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Characterization of Aspartate Aminotransferase Isoenzymes from Leaves of Lupinus albus L. cv Estoril
Martins, Maria Luisa Louro,Mourato, Miguel Pedro de Freitas Barbosa,Mendonca, Amarilis Paula alberti de varennes e 생화학분자생물학회 1989 BMB Reports Vol.35 No.2
Two aspartate azninotransferase (EC 2.6.1.1) isoenzymes (AAT-1 and AAT-2) from Lupines albus L. cv Estoril were separated, purified, and characterized. The molecular weight, pI value, optimum pH, optimum temperature, and thermodynamic parameters for thermal inactivation of both isaenzymes were abtained. Studies of the kinetic mechanism, and the kinetics of product inhibition and high substrate concentration inhibition, were performed. The effect of some divalent ions and irreversible inhibitors on both AAT isoenzymes was also studied. Native PAGE showed a higher molecular weight for AAT 2 compared with AAT 1. AAT 1 appears to be more anionic than AAT 2, which was suggested by the anion exchange chromatography SDS-PAGE showed a similar sub-unit molecular weight for both isoenzymes. The optimum pH (between 8.0 and 9.0) and temperature (60-65℃) were similar far both isoenzymes. In the temperature range of 45-65℃, AAT-2 has higher thermostability than AAT-1. Both isoenzymes showed a high affinity for keto-acid substrates, as well as a higher affinity to aspartate than glutamate. Manganese ions induced an increase in both AAT isoenzymes activities, but no cooperative effect was detected. Among the inhibitors tested, hydroxylamine affected both isoenzymes activity by an irreversible inhibition mechanism.
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Characterization of Aspartate Aminotransferase Isoenzymes from Leaves of Lupinus albus L. cv Estoril
Martins, Maria Luisa Louro,De Freitas Barbosa, Miguel Pedro,De Varennes E Mendonca, Amarilis Paula Alberti Korean Society for Biochemistry and Molecular Biol 2002 Journal of biochemistry and molecular biology Vol.35 No.2
Two aspartate aminoransferase (EC 2.6.1.1) isoenzymes (AAT-1 and AAT-2) from Lupinus albus L. cv Estoril were separated, purified, and characterized. The molecular weight, pI value, optimum pH, optimum temperature, and thermodynamic parameters for thermal inactivation of both isoenzymes were obtained. Studies of the kinetic mechanism, and the kinetics of product inhibition and high substrate concentration inhibition, were performed. The effect of some divalent ions and irreversible inhibitors on both AAT isoenzymes was also studied. Native PAGE showed a higher molecular weight for AAT-2 compared with AAT-1. AAT-1 appears to be more anionic than AAT-2, which was suggested by the anion exchange chromatography. SDS-PAGE showed a similar sub-unit molecular weight for both isoenzymes. The optimum pH (between 8,0 and 9.0) and temperature ($60-65^{\circ}C$) were similar for both isoenzymes. In the temperature range of $45-65^{\circ}C$, AAT-2 has higher thermostability than AAT-1. Both isoenzymes showed a high affinity for keto-acid substrates, as well as a higher affinity to aspartate than glutamate. Manganese ions induced an increase in both AAT isoenzymes activities, but no cooperative effect was detected. Among the inhibitors tested, hydroxylamine affected both isoenzymes activity by an irreversible inhibition mechanism.
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Daniela Jones-Dias,Vera Manageiro,Eugénia Ferreira,Deolinda Louro,Antibiotic Resistance Surveillance Program in Portugal Participants,Manuela Caniça 한국미생물학회 2014 The journal of microbiology Vol.52 No.6
A group of 124 Enterobacteriaceae isolates resistant to thirdgeneration cephalosporins, and collected in distinct healthcare facilities of different Portuguese regions was analysed. The great majority of the isolates were also resistant to fourthgeneration cephalosporins (83.9%), monobactam (96%),amoxicillin plus clavulanic acid (85.5%), and piperacillinplus tazobactam (66.9%). Overall, 84.7% (105/124) were multidrugresistant. Molecular methods enabled us to identify86.3% (107/124) extended-spectrum β-lactamases (ESBL)producers, revealing a diversity of class A β-lactamases fromdifferent families, like TEM (TEM-1, TEM-10, TEM-24, andTEM-52), SHV (SHV-1, SHV-12, and SHV-28), CTX-M(CTX-M-1, CTX-M-9, CTX-M-14, CTX-M-15, and CTXM-32), and GES (GES-1). We have also detected class C enzymeslike plasmid-mediated AmpC β-lactamases (PMAβs,DHA-1, and CMY-2) and chromosomal AmpCs in Enterobacterand Citrobacter spp. The PMAβ genetic context mappingsuggests association with mobile elements, plasmid importationand the potential emergence of these β-lactamases. The most prevalent β-lactamase detected was CTX-M-15(66.1%) and in 41.1% of the isolates it was associated withTEM-, OXA-type β-lactamases and Aac(6)’-Ib-cr, whichmight indicate that the respective genotype has settled inour country. Indeed, CTX-M-15 was distributed amongstdistinct clinical settings of several health care facilities(93.5%) from various regions. We provide evidence of a concerningclinical situation that includes vast occurrence ofESBLs, the settling of CTX-M β-lactamases, and the reportof plasmidic and chromosomal AmpC in Portugal.
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Mechanical Properties of Alumina Nanofilled Polymeric Composites Cured with DDSA and MNA
Ana M. Amaro,Deesy G. Pinto,Luís Bernardo,Sérgio Lopes,João Rodrigues,Cristina S. Louro 한국섬유공학회 2018 Fibers and polymers Vol.19 No.2
Reinforced concrete is widely used in structures. New materials to replace both the steel and the concrete have been studied in many research centres. One of the possibilities for the reinforcement is the partial or total replacement of the steel bars by new composite materials. Nano composites are very promising, and an investigation line was developed to this end by an interdisciplinary team. On this work, the mechanical properties of epoxy resin nanocomposites (EPNCs) filled with α-Al2O3 nanoparticles (NPs) with irregular shape and approximately 100 nm maximum diameter size was investigated. The variable study was the alumina NPs contents: 1, 3 and 5 wt.%. The NPs were previously pretreated with a silane agent (APTES). Two hardeners, 3-dodec-2-enyloxolane-2,5-dione (DDSA) and 8-methyl-3a,4,7,7a-tetrahydro-4,7-methano-2- benzofuran-1,3-dione (MNA), frequently used in epoxy resin embedding tissues, were used simultaneously for this study. Unlike other hardeners, DDSA does not need curing treatment, constituting a novel application and a saving time-energy during the manufacturing process. Considering the mechanical behaviour, it was observed that the EPNCs filled with 5 wt.% of alumina NPs showed the maximum improvement in flexural modulus, around 14 % when compared to the pristine EP sample. No relevant effect was observed on the flexural strength by adding alumina NPs. Additionally, the maximum increase observed for hardness, and Young’s modulus were about 13 % and 28 %, respectively (the maximum increase was observed at 3 wt.%).
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