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Ahmed Fendri - One of the best experts on this subject based on the ideXlab platform.
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Functional and Structural Characterization of a Thermostable Phospholipase A2 from a Sparidae Fish (Diplodus annularis)
Journal of agricultural and food chemistry, 2017Co-Authors: Nabil Smichi, Vincent Arondel, Nabil Miled, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Youssef Gargouri, Ahmed FendriAbstract:Novel phospholipase (PLA(2)) genes from the Sparidae family were cloned. The sequenced PLA(2) revealed an identity with pancreatic PLA(2) group IB. To better understand the structure/function relationships of these enzymes and their evolution, the Diplodus annularis PLA(2) (DaPLA(2)) was overexpressed in E. coli. The refolded enzyme was purified by Ni-affinity chromatography and has a molecular mass of 15 kDa as determined by MALDI-TOF spectrometry. Interestingly, unlike the pancreatic type, the DaPIA(2) was active and stable at higher temperatures, which suggests its great potential in biotechnological applications. The 31) structure of DaPLA(2) was constructed to gain insights into the functional properties of Sparidae PLA(2). Molecular docking and dynamic simulations were performed to explain the higher thermal stability and the substrate specificity of DaPLA(2). Using the monolayer technique, the purified DaPLA(2) was found to be active on various phospholipids ranging from 10 to 20 mN-m(-1), which explained the absence of the hemolytic activity for DaPLA(2).
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Functional and Structural Characterization of a Thermostable Phospholipase A(2) from a Sparidae Fish (Diplodus annularis)
Journal of Agricultural and Food Chemistry, 2017Co-Authors: Nabil Smichi, Vincent Arondel, Nabil Miled, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Youssef Gargouri, Ahmed FendriAbstract:Novel phospholipase (PLA(2)) genes from the Sparidae family were cloned. The sequenced PLA(2) revealed an identity with pancreatic PLA(2) group IB. To better understand the structure/function relationships of these enzymes and their evolution, the Diplodus annularis PLA(2) (DaPLA(2)) was overexpressed in E. coli. The refolded enzyme was purified by Ni-affinity chromatography and has a molecular mass of 15 kDa as determined by MALDI-TOF spectrometry. Interestingly, unlike the pancreatic type, the DaPIA(2) was active and stable at higher temperatures, which suggests its great potential in biotechnological applications. The 31) structure of DaPLA(2) was constructed to gain insights into the functional properties of Sparidae PLA(2). Molecular docking and dynamic simulations were performed to explain the higher thermal stability and the substrate specificity of DaPLA(2). Using the monolayer technique, the purified DaPLA(2) was found to be active on various phospholipids ranging from 10 to 20 mN-m(-1), which explained the absence of the hemolytic activity for DaPLA(2).
Manuel A. Garrido-ramos - One of the best experts on this subject based on the ideXlab platform.
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The molecular phylogeny of the Sparidae (Pisces, Perciformes) based on two satellite DNA families.
Heredity, 2001Co-Authors: R. De La Herrán, C. Ruiz Rejón, M. Ruiz Rejón, Manuel A. Garrido-ramosAbstract:In this study, the phylogenetic relationships and which the taxonomic status of the species belonging to the Sparidae family (Pisces: Perciformes) are analysed and revised. This study includes species of this family that are distributed by the North-eastern Atlantic and Mediterranean coasts, is based on the analysis of two satellite DNA families. While one satellite DNA, the centromeric EcoRI family, extends to all the species analysed, the other, the subtelomeric DraI family, is restricted to only six of the 16 species studied. Based on phylogenetic use of these two markers, we conclude that the Sparidae family is composed by two major lineages: one comprising the species of the genera Sparus, Diplodus, Lithognathus, Boops, Sarpa and Spondyliosoma, and one species of Pagellus (P. bogaraveo); and the other lineage is comprised of the species of Pagrus and Dentex, and one species of Pagellus (P. erythrinus). This classification is consistent across the two markers used and clearly contradicts previous morphological phylogenies based mainly on dentition. In addition, the current status and the phylogenetic position of some of the species analysed (i.e. species of Pagrus, Dentex and Pagellus) are not supported by our analyses. Finally, we discuss the value of the morphological characters used until now for the classification of this group of fish.
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Evolution of centromeric satellite DNA and its use in phylogenetic studies of the Sparidae family (Pisces, Perciformes).
Molecular phylogenetics and evolution, 1999Co-Authors: Manuel A. Garrido-ramos, R. De La Herrán, Manuel Jamilena, Rafael Lozano, C. Ruiz Rejón, M. Ruiz RejónAbstract:In this paper, we use the EcoRI centromeric satellite DNA family conserved in Sparidae as a taxonomic and a phylogenetic marker. The analyses of 56 monomeric units (187 bp in size) obtained by means of cloning and PCR from 10 sparid species indicate that this repetitive DNA evolves by concerted evolution. Different phylogenetic inference methods, such as neighbor-joining and UPGMA, group the 56 repeats by taxonomic affinity and support the existence of at least two monophyletic groups within the Sparidae family. These results reinforce the recent taxonomic revision of the genera Sparus and Pagrus and contradict previous classifications of the Sparidae family.
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A satellite DNA of the Sparidae family (Pisces, Perciformes) associated with telomeric sequences
Cytogenetics and cell genetics, 1998Co-Authors: Manuel A. Garrido-ramos, R. De La Herrán, C. Ruiz Rejón, RejónAbstract:This paper reports on the isolation and localization of the subtelomeric DraI satellite DNA in the Sparidae family. Gene cloning determined that the DraI satellite DNA is present in only 3 species (Pagrus pagrus, P. auriga, and Pagellus erythrinus) of the 10 Sparidae species analyzed. The results were confirmed by PCR amplification. This satellite DNA is located in a subtelomeric position in all 48 acrocentric chromosomes of these species. However, interstitial loci are also observed. Sequence analysis of monomers of this repetitive family indicates that the satellite DNA is associated with telomeric sequences, (TTAGGG)n, in at least one species, P. erythrinus. This is the first direct demonstration of the existence of the consensus telomere sequences of vertebrates in fish. Likewise, this report also demonstrates that the ends of fish chromosomes have a structure similar to those of most eukaryote chromosomes, viz., telomere sequences and subtelomeric sequences associated by a boundary in which both types of sequences are interspersed. The recent origin of the DraI satellite DNA and its use as a phylogenetic marker is discussed.
Nabil Smichi - One of the best experts on this subject based on the ideXlab platform.
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Efficient heterologous expression, functional characterization and molecular modeling of annular seabream digestive phospholipase A2.
Chemistry and Physics of Lipids, 2017Co-Authors: Nabil Smichi, Soumaya Triki, Vincent Arondel, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Nabil MiledAbstract:Here we report the cDNA cloning of a phospholipase A2 (PLA2) from five Sparidae species. The deduced amino acid sequences show high similarity with pancreatic PLA2. In addition, a phylogenetic tree derived from alignment of various available sequences revealed that Sparidae PLA2 are closer to avian PLA2 group IB than to mammals' ones. In order to understand the structure-function relationships of these enzymes, we report here the recombinant expression in E.coli, the refolding and characterization of His-tagged annular seabream PLA2 (AsPLA2). A single Ni-affinity chromatography step was used to obtain a highly purified recombinant AsPLA2 with a molecular mass of 15kDa as attested by gel electrophoresis and MALDI-TOF mass spectrometry data. The enzyme has a specific activity of 400U.mg(-1) measured on phosphatidylcholine at pH 8.5 and 50°C. The enzyme high thermo-activity and thermo-stability make it a potential candidate in various biological applications. The 3D structure models of these enzymes were compared with structures of phylogenetically related pancreatic PLA2. By following these models and utilizing molecular dynamics simulations, the resistance of the AsPLA2 at high temperatures was explained. Using the monomolecular film technique, AsPLA2 was found to be active on various phospholipids spread at the air/water interface at a surface pressure between 12 and 25dyncm(-1). Interestingly, this enzyme was shown to be mostly active on dilauroyl-phosphatidylglycerol monolayers and this behavior was confirmed by molecular docking and dynamics simulations analysis. The discovery of a thermo-active new member of Sparidae PLA2, provides new insights on structure-activity relationships of fish PLA2.
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Functional and Structural Characterization of a Thermostable Phospholipase A2 from a Sparidae Fish (Diplodus annularis)
Journal of agricultural and food chemistry, 2017Co-Authors: Nabil Smichi, Vincent Arondel, Nabil Miled, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Youssef Gargouri, Ahmed FendriAbstract:Novel phospholipase (PLA(2)) genes from the Sparidae family were cloned. The sequenced PLA(2) revealed an identity with pancreatic PLA(2) group IB. To better understand the structure/function relationships of these enzymes and their evolution, the Diplodus annularis PLA(2) (DaPLA(2)) was overexpressed in E. coli. The refolded enzyme was purified by Ni-affinity chromatography and has a molecular mass of 15 kDa as determined by MALDI-TOF spectrometry. Interestingly, unlike the pancreatic type, the DaPIA(2) was active and stable at higher temperatures, which suggests its great potential in biotechnological applications. The 31) structure of DaPLA(2) was constructed to gain insights into the functional properties of Sparidae PLA(2). Molecular docking and dynamic simulations were performed to explain the higher thermal stability and the substrate specificity of DaPLA(2). Using the monolayer technique, the purified DaPLA(2) was found to be active on various phospholipids ranging from 10 to 20 mN-m(-1), which explained the absence of the hemolytic activity for DaPLA(2).
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Functional and Structural Characterization of a Thermostable Phospholipase A(2) from a Sparidae Fish (Diplodus annularis)
Journal of Agricultural and Food Chemistry, 2017Co-Authors: Nabil Smichi, Vincent Arondel, Nabil Miled, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Youssef Gargouri, Ahmed FendriAbstract:Novel phospholipase (PLA(2)) genes from the Sparidae family were cloned. The sequenced PLA(2) revealed an identity with pancreatic PLA(2) group IB. To better understand the structure/function relationships of these enzymes and their evolution, the Diplodus annularis PLA(2) (DaPLA(2)) was overexpressed in E. coli. The refolded enzyme was purified by Ni-affinity chromatography and has a molecular mass of 15 kDa as determined by MALDI-TOF spectrometry. Interestingly, unlike the pancreatic type, the DaPIA(2) was active and stable at higher temperatures, which suggests its great potential in biotechnological applications. The 31) structure of DaPLA(2) was constructed to gain insights into the functional properties of Sparidae PLA(2). Molecular docking and dynamic simulations were performed to explain the higher thermal stability and the substrate specificity of DaPLA(2). Using the monolayer technique, the purified DaPLA(2) was found to be active on various phospholipids ranging from 10 to 20 mN-m(-1), which explained the absence of the hemolytic activity for DaPLA(2).
M. Ruiz Rejón - One of the best experts on this subject based on the ideXlab platform.
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The molecular phylogeny of the Sparidae (Pisces, Perciformes) based on two satellite DNA families.
Heredity, 2001Co-Authors: R. De La Herrán, C. Ruiz Rejón, M. Ruiz Rejón, Manuel A. Garrido-ramosAbstract:In this study, the phylogenetic relationships and which the taxonomic status of the species belonging to the Sparidae family (Pisces: Perciformes) are analysed and revised. This study includes species of this family that are distributed by the North-eastern Atlantic and Mediterranean coasts, is based on the analysis of two satellite DNA families. While one satellite DNA, the centromeric EcoRI family, extends to all the species analysed, the other, the subtelomeric DraI family, is restricted to only six of the 16 species studied. Based on phylogenetic use of these two markers, we conclude that the Sparidae family is composed by two major lineages: one comprising the species of the genera Sparus, Diplodus, Lithognathus, Boops, Sarpa and Spondyliosoma, and one species of Pagellus (P. bogaraveo); and the other lineage is comprised of the species of Pagrus and Dentex, and one species of Pagellus (P. erythrinus). This classification is consistent across the two markers used and clearly contradicts previous morphological phylogenies based mainly on dentition. In addition, the current status and the phylogenetic position of some of the species analysed (i.e. species of Pagrus, Dentex and Pagellus) are not supported by our analyses. Finally, we discuss the value of the morphological characters used until now for the classification of this group of fish.
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Evolution of centromeric satellite DNA and its use in phylogenetic studies of the Sparidae family (Pisces, Perciformes).
Molecular phylogenetics and evolution, 1999Co-Authors: Manuel A. Garrido-ramos, R. De La Herrán, Manuel Jamilena, Rafael Lozano, C. Ruiz Rejón, M. Ruiz RejónAbstract:In this paper, we use the EcoRI centromeric satellite DNA family conserved in Sparidae as a taxonomic and a phylogenetic marker. The analyses of 56 monomeric units (187 bp in size) obtained by means of cloning and PCR from 10 sparid species indicate that this repetitive DNA evolves by concerted evolution. Different phylogenetic inference methods, such as neighbor-joining and UPGMA, group the 56 repeats by taxonomic affinity and support the existence of at least two monophyletic groups within the Sparidae family. These results reinforce the recent taxonomic revision of the genera Sparus and Pagrus and contradict previous classifications of the Sparidae family.
Nabil Miled - One of the best experts on this subject based on the ideXlab platform.
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Efficient heterologous expression, functional characterization and molecular modeling of annular seabream digestive phospholipase A2.
Chemistry and Physics of Lipids, 2017Co-Authors: Nabil Smichi, Soumaya Triki, Vincent Arondel, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Nabil MiledAbstract:Here we report the cDNA cloning of a phospholipase A2 (PLA2) from five Sparidae species. The deduced amino acid sequences show high similarity with pancreatic PLA2. In addition, a phylogenetic tree derived from alignment of various available sequences revealed that Sparidae PLA2 are closer to avian PLA2 group IB than to mammals' ones. In order to understand the structure-function relationships of these enzymes, we report here the recombinant expression in E.coli, the refolding and characterization of His-tagged annular seabream PLA2 (AsPLA2). A single Ni-affinity chromatography step was used to obtain a highly purified recombinant AsPLA2 with a molecular mass of 15kDa as attested by gel electrophoresis and MALDI-TOF mass spectrometry data. The enzyme has a specific activity of 400U.mg(-1) measured on phosphatidylcholine at pH 8.5 and 50°C. The enzyme high thermo-activity and thermo-stability make it a potential candidate in various biological applications. The 3D structure models of these enzymes were compared with structures of phylogenetically related pancreatic PLA2. By following these models and utilizing molecular dynamics simulations, the resistance of the AsPLA2 at high temperatures was explained. Using the monomolecular film technique, AsPLA2 was found to be active on various phospholipids spread at the air/water interface at a surface pressure between 12 and 25dyncm(-1). Interestingly, this enzyme was shown to be mostly active on dilauroyl-phosphatidylglycerol monolayers and this behavior was confirmed by molecular docking and dynamics simulations analysis. The discovery of a thermo-active new member of Sparidae PLA2, provides new insights on structure-activity relationships of fish PLA2.
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Functional and Structural Characterization of a Thermostable Phospholipase A2 from a Sparidae Fish (Diplodus annularis)
Journal of agricultural and food chemistry, 2017Co-Authors: Nabil Smichi, Vincent Arondel, Nabil Miled, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Youssef Gargouri, Ahmed FendriAbstract:Novel phospholipase (PLA(2)) genes from the Sparidae family were cloned. The sequenced PLA(2) revealed an identity with pancreatic PLA(2) group IB. To better understand the structure/function relationships of these enzymes and their evolution, the Diplodus annularis PLA(2) (DaPLA(2)) was overexpressed in E. coli. The refolded enzyme was purified by Ni-affinity chromatography and has a molecular mass of 15 kDa as determined by MALDI-TOF spectrometry. Interestingly, unlike the pancreatic type, the DaPIA(2) was active and stable at higher temperatures, which suggests its great potential in biotechnological applications. The 31) structure of DaPLA(2) was constructed to gain insights into the functional properties of Sparidae PLA(2). Molecular docking and dynamic simulations were performed to explain the higher thermal stability and the substrate specificity of DaPLA(2). Using the monolayer technique, the purified DaPLA(2) was found to be active on various phospholipids ranging from 10 to 20 mN-m(-1), which explained the absence of the hemolytic activity for DaPLA(2).
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Functional and Structural Characterization of a Thermostable Phospholipase A(2) from a Sparidae Fish (Diplodus annularis)
Journal of Agricultural and Food Chemistry, 2017Co-Authors: Nabil Smichi, Vincent Arondel, Nabil Miled, Houcemeddine Othman, Neila Achouri, Alexandre Noiriel, Najet Srairi-abid, Abdelkarim Abousalham, Youssef Gargouri, Ahmed FendriAbstract:Novel phospholipase (PLA(2)) genes from the Sparidae family were cloned. The sequenced PLA(2) revealed an identity with pancreatic PLA(2) group IB. To better understand the structure/function relationships of these enzymes and their evolution, the Diplodus annularis PLA(2) (DaPLA(2)) was overexpressed in E. coli. The refolded enzyme was purified by Ni-affinity chromatography and has a molecular mass of 15 kDa as determined by MALDI-TOF spectrometry. Interestingly, unlike the pancreatic type, the DaPIA(2) was active and stable at higher temperatures, which suggests its great potential in biotechnological applications. The 31) structure of DaPLA(2) was constructed to gain insights into the functional properties of Sparidae PLA(2). Molecular docking and dynamic simulations were performed to explain the higher thermal stability and the substrate specificity of DaPLA(2). Using the monolayer technique, the purified DaPLA(2) was found to be active on various phospholipids ranging from 10 to 20 mN-m(-1), which explained the absence of the hemolytic activity for DaPLA(2).
