. Identification of the Putrescine 2-hydroxylase will facilitate investigation of the physiological functions of 2-hydroxyPutrescine. One known role of 2-hydroxyPutrescine has direct biomedical relevance: its role in the biosynthesis of the cyclic hydroxamate siderophore alcaligin, a potential virulence factor of the causative agent of whooping cough, Bordetella pertussis. We also report the functional identification of a Putrescine N-hydroxylase from the γ-proteobacterium Shewanella oneidensis, which is homologous to FAD- and NADPH-dependent ornithine and lysine N-monooxygenases involved in siderophore biosynthesis. Heterologous expression of the Putrescine N-hydroxylase in E. coli produced free N-hydroxyPutrescine, never detected previously in a biological system. Furthermore, the Putrescine C- and N-hydroxylases identified here could contribute new functionality to polyamine structural scaffolds, including C–H bond functionalization in synthetic biology strategies
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a novel Putrescine importer required for type 1 pili driven surface motility induced by extracellular Putrescine in escherichia coli k 12
Journal of Biological Chemistry, 2011Co-Authors: Shin Kurihara, Hideyuki Suzuki, Mayu Oshida, Yoshimi BennoAbstract:Recently, many studies have reported that polyamines play a role in bacterial cell-to-cell signaling processes. The present study describes a novel Putrescine importer required for induction of type 1 pili-driven surface motility. The surface motility of the Escherichia coli ΔspeAB ΔspeC ΔpotABCD strain, which cannot produce Putrescine and cannot import spermidine from the medium, was induced by extracellular Putrescine. Introduction of the gene deletions for known polyamine importers (ΔpotE, ΔpotFGHI, and ΔpuuP) or a putative polyamine importer (ΔydcSTUV) into the ΔspeAB ΔspeC ΔpotABCD strain did not affect Putrescine-induced surface motility. The deletion of yeeF, an annotated putative Putrescine importer, in the ΔspeAB ΔspeC ΔpotABCD ΔydcSTUV strain abolished surface motility in Putrescine-supplemented medium. Complementation of yeeF by a plasmid vector restored surface motility. The surface motility observed in the present study was abolished by the deletion of fimA, suggesting that the surface motility is type 1 pili-driven. A transport assay using the yeeF+ or ΔyeeF strains revealed that YeeF is a novel Putrescine importer. The Km of YeeF (155 μm) is 40 to 300 times higher than that of other importers reported previously. On the other hand, the Vmax of YeeF (9.3 nmol/min/mg) is comparable to that of PotABCD, PotFGHI, and PuuP. The low affinity of YeeF for Putrescine may allow E. coli to sense the cell density depending on the concentration of extracellular Putrescine.
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a Putrescine inducible pathway comprising puue ynei in which γ aminobutyrate is degraded into succinate in escherichia coli k 12
Journal of Bacteriology, 2010Co-Authors: Shin Kurihara, Hidehiko Kumagai, Kenji Kato, Kei Asada, Hideyuki SuzukiAbstract:γ-Aminobutyrate (GABA) is metabolized to succinic semialdehyde by GABA aminotransferase (GABA-AT), and the succinic semialdehyde is subsequently oxidized to succinate by succinic semialdehyde dehydrogenase (SSADH). In Escherichia coli, there are duplicate GABA-ATs (GabT and PuuE) and duplicate SSADHs (GabD and YneI). While GabT and GabD have been well studied previously, the characterization and expression analysis of PuuE and YneI are yet to be investigated. By analyzing the amino acid profiles in cells of ΔpuuE and/or ΔgabT mutants, this study demonstrated that PuuE plays an important role in GABA metabolism in E. coli cells. The similarity of the amino acid sequences of PuuE and GabT is 67.4%, and it was biochemically demonstrated that the catalytic center of GabT is conserved as an amino acid residue important for the enzymatic activity in PuuE as Lys-247. However, the regulation of expression of PuuE is significantly different from that of GabT. PuuE is induced by the addition of Putrescine to the medium and is repressed by succinate and low aeration conditions; in contrast, GabT is almost constitutive. Similarly, YneI is induced by Putrescine, while GabD is not. For E. coli, PuuE is important for utilization of Putrescine as a sole nitrogen source and both PuuE and YneI are important for utilization of Putrescine as a sole carbon source. The results demonstrate that the PuuE-YneI pathway was a Putrescine-inducible GABA degradation pathway for utilizing Putrescine as a nutrient source.
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the Putrescine importer puup of escherichia coli k 12
Journal of Bacteriology, 2009Co-Authors: Shin Kurihara, Shinpei Oda, Yuichi Tsuboi, Hyeon Guk Kim, Hidehiko Kumagai, Hideyuki SuzukiAbstract:The Puu pathway is a Putrescine utilization pathway involving gamma-glutamyl intermediates. The genes encoding the enzymes of the Puu pathway form a gene cluster, the puu gene cluster, and puuP is one of the genes in this cluster. In Escherichia coli, three Putrescine importers, PotFGHI, PotABCD, and PotE, were discovered in the 1990s and have been studied; however, PuuP had not been discovered previously. This paper shows that PuuP is a novel Putrescine importer whose kinetic parameters are equivalent to those of the polyamine importers discovered previously. A puuP+ strain absorbed up to 5 mM Putrescine from the medium, but a ΔpuuP strain did not. E. coli strain MA261 has been used in previous studies of polyamine transporters, but PuuP had not been identified previously. It was revealed that the puuP gene of MA261 was inactivated by a point mutation. When E. coli was grown on minimal medium supplemented with Putrescine as the sole carbon or nitrogen source, only PuuP among the polyamine importers was required. puuP was expressed strongly when Putrescine was added to the medium or when the puuR gene, which encodes a putative repressor, was deleted. When E. coli was grown in M9-tryptone medium, PuuP was expressed mainly in the exponential growth phase, and PotFGHI was expressed independently of the growth phase.
Miguel A. Alvarez - One of the best experts on this subject based on the ideXlab platform.
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the biogenic amines Putrescine and cadaverine show in vitro cytotoxicity at concentrations that can be found in foods
Scientific Reports, 2019Co-Authors: Beatriz Del Rio, Daniel M Linares, Victor Ladero, Begona Redruello, Maria Fernandez, Patricia Ruasmadiedo, Cruz M Martin, Miguel A. AlvarezAbstract:Putrescine and cadaverine are among the most common biogenic amines (BA) in foods, but it is advisable that their accumulation be avoided. Present knowledge about their toxicity is, however, limited; further research is needed if qualitative and quantitative risk assessments for foods are to be conducted. The present work describes a real-time analysis of the cytotoxicity of Putrescine and cadaverine on intestinal cell cultures. Both BA were cytotoxic at concentrations found in BA-rich foods, although the cytotoxicity threshold for cadaverine was twice that of Putrescine. Their mode of cytotoxic action was similar, with both BA causing cell necrosis; they did not induce apoptosis. The present results may help in establishing legal limits for both Putrescine and cadaverine in food.
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Lactobacillus rossiae strain isolated from sourdough produces Putrescine from arginine
Nature Publishing Group, 2018Co-Authors: Beatriz Del Rio, Victor Ladero, Begona Redruello, Maria Cruz Martin, Maria Fernandez, Patricia Alvarez-sieiro, Miguel A. AlvarezAbstract:Abstract This work reports a Lactobacillus rossiae strain (L. rossiae D87) isolated from sourdough that synthesizes Putrescine - a biogenic amine that raises food safety and spoilage concerns - from arginine via the ornithine decarboxylase (ODC) pathway. The odc and potE genes were identified and sequenced. These genes respectively encode ornithine decarboxylase (Odc), which participates in the decarboxylation of ornithine to Putrescine, and the ornithine/Putrescine exchanger (PotE), which exchanges ornithine for Putrescine. Transcriptional analysis showed that odc and potE form an operon that is regulated transcriptionally by ornithine in a dose-dependent manner. To explore the possible role of the ODC pathway as an acid stress resistance mechanism for this bacterium, the effect of acidic pHs on its transcriptional regulation and on Putrescine biosynthesis was analysed. Acidic pHs induced the transcription of the odc-potE genes and the production of Putrescine over that seen at neutral pH. Further, Putrescine production via the ODC system improved the survival of L. rossiae D87 by counteracting the acidification of the cytoplasm when the cells were subjected to acidic conditions. These results suggest the ODC pathway of L. rossiae D87 provides a biochemical defence mechanism against acidic environments
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the relationship among tyrosine decarboxylase and agmatine deiminase pathways in enterococcus faecalis
Frontiers in Microbiology, 2017Co-Authors: Marta Perez, Victor Ladero, Begona Redruello, Maria Fernandez, Beatriz Del Rio, Cruz M Martin, Anne De Jong, Oscar P Kuipers, Jan Kok, Miguel A. AlvarezAbstract:Enterococci are considered mainly responsible for the undesirable accumulation of the biogenic amines (BA) tyramine and Putrescine in cheeses. The biosynthesis of tyramine and Putrescine has been described as a species trait in Enterococcus faecalis. Tyramine is formed by the decarboxylation of the amino acid tyrosine, by the tyrosine decarboxylase (TDC) route encoded in the tdc cluster. Putrescine is formed from agmatine by the agmatine deiminase (AGDI) pathway encoded in the agdi cluster. These biosynthesis routes have been independently studied, tyrosine and agmatine transcriptionally regulate the tdc and agdi clusters. The objective of the present work is to study the possible co-regulation among TDC and AGDI pathways in E. faecalis. In the presence of agmatine, a positive correlation between Putrescine biosynthesis and the tyrosine concentration was found. Transcriptome studies showed that tyrosine induces the transcription of Putrescine biosynthesis genes and up-regulates pathways involved in cell growth. The tyrosine modulation over AGDI route was not observed in the mutant Δtdc strain. Fluorescence analyses using gfp as reporter protein revealed PaguB (the promoter of agdi catabolic genes) was induced by tyrosine in the wild-type but not in the mutant strain, confirming that tdc cluster was involved in the tyrosine induction of Putrescine biosynthesis. This study also suggests that AguR (the transcriptional regulator of agdi) was implicated in interaction among the two clusters.
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lactose mediated carbon catabolite repression of Putrescine production in dairy lactococcus lactis is strain dependent
Food Microbiology, 2015Co-Authors: Beatriz Del Rio, Daniel M Linares, Victor Ladero, Begona Redruello, Maria Cruz Martin, Maria Fernandez, Miguel A. AlvarezAbstract:Lactococcus lactis is the lactic acid bacterial (LAB) species most widely used as a primary starter in the dairy industry. However, several strains of L. lactis produce the biogenic amine Putrescine via the agmatine deiminase (AGDI) pathway. We previously reported the Putrescine biosynthesis pathway in L. lactis subsp. cremoris GE2-14 to be regulated by carbon catabolic repression (CCR) via glucose but not lactose (Linares et al., 2013). The present study shows that both these sugars repress Putrescine biosynthesis in L. lactis subsp. lactis T3/33, a strain isolated from a Spanish artisanal cheese. Furthermore, we demonstrated that both glucose and lactose repressed the transcriptional activity of the aguBDAC catabolic genes of the AGDI route. Finally, a screening performed in Putrescine-producing dairy L. lactis strains determined that Putrescine biosynthesis was repressed by lactose in all the L. lactis subsp. lactis strains tested, but in only one L. lactis subsp. cremoris strain. Given the obvious importance of the lactose-repression in cheese Putrescine accumulation, it is advisable to consider the diversity of L. lactis in this sense and characterize consequently the starter cultures to select the safest strains.
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Putrescine production via the agmatine deiminase pathway increases the growth of lactococcus lactis and causes the alkalinization of the culture medium
Applied Microbiology and Biotechnology, 2015Co-Authors: Daniel M Linares, Victor Ladero, Begona Redruello, Maria Cruz Martin, Maria Fernandez, Miguel A. AlvarezAbstract:Lactococcus lactis is the most important starter culture organism used in the dairy industry. Although L. lactis species have been awarded Qualified Presumption of Safety status by the European Food Safety Authority, and Generally Regarded as Safe status by the US Food and Drug Administration, some strains can produce the biogenic amine Putrescine. One such strain is L. lactis subsp. cremoris CECT 8666 (formerly L. lactis subsp. cremoris GE2-14), which was isolated from Genestoso cheese. This strain catabolizes agmatine to Putrescine via the agmatine deiminase (AGDI) pathway, which involves the production of ATP and two ammonium ions. The present work shows that the availability of agmatine and its metabolization to Putrescine allows for greater bacterial growth (in a biphasic pattern) and causes the alkalinization of the culture medium in a dose-dependent manner. The construction of a mutant lacking the AGDI cluster (L. lactis CECT 8666 Δagdi) confirmed the latter’s direct role in Putrescine production, growth, and medium alkalinization. Alkalinization did not affect the Putrescine production pattern and was not essential for increased bacterial growth.
Hidehiko Kumagai - One of the best experts on this subject based on the ideXlab platform.
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a Putrescine inducible pathway comprising puue ynei in which γ aminobutyrate is degraded into succinate in escherichia coli k 12
Journal of Bacteriology, 2010Co-Authors: Shin Kurihara, Hidehiko Kumagai, Kenji Kato, Kei Asada, Hideyuki SuzukiAbstract:γ-Aminobutyrate (GABA) is metabolized to succinic semialdehyde by GABA aminotransferase (GABA-AT), and the succinic semialdehyde is subsequently oxidized to succinate by succinic semialdehyde dehydrogenase (SSADH). In Escherichia coli, there are duplicate GABA-ATs (GabT and PuuE) and duplicate SSADHs (GabD and YneI). While GabT and GabD have been well studied previously, the characterization and expression analysis of PuuE and YneI are yet to be investigated. By analyzing the amino acid profiles in cells of ΔpuuE and/or ΔgabT mutants, this study demonstrated that PuuE plays an important role in GABA metabolism in E. coli cells. The similarity of the amino acid sequences of PuuE and GabT is 67.4%, and it was biochemically demonstrated that the catalytic center of GabT is conserved as an amino acid residue important for the enzymatic activity in PuuE as Lys-247. However, the regulation of expression of PuuE is significantly different from that of GabT. PuuE is induced by the addition of Putrescine to the medium and is repressed by succinate and low aeration conditions; in contrast, GabT is almost constitutive. Similarly, YneI is induced by Putrescine, while GabD is not. For E. coli, PuuE is important for utilization of Putrescine as a sole nitrogen source and both PuuE and YneI are important for utilization of Putrescine as a sole carbon source. The results demonstrate that the PuuE-YneI pathway was a Putrescine-inducible GABA degradation pathway for utilizing Putrescine as a nutrient source.
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the Putrescine importer puup of escherichia coli k 12
Journal of Bacteriology, 2009Co-Authors: Shin Kurihara, Shinpei Oda, Yuichi Tsuboi, Hyeon Guk Kim, Hidehiko Kumagai, Hideyuki SuzukiAbstract:The Puu pathway is a Putrescine utilization pathway involving gamma-glutamyl intermediates. The genes encoding the enzymes of the Puu pathway form a gene cluster, the puu gene cluster, and puuP is one of the genes in this cluster. In Escherichia coli, three Putrescine importers, PotFGHI, PotABCD, and PotE, were discovered in the 1990s and have been studied; however, PuuP had not been discovered previously. This paper shows that PuuP is a novel Putrescine importer whose kinetic parameters are equivalent to those of the polyamine importers discovered previously. A puuP+ strain absorbed up to 5 mM Putrescine from the medium, but a ΔpuuP strain did not. E. coli strain MA261 has been used in previous studies of polyamine transporters, but PuuP had not been identified previously. It was revealed that the puuP gene of MA261 was inactivated by a point mutation. When E. coli was grown on minimal medium supplemented with Putrescine as the sole carbon or nitrogen source, only PuuP among the polyamine importers was required. puuP was expressed strongly when Putrescine was added to the medium or when the puuR gene, which encodes a putative repressor, was deleted. When E. coli was grown in M9-tryptone medium, PuuP was expressed mainly in the exponential growth phase, and PotFGHI was expressed independently of the growth phase.
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γ glutamylPutrescine synthetase in the Putrescine utilization pathway of escherichia coli k 12
Journal of Biological Chemistry, 2008Co-Authors: Shin Kurihara, Shinpei Oda, Yuichi Tsuboi, Hyeon Guk Kim, Mayu Oshida, Hidehiko Kumagai, Hideyuki SuzukiAbstract:Glutamate-Putrescine ligase (gamma-glutamylPutrescine synthetase, PuuA, EC 6.3.1.11) catalyzes the gamma-glutamylation of Putrescine, the first step in a novel Putrescine utilization pathway involving gamma-glutamylated intermediates, the Puu pathway, in Escherichia coli. In this report, the character and physiological importance of PuuA are described. Purified non-tagged PuuA catalyzed the ATP-dependent gamma-glutamylation of Putrescine. The K(m) values for glutamate, ATP, and Putrescine are 2.07, 2.35, and 44.6 mm, respectively. There are two Putrescine utilization pathways in E. coli: the Puu pathway and the pathway without gamma-glutamylation. Gene deletion experiments of puuA, however, indicated that the Puu pathway was more critical in utilizing Putrescine as a sole carbon or nitrogen source. The transcription of puuA was induced by Putrescine and in a puuR-deleted strain. The amino acid sequences of PuuA and glutamine synthetase (GS) show high similarity. The molecular weights of the monomers of the two enzymes are quite similar, and PuuA exists as a dodecamer as does GS. Moreover the two amino acid residues of E. coli GS that are important for the metal-catalyzed oxidation of the enzyme molecule involved in protein turnover are conserved in PuuA, and it was experimentally shown that the corresponding amino acid residues in PuuA were involved in the metal-catalyzed oxidation similarly to GS. It is suggested that the intracellular concentration of Putrescine is optimized by PuuA transcriptionally and posttranslationally and that excess Putrescine is converted to a nutrient source by the Puu pathway.
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a novel Putrescine utilization pathway involves γ glutamylated intermediates of escherichia coli k 12
Journal of Biological Chemistry, 2005Co-Authors: Shin Kurihara, Shinpei Oda, Hyeon Guk Kim, Hidehiko Kumagai, Kenji Kato, Takashi Koyanagi, Hideyuki SuzukiAbstract:A novel bacterial Putrescine utilization pathway was discovered. Seven genes, the functions of whose products were not known, are involved in this novel pathway. Five of them encode enzymes that catabolize Putrescine; one encodes a Putrescine importer, and the other encodes a transcriptional regulator. This novel pathway involves six sequential steps as follows: 1) import of Putrescine; 2) ATP-dependent gamma-glutamylation of Putrescine; 3) oxidization of gamma-glutamylPutrescine; 4) dehydrogenation of gamma-glutamyl-gamma-aminobutyraldehyde; 5) hydrolysis of the gamma-glutamyl linkage of gamma-glutamyl-gamma-aminobutyrate; and 6) transamination of gamma-aminobutyrate to form the final product of this pathway, succinate semialdehyde, which is the precursor of succinate.
Isabel Pardo - One of the best experts on this subject based on the ideXlab platform.
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ability of kocuria varians lth 1540 to degrade Putrescine identification and characterization of a novel amine oxidase
Journal of Agricultural and Food Chemistry, 2015Co-Authors: Sara Callejon, Ramon Sendra, Sergi Ferrer, Isabel PardoAbstract:This work describes the identification and characterization of an amine oxidase from Kocuria varians LTH 1540 (syn. Micrococcus varians) primarily acting on Putrescine. Data from MALDI-TOF MS/MS and the identification of Δ1-pyrroline as degradation product from Putrescine indicate that the enzyme is a flavin-dependent Putrescine oxidase (PuO). Properties of partially purified enzyme have been determined. The enzyme oxidizes diamines, Putrescine and cadaverine, and, to a lesser extent, polyamines, such as spermidine, but not monoamines. The kinetic constants (Km and Vmax) for the two major substrates were 94 ± 10 μM and 2.3 ± 0.1 μmol/min·mg for Putrescine and 75 ± 5 μM and 0.15 ± 0.02 μmol/min·mg for cadaverine. Optimal temperature and pH were 45 °C and 8.5, respectively. Enzyme was stable until 50 °C. K. varians PuO is sensitive to human flavin-dependent amine oxidase inhibitors and carboxyl-modifying compounds. The new enzyme has been isolated from a bacterial starter used in the manufacture of fermente...
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factors affecting the production of Putrescine from agmatine by lactobacillus hilgardii x1b isolated from wine
Journal of Applied Microbiology, 2008Co-Authors: M E Arena, Jose Maria Landete, M Manca C De Nadra, Isabel PardoAbstract:Aims: To elucidate and characterize the metabolic Putrescine synthesis pathway from agmatine by Lactobacillus hilgardii X1B. Methods and Results: The Putrescine formation from agmatine by resting cells (the normal physiological state in wine) of lactic acid bacteria isolated from wine has been determined for the first time. Agmatine deiminase and N-carbamoylPutrescine hydrolase enzymes, determined by HPLC and LC-Ion Trap Mass Spectrometry, carried out the Putrescine synthesis from agmatine. The influence of pH, temperature, organic acids, amino acids, sugars and ethanol on the Putrescine formation in wine was determined. Conclusions: Resting cells of Lact. hilgardii X1B produce Putrescine in wine. The Putrescine production was carried out from agmatine through the agmatine deiminase system. Significance and Impact of the Study: These results have significance from two points of view, wine quality and toxicological and microbiological aspects, taking account that Putrescine, which origin is still controversial, is quantitatively the main biogenic amine found in wine.
