Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum
Recombinant proteins have become useful tools in biochemistry research. During their production, however, inclusion bodies (IB) appear, on the one hand, due to the high expression rate from the recombinant plasmids, which have high efficiency promoters, and, on the other hand, intrinsic characterist...
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| Format: | Article |
| Language: | English |
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Universidad Nacional de Colombia
2017-09-01
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| Series: | Revista Colombiana de Química |
| Subjects: | |
| Online Access: | https://revistas.unal.edu.co/index.php/rcolquim/article/view/63492 |
| _version_ | 1798044027589754880 |
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| author | Carlos Alfonso Nieto Clavijo Nicolás Forero Baena María Helena Ramírez Hernández |
| author_facet | Carlos Alfonso Nieto Clavijo Nicolás Forero Baena María Helena Ramírez Hernández |
| author_sort | Carlos Alfonso Nieto Clavijo |
| collection | DOAJ |
| description | Recombinant proteins have become useful tools in biochemistry research. During their production, however, inclusion bodies (IB) appear, on the one hand, due to the high expression rate from the recombinant plasmids, which have high efficiency promoters, and, on the other hand, intrinsic characteristics of the expressed protein. Furhtermore, the nicotinamide/nicotinate mononucleotide adenilyl transferase (NMNAT) is a central protein in NAD(H)+ biosynthesis, an essential cofactor in cell metabolism, and in protozoon parasite has been studied. To study the NMNAT protein of these parasites, their recombinant version in E. coli has been expressed, getting a great quantity of IB as a by-product. To increase the solubility of the protein, the coding sequence of the NMNAT enzyme of Plasmodium falciparum was cloned in different expression plasmids which were subsequently transformed into E. coli BL21(DE3) expression strain. The solubility of the recombinant proteins was assessed and the one with the highest presence in the soluble fraction was subsequently purified and its enzyme activity was determined. The recombinant protein with a MBP (maltose-binding protein) tag showed an increased solubility and purity. |
| first_indexed | 2024-04-11T22:57:14Z |
| format | Article |
| id | doaj.art-4d11a2a318c945d7b43141c367645fad |
| institution | Directory Open Access Journal |
| issn | 0120-2804 2357-3791 |
| language | English |
| last_indexed | 2024-04-11T22:57:14Z |
| publishDate | 2017-09-01 |
| publisher | Universidad Nacional de Colombia |
| record_format | Article |
| series | Revista Colombiana de Química |
| spelling | doaj.art-4d11a2a318c945d7b43141c367645fad2022-12-22T03:58:20ZengUniversidad Nacional de ColombiaRevista Colombiana de Química0120-28042357-37912017-09-0146351010.15446/rev.colomb.quim.v46n3.6349245118Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparumCarlos Alfonso Nieto Clavijo0Nicolás Forero Baena1María Helena Ramírez Hernández2Universidad Nacional de ColombiaUniversidad Nacional de ColombiaUniversidad Nacional de ColombiaRecombinant proteins have become useful tools in biochemistry research. During their production, however, inclusion bodies (IB) appear, on the one hand, due to the high expression rate from the recombinant plasmids, which have high efficiency promoters, and, on the other hand, intrinsic characteristics of the expressed protein. Furhtermore, the nicotinamide/nicotinate mononucleotide adenilyl transferase (NMNAT) is a central protein in NAD(H)+ biosynthesis, an essential cofactor in cell metabolism, and in protozoon parasite has been studied. To study the NMNAT protein of these parasites, their recombinant version in E. coli has been expressed, getting a great quantity of IB as a by-product. To increase the solubility of the protein, the coding sequence of the NMNAT enzyme of Plasmodium falciparum was cloned in different expression plasmids which were subsequently transformed into E. coli BL21(DE3) expression strain. The solubility of the recombinant proteins was assessed and the one with the highest presence in the soluble fraction was subsequently purified and its enzyme activity was determined. The recombinant protein with a MBP (maltose-binding protein) tag showed an increased solubility and purity.https://revistas.unal.edu.co/index.php/rcolquim/article/view/63492Plasmodium falciparumsolubilidadNMNATproteína recombinantecuerpos de inclusiónTAG fusión |
| spellingShingle | Carlos Alfonso Nieto Clavijo Nicolás Forero Baena María Helena Ramírez Hernández Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum Revista Colombiana de Química Plasmodium falciparum solubilidad NMNAT proteína recombinante cuerpos de inclusión TAG fusión |
| title | Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum |
| title_full | Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum |
| title_fullStr | Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum |
| title_full_unstemmed | Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum |
| title_short | Design and production of various fusion proteins of the nicotinamide/nicotinate mononucleotide adenilil transferase (NMNAT) of Plasmodium falciparum |
| title_sort | design and production of various fusion proteins of the nicotinamide nicotinate mononucleotide adenilil transferase nmnat of plasmodium falciparum |
| topic | Plasmodium falciparum solubilidad NMNAT proteína recombinante cuerpos de inclusión TAG fusión |
| url | https://revistas.unal.edu.co/index.php/rcolquim/article/view/63492 |
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