Bibliographic citations
Vargas, J., (2019). Clonación y caracterización funcional de la enzima L-Galactosa deshidrogenasa de la vía D-Manosa/L-Galactosa de Myrciaria dubia (Kunth) McVaugh [Tesis, Universidad Nacional de la Amazonía Peruana]. http://repositorio.unapiquitos.edu.pe/handle/20.500.12737/6591
Vargas, J., Clonación y caracterización funcional de la enzima L-Galactosa deshidrogenasa de la vía D-Manosa/L-Galactosa de Myrciaria dubia (Kunth) McVaugh [Tesis]. : Universidad Nacional de la Amazonía Peruana; 2019. http://repositorio.unapiquitos.edu.pe/handle/20.500.12737/6591
@misc{renati/970853,
title = "Clonación y caracterización funcional de la enzima L-Galactosa deshidrogenasa de la vía D-Manosa/L-Galactosa de Myrciaria dubia (Kunth) McVaugh",
author = "Vargas Santillan, Jhon Antoni",
publisher = "Universidad Nacional de la Amazonía Peruana",
year = "2019"
}
In the Peruvian Amazon, the fruits of Myrciaria dubia are mainly characterized by their high content of ascorbic acid (AA), which represents an alternative for the development of biotechnological processes of AA production. In camucamu, for the biosynthesis of AA there are 5 metabolic pathways, however, none of these pathways has been characterized. Here, we report the study of the enzyme L-galactose dehydrogenase (GDH) of the D-mannose / Lgalactose pathway of M. dubia, that was cloned and functionally characterized. For this, the sequence of the gene encoding the enzyme L-galactose dehydrogenase was identified in the M. dubia transcriptome, the sequence was optimized and insert into the expression vector (pET-TOPO), this was expressed in Escherichia coli Rosetta BL21 (DE3). The enzyme was purified by affinity and molecular exclusion chromatography, eluting a molecule with a calculated molecular mass of 38.6 kDa (theoretical mass 39.3 kDa), demonstrating that the enzyme is a monomer in solution. The recombinant enzyme L-galactose dehydrogenase of M. dubia (MdGDH) showed activity against L-galactose in the presence of NAD +, likewise, it showed a Km= 0.3533 effect of a Michaelis-Menten kinetics. The results obtained suggest that L-galactose dehydrogenase would be related to the high content of AA in camu-camu, likewise, this work provides the model to perform studies of the other enzymes that are involved in the biosynthesis of AA.
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