The Experts below are selected from a list of 240 Experts worldwide ranked by ideXlab platform
Andras Miczak - One of the best experts on this subject based on the ideXlab platform.
-
Recombinant Mycobacterium Smegmatis vaccine candidates.
Acta Microbiologica Et Immunologica Hungarica, 2011Co-Authors: Ildiko Faludi, Ágnes Míra Szabó, Katalin Burián, Valéria Endrész, Andras MiczakAbstract:Mycobacterium Smegmatis is a species of rapidly growing saprophytes with a number of properties that make it an effective vaccine vector. Recombinant M. Smegmatis expressing protective antigens of different pathogens and molecules modulating the immune responses offers some potential for reduction of the burden of tuberculosis, HIV and hepatitis B infections. This paper discusses the molecular methods used to generate recombinant M. Smegmatis and the results obtained with some of these recombinants.
-
MSMEG_4626 ribonuclease from Mycobacterium Smegmatis
Molecular Biology Reports, 2009Co-Authors: Agnes Csanadi, Ildiko Faludi, Andras MiczakAbstract:The MSMEG_4626 gene was cloned from Mycobacterium Smegmatis MC2 155. It codes for a protein of 1,037 amino acids, identified as ribonuclease E by matching to the protein family HMM TIGR00757. The protein was expressed and purified. Although its calculated molecular weight is 112.7 kDa, it has an aberrant mobility in SDS-polyacrylamide gels, like other ribonuclease E enzymes (it migrates as a 180 kDa protein). The central part of the protein displays high similarity to the catalytic domains of other RNase E enzymes. Mass spectrometric analysis revealed the presence of the chaperonin GroEL, ribosomal proteins, a negative regulator of genetic competence and GTP pyrophosphokinase in the affinity-purified preparation. It is a very unstable protein; despite the use of protease inhibitors in addition to the full-length RNase E its proteolytic fragments were detected.
Madanodaya Sundhoro - One of the best experts on this subject based on the ideXlab platform.
-
selective targeting of Mycobacterium Smegmatis with trehalose functionalized nanoparticles
Chemical Communications, 2015Co-Authors: Kalana W Jayawardana, Surangi H N Jayawardena, Samurdhi A Wijesundera, Thareendra De Zoysa, Madanodaya SundhoroAbstract:Silica and iron oxide nanoparticles with sizes ranging from 6 to 40 nm were functionalized with trehalose. The trehalose-conjugated nanoparticles showed strong interactions with Mycobacterium Smegmatis (M. Smegmatis) and minimal interactions with macrophage (RAW 264.7) or A549 cells. In addition, trehalose-conjugated silica nanoparticles selectively interacted with M. Smegmatis on M. Smegmatis-treated A549 cells, demonstrating high potential of trehalose in developing targeted therapy for treating mycobacterial infection.
Ildiko Faludi - One of the best experts on this subject based on the ideXlab platform.
-
Recombinant Mycobacterium Smegmatis vaccine candidates.
Acta Microbiologica Et Immunologica Hungarica, 2011Co-Authors: Ildiko Faludi, Ágnes Míra Szabó, Katalin Burián, Valéria Endrész, Andras MiczakAbstract:Mycobacterium Smegmatis is a species of rapidly growing saprophytes with a number of properties that make it an effective vaccine vector. Recombinant M. Smegmatis expressing protective antigens of different pathogens and molecules modulating the immune responses offers some potential for reduction of the burden of tuberculosis, HIV and hepatitis B infections. This paper discusses the molecular methods used to generate recombinant M. Smegmatis and the results obtained with some of these recombinants.
-
MSMEG_4626 ribonuclease from Mycobacterium Smegmatis
Molecular Biology Reports, 2009Co-Authors: Agnes Csanadi, Ildiko Faludi, Andras MiczakAbstract:The MSMEG_4626 gene was cloned from Mycobacterium Smegmatis MC2 155. It codes for a protein of 1,037 amino acids, identified as ribonuclease E by matching to the protein family HMM TIGR00757. The protein was expressed and purified. Although its calculated molecular weight is 112.7 kDa, it has an aberrant mobility in SDS-polyacrylamide gels, like other ribonuclease E enzymes (it migrates as a 180 kDa protein). The central part of the protein displays high similarity to the catalytic domains of other RNase E enzymes. Mass spectrometric analysis revealed the presence of the chaperonin GroEL, ribosomal proteins, a negative regulator of genetic competence and GTP pyrophosphokinase in the affinity-purified preparation. It is a very unstable protein; despite the use of protease inhibitors in addition to the full-length RNase E its proteolytic fragments were detected.
Surangi H N Jayawardena - One of the best experts on this subject based on the ideXlab platform.
-
selective targeting of Mycobacterium Smegmatis with trehalose functionalized nanoparticles
Chemical Communications, 2015Co-Authors: Kalana W Jayawardana, Surangi H N Jayawardena, Samurdhi A Wijesundera, Thareendra De Zoysa, Madanodaya SundhoroAbstract:Silica and iron oxide nanoparticles with sizes ranging from 6 to 40 nm were functionalized with trehalose. The trehalose-conjugated nanoparticles showed strong interactions with Mycobacterium Smegmatis (M. Smegmatis) and minimal interactions with macrophage (RAW 264.7) or A549 cells. In addition, trehalose-conjugated silica nanoparticles selectively interacted with M. Smegmatis on M. Smegmatis-treated A549 cells, demonstrating high potential of trehalose in developing targeted therapy for treating mycobacterial infection.
Abbouni Bouziane - One of the best experts on this subject based on the ideXlab platform.
-
Development of Rapid Assay for Ribonucleotide Reduction by Mycobacterium Smegmatis Mc 2 155 and their Biochemical
2020Co-Authors: Bendaha Mohammed Lamine, Abbouni BouzianeAbstract:Mycobacterium Smegmatis mc 2 155 contains a Ribonucleotide reductases (RNR), which catalyses the irreversible reduction of ribonucleotides to the corresponding 2´deoxyribonucleotides required for DNA replication and cell proliferation. 2 155 were permeabilized with two organic solvents toluene and ether for two times (2,5 min) to develop a new assay for ribonucleotide reduction. Due the importance of the growth phase in determining the yield of biomass and ribonucleotide reductase activity of Mycobacterium Smegmatis mc 2 155, a correlation between Ribonucleotide reductase activity and growth of Mycobacterium Smegmatis mc2 155 in modified seed medium has been investigated. For the enrichment of the Ribonucleotide reductase, different purification procedure has been achieved by using fast protein liquid chromatography (FPLC) with superdex G-200 chromatography and Phenyl-Superose HR 5/5 and the enzyme activity was assayed by using (HPLC). Ribonucleotide reductase activity was detectable in the 40-60% ammonium sulphate fraction. A further purification procedure by gel filtration on the superdex G-200 led to a dissociation of the both subunits. Therefore, a biochemical complementation assay was necessary to identify ribonucleotide reductase activity. The obtained specific activity of the purified protein was 1790 pmol per mg per min with an overall yield of 10%. The purified small subunit of MS2- protein was detected on SDS-PAGE, which was showed a strong band that corresponds to an apparent molecular weight of 38.5 KDa. The obtained results of ribonuleotide reduction activity with ether permeabilized cells of Mycobacterium Smegmatis mc 2 155 presented a comparable enzyme activity for both times, while with toluene permeabilized cells indicated a low enzyme activity. Furthermore, the obtained results of the correlation between ribonucleotide reductase activity and the growth showed that Ribonucleotide redutase is a peak enzyme. Finally, the permeabilisation of the cells of M. Smegmatis mc 2 155 with ether and toluene for short time facilitated us to develop a rapid assay for ribonucleotide reductase activity of others gram positive bacteria.
-
Development of Rapid Assay for Ribonucleotide Reduction by Mycobacterium Smegmatis Mc2 155 and their Biochemical Characterisation
Journal of biotechnology & biomaterials, 2012Co-Authors: Elhachemi Ahmed, Bendaha Mohammed Lamine, Benattouche Zouaoui, Kanoun Khedoudja, Abbouni BouzianeAbstract:Mycobacterium Smegmatis mc2 155 contains a Ribonucleotide reductases (RNR), which catalyses the irreversible reduction of ribonucleotides to the corresponding 2 Ideoxyribonucleotides required for DNA replication and cell proliferation. The aim of this work was the development of a rapid assay for ribonucleotide reduction by Mycobacterium Smegmatis mc 2 155 and their biochemical characterisation. For this purpose, the cells of Mycobacterium Smegmatis mc 2 155 were permeabilized with two organic solvents toluene and ether for two times (2,5 min) to develop a new assay for ribonucleotide reduction. Due the importance of the growth phase in determining the yield of biomass and ribonucleotide reductase activity of Mycobacterium Smegmatis mc 2 155, a correlation between Ribonucleotide reductase activity and growth of Mycobacterium Smegmatis mc 2 155 in modified seed medium has been investigated. For the enrichment of the Ribonucleotide reductase, different purification procedure has been achieved by using fast protein liquid chromatography (FPLC) with superdex G-200 chromatography and Phenyl-Superose HR 5/5 and the enzyme activity was assayed by using (HPLC). Ribonucleotide reductase activity was detectable in the 40-60% ammonium sulphate fraction. A further purification procedure by gel filtration on the superdex G-200 led to a dissociation of the both subunits. Therefore, a biochemical complementation assay was necessary to identify ribonucleotide reductase activity. The obtained specific activity of the purified protein was 1790 pmol per mg per min with an overall yield of 10%. The purified small subunit of MS2- protein was detected on SDS-PAGE, which was showed a strong band that corresponds to an apparent molecular weight of 38.5 KDa. The obtained results of ribonuleotide reduction activity with ether permeabilized cells of Mycobacterium Smegmatis mc 2 155 presented a comparable enzyme activity for both times, while with toluene permeabilized cells indicated a low enzyme activity. Furthermore, the obtained results of the correlation between ribonucleotide reductase activity and the growth showed that Ribonucleotide redutase is a peak enzyme. Finally, the permeabilisation of the cells of M. Smegmatis mc 2 155 with ether and toluene for short time facilitated us to develop a rapid assay for ribonucleotide reductase activity of others gram positive bacteria.
