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Molecular and biochemical characterization of Entamoeba histolytica fructokinase
Entamoeba histolytica is the causative agent of amoebic dysentery and liver abscess. The medium for its axenic culture contains glucose as energy source, and we addressed the question whether E. histolytica can also use fructose instead. As the amoebic hexokinases do not phosphorylate fructose, a se...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4412284/ https://www.ncbi.nlm.nih.gov/pubmed/25700717 http://dx.doi.org/10.1007/s00436-015-4383-5 |
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author | Matt, Julia Duchêne, Michael |
author_facet | Matt, Julia Duchêne, Michael |
author_sort | Matt, Julia |
collection | PubMed |
description | Entamoeba histolytica is the causative agent of amoebic dysentery and liver abscess. The medium for its axenic culture contains glucose as energy source, and we addressed the question whether E. histolytica can also use fructose instead. As the amoebic hexokinases do not phosphorylate fructose, a separate fructokinase is essential. The genome project revealed a single candidate gene encoding an E. histolytica homolog of bacterial fructokinases. This gene was cloned, and the recombinant enzyme had a magnesium-dependent fructose 6-kinase activity (EC 2.7.1.4) with a K (m) for fructose of 0.156 mM and a V (max) of 131 U/mg protein. Recombinant fructokinase also showed a much weaker mannokinase activity, but no activity with glucose or galactose. The amoebae could be switched from glucose to fructose medium without any detectable consequence on doubling time. Fructokinase messenger RNA (mRNA) was modestly but significantly upregulated in amoebae switched to fructose medium as well as in fructose-adapted E. histolytica. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00436-015-4383-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4412284 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-44122842015-05-06 Molecular and biochemical characterization of Entamoeba histolytica fructokinase Matt, Julia Duchêne, Michael Parasitol Res Original Paper Entamoeba histolytica is the causative agent of amoebic dysentery and liver abscess. The medium for its axenic culture contains glucose as energy source, and we addressed the question whether E. histolytica can also use fructose instead. As the amoebic hexokinases do not phosphorylate fructose, a separate fructokinase is essential. The genome project revealed a single candidate gene encoding an E. histolytica homolog of bacterial fructokinases. This gene was cloned, and the recombinant enzyme had a magnesium-dependent fructose 6-kinase activity (EC 2.7.1.4) with a K (m) for fructose of 0.156 mM and a V (max) of 131 U/mg protein. Recombinant fructokinase also showed a much weaker mannokinase activity, but no activity with glucose or galactose. The amoebae could be switched from glucose to fructose medium without any detectable consequence on doubling time. Fructokinase messenger RNA (mRNA) was modestly but significantly upregulated in amoebae switched to fructose medium as well as in fructose-adapted E. histolytica. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00436-015-4383-5) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2015-02-21 2015 /pmc/articles/PMC4412284/ /pubmed/25700717 http://dx.doi.org/10.1007/s00436-015-4383-5 Text en © The Author(s) 2015 https://creativecommons.org/licenses/by/4.0/ Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Paper Matt, Julia Duchêne, Michael Molecular and biochemical characterization of Entamoeba histolytica fructokinase |
title | Molecular and biochemical characterization of Entamoeba histolytica fructokinase |
title_full | Molecular and biochemical characterization of Entamoeba histolytica fructokinase |
title_fullStr | Molecular and biochemical characterization of Entamoeba histolytica fructokinase |
title_full_unstemmed | Molecular and biochemical characterization of Entamoeba histolytica fructokinase |
title_short | Molecular and biochemical characterization of Entamoeba histolytica fructokinase |
title_sort | molecular and biochemical characterization of entamoeba histolytica fructokinase |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4412284/ https://www.ncbi.nlm.nih.gov/pubmed/25700717 http://dx.doi.org/10.1007/s00436-015-4383-5 |
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