scholarly journals The Relationship among Tyrosine Decarboxylase and Agmatine Deiminase Pathways in Enterococcus faecalis

2017 ◽  
Vol 8 ◽  
Author(s):  
Marta Perez ◽  
Victor Ladero ◽  
Beatriz del Rio ◽  
Begoña Redruello ◽  
Anne de Jong ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Tyrosine Decarboxylase ◽  
Agmatine Deiminase ◽  
The Relationship

Heterologous Expression and Characterization of Tyrosine Decarboxylase from Enterococcus faecalis R612Z1 and Enterococcus faecium R615Z1

2014 ◽  
Vol 77 (4) ◽  
pp. 592-598 ◽  
Author(s):  
FANG LIU ◽  
WENJUAN XU ◽  
LIHUI DU ◽  
DAOYING WANG ◽  
YONGZHI ZHU ◽  
...  
Keyword(s):  
Amino Acid ◽  
Heterologous Expression ◽  
Molecular Mass ◽  
Enterococcus Faecalis ◽  
Enterococcus Faecium ◽  
Maximal Activity ◽  
Tyrosine Decarboxylase ◽  
Acid Protein ◽  
Amino Acid Protein

Tyrosine decarboxylase (TDC) is responsible for tyramine production and can catalyze phenylalanine to produce β-phenylethylamine. Enterococcus strains are a group of bacteria predominantly producing tyramine and β-phenylethylamine in water-boiled salted duck. In this study, the heterologous expression and characterization of two TDCs from Enterococcus faecalis R612Z1 (612TDC) and Enterococcus faecium R615Z1 (615TDC) were studied. The recombinant putative proteins of 612TDC and 615TDC were heterologously expressed in Escherichia coli. 612TDC is a 620-amino-acid protein with a molecular mass of 70.0 kDa, whereas 615TDC is a 625-amino-acid protein with a molecular mass of 70.3 kDa. Both 612TDC and 615TDC showed an optimum temperature of 25°C for the tyrosine and phenylalanine substrates. However, 612TDC revealed maximal activity at pH 5.5, whereas 615TDC revealed maximal activity at pH 6.0. Kinetic studies showed that 612TDC and 615TDC exhibited higher specificity for tyrosine than for phenylalanine. The catalysis abilities of both 612TDC and 615TDC for phenylalanine were restrained significantly with the increase in NaCl concentration, but this was not the case for tyrosine. This study revealed that the enzyme properties of the purified recombinant 612TDC and 615TDC were similar, although their amino acid sequences had 84% identity.

scholarly journals The Enterococcal Surface Protein, Esp, Is Involved in Enterococcus faecalis Biofilm Formation

2001 ◽  
Vol 67 (10) ◽  
pp. 4538-4545 ◽  
Author(s):  
Alejandro Toledo-Arana ◽  
Jaione Valle ◽  
Cristina Solano ◽  
Marı́a Jesús Arrizubieta ◽  
Carme Cucarella ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Biofilm Formation ◽  
Insertional Mutagenesis ◽  
Surface Protein ◽  
Structural Similarity ◽  
Abiotic Surfaces ◽  
Polyvinyl Chloride Plastic ◽  
Primary Attachment ◽  
Encoding Gene ◽  
The Relationship

ABSTRACT The enterococcal surface protein, Esp, is a high-molecular-weight surface protein of unknown function whose frequency is significantly increased among infection-derived Enterococcus faecalisisolates. In this work, a global structural similarity was found between Bap, a biofilm-associated protein of Staphylococcus aureus, and Esp. Analysis of the relationship between the presence of the Esp-encoding gene (esp) and the biofilm formation capacity in E. faecalis demonstrated that the presence of the esp gene is highly associated (P < 0.0001) with the capacity of E. faecalis to form a biofilm on a polystyrene surface, since 93.5% of the E. faecalis esp-positive isolates were capable of forming a biofilm. Moreover, none of the E. faecalis esp-deficient isolates were biofilm producers. Depending on theE. faecalis isolate, insertional mutagenesis ofesp caused either a complete loss of the biofilm formation phenotype or no apparent phenotypic defect. Complementation studies revealed that Esp expression in an E. faecalis esp-deficient strain promoted primary attachment and biofilm formation on polystyrene and polyvinyl chloride plastic from urine collection bags. Together, these results demonstrate that (i) biofilm formation capacity is widespread among clinical E. faecalis isolates, (ii) the biofilm formation capacity is restricted to the E. faecalis strains harboringesp, and (iii) Esp promotes primary attachment and biofilm formation of E. faecalis on abiotic surfaces.

scholarly journals The Gene Cluster for Agmatine Catabolism of Enterococcus faecalis: Study of Recombinant Putrescine Transcarbamylase and Agmatine Deiminase and a Snapshot of Agmatine Deiminase Catalyzing Its Reaction

2006 ◽  
Vol 189 (4) ◽  
pp. 1254-1265 ◽  
Author(s):  
José L. Llácer ◽  
Luis Mariano Polo ◽  
Sandra Tavárez ◽  
Benito Alarcón ◽  
Rebeca Hilario ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Enterococcus Faecalis ◽  
Duplication Event ◽  
Arginine Deiminase ◽  
Substrate Affinity ◽  
En Bloc ◽  
Carbamate Kinase ◽  
Atp Generation ◽  
Agmatine Deiminase ◽  
Covalent Catalysis

ABSTRACT Enterococcus faecalis makes ATP from agmatine in three steps catalyzed by agmatine deiminase (AgDI), putrescine transcarbamylase (PTC), and carbamate kinase (CK). An antiporter exchanges putrescine for agmatine. We have cloned the E. faecalis ef0732 and ef0734 genes of the reported gene cluster for agmatine catabolism, overexpressed them in Escherichia coli, purified the products, characterized them functionally as PTC and AgDI, and crystallized and X-ray diffracted them. The 1.65-Å-resolution structure of AgDI forming a covalent adduct with an agmatine-derived amidine reactional intermediate is described. We provide definitive identification of the gene cluster for agmatine catabolism and confirm that ornithine is a genuine but poor PTC substrate, suggesting that PTC (found here to be trimeric) evolved from ornithine transcarbamylase. N-(Phosphonoacetyl)-putrescine was prepared and shown to strongly (Ki = 10 nM) and selectively inhibit PTC and to improve PTC crystallization. We find that E. faecalis AgDI, which is committed to ATP generation, closely resembles the AgDIs involved in making polyamines, suggesting the recruitment of a polyamine-synthesizing AgDI into the AgDI pathway. The arginine deiminase (ADI) pathway of arginine catabolism probably supplied the genes for PTC and CK but not those for the agmatine/putrescine antiporter, and thus the AgDI and ADI pathways are not related by a single “en bloc” duplication event. The AgDI crystal structure reveals a tetramer with a five-blade propeller subunit fold, proves that AgDI closely resembles ADI despite a lack of sequence identity, and explains substrate affinity, selectivity, and Cys357-mediated-covalent catalysis. A three-tongued agmatine-triggered gating opens or blocks access to the active center.

scholarly journals Expression of the Agmatine Deiminase Pathway in Enterococcus faecalis Is Activated by the AguR Regulator and Repressed by CcpA and PTSMan Systems

PLoS ONE ◽  
2013 ◽  
Vol 8 (10) ◽  
pp. e76170 ◽  
Author(s):  
Cristian Suárez ◽  
Martín Espariz ◽  
Víctor S. Blancato ◽  
Christian Magni
Keyword(s):  
Enterococcus Faecalis ◽  
Agmatine Deiminase

scholarly journals Replication of Enterococcus faecalis Pheromone-Responding Plasmid pAD1: Location of the Minimal Replicon and oriV Site and RepA Involvement in Initiation of Replication

2004 ◽  
Vol 186 (15) ◽  
pp. 5003-5016 ◽  
Author(s):  
Maria Victoria Francia ◽  
Shuhei Fujimoto ◽  
Patricia Tille ◽  
Keith E. Weaver ◽  
Don B. Clewell
Keyword(s):  
Enterococcus Faecalis ◽  
Copy Number ◽  
Replication Origin ◽  
Plasmid Vector ◽  
Small Segment ◽  
Initiator Protein ◽  
Repeat Sequences ◽  
Initiation Of Replication ◽  
Relationship Of ◽  
The Relationship

ABSTRACT The hemolysin-determining plasmid pAD1 is a member of a widely disseminated family of highly conjugative elements commonly present in clinical isolates of Enterococcus faecalis. The determinants repA, repB, and repC, as well as adjacent iteron sequences, are believed to play important roles in pAD1 replication and maintenance. The repA gene encodes an initiator protein, whereas repB and repC encode proteins related to stability and copy number. The present study focuses specifically on repA and identifies a replication origin (oriV) within a central region of the repA determinant. A small segment of repA carrying oriV was able to support replication in cis of a plasmid vector otherwise unable to replicate, if an intact RepA was supplied in trans. We demonstrate that under conditions in which RepA is expressed from an artificial promoter, a segment of DNA carrying only repA is sufficient for stable replication in E. faecalis. We also show that RepA binds specifically to oriV DNA at several sites containing inverted repeat sequences (i.e., IR-1) and nonspecifically to single-stranded DNA, and related genetic analyses confirm that these sequences play an important role in replication. Finally, we reveal a relationship between the internal structure of RepA and its ability to recognize oriV. An in-frame deletion within repA resulting in loss of 105 nucleotides, including at least part of oriV, did not eliminate the ability of the altered RepA protein to initiate replication using an intact origin provided in trans. The relationship of RepA to other known initiator proteins is also discussed.

Production of Tyramine by Enterococcus faecalis Strains in Water-Boiled Salted Duck

2013 ◽  
Vol 76 (5) ◽  
pp. 854-859 ◽  
Author(s):  
FANG LIU ◽  
LIHUI DU ◽  
WEIYAN XU ◽  
DAOYING WANG ◽  
MUHAN ZHANG ◽  
...  
Keyword(s):  
Amino Acids ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Biogenic Amines ◽  
Enterococcus Faecalis ◽  
High Performance ◽  
Tyrosine Decarboxylase ◽  
Pcr Assays ◽  
De Man ◽  
Layer Chromatography

The potential to produce biogenic amines was investigated with 15 Lactococcus lactis and 15 Enterococcus faecalis strains isolated from water-boiled salted duck. The production of biogenic amines from the isolated strains grown in de Man Rogosa Sharpe broth containing precursor amino acids was determined by thin-layer chromatography and high-performance liquid chromatography. None of the L. lactis strains produced any biogenic amines, whereas 12 strains of E. faecalis produced tyramine and β-phenylethylamine. PCR assays were used to detect the presence of tyrosine decarboxylase genes in all of the isolated strains. Only the 12 biogenic amine–producing Enterococcus strains had a 924-bp fragment characteristic for the tyrosine decarboxylase gene. The comparison of the amplified partial tyrDC gene sequences of the 12 positive Enterococcus strains revealed 99% similarity within the same species. The tyramine production of the sterilized water-boiled salted duck inoculated with E. faecalis R612Z1 increased significantly during storage. This study reveals that the isolated E. faecalis strains can produce tyramine and β-phenylethylamine in the medium; however, they can only produce tyramine in water-boiled salted duck.

Effect of NaCl Treatments on Tyramine Biosynthesis of Enterococcus faecalis

2015 ◽  
Vol 78 (5) ◽  
pp. 940-945 ◽  
Author(s):  
FANG LIU ◽  
XINXIN WANG ◽  
LIHUI DU ◽  
DAOYING WANG ◽  
YONGZHI ZHU ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Genetic Regulation ◽  
Nacl Stress ◽  
Negative Regulator ◽  
Logarithmic Phase ◽  
Relative Gene Expression ◽  
Tyrosine Decarboxylase ◽  
Culture Time ◽  
Nacl Concentration ◽  
Relative Gene

The effect of NaCl stress (0 to 8%, wt/vol) on the growth and tyramine production in two Enterococcus faecalis strains was examined during culture time. The growth of E. faecalis was inhibited by the increase in NaCl concentration, but tyramine production was unaffected. Tyramine accumulated rapidly during the logarithmic phase of the strains, and the final tyramine levels were approximately 800 μg/ml. Relative gene expression of four genes in the tyrosine decarboxylase locus, namely, tyrRS, tyrDC, tyrP, and nhaC, was evaluated at different incubation times. The results showed that NaCl stress could upregulate the expression of tyrDC and tyrP to improve the tyramine production of a single E. faecalis strain under certain conditions, and TyrS could act as a negative regulator on the genetic regulation of the tyramine cluster.

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