Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system

Background: The heterologous expression of parasitic proteins is challenging because the sequence composition often differs significantly from host preferences. However, the production of such proteins is important because they are potential drug targets and can be screened for interactions with new...

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Autores principales:	Julie Harnischfeger, Mandy Beutler, Denise Salzig, Stefan Rahlfs, Katja Becker, Christoph G. Grevelding, Peter Czermak
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Activity assay Aldehyde dehydrogenase Baculovirus expression vector system Drug targets Escherichia coli Metal ions Biotechnology TP248.13-248.65 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/4fd2b672fc7b402d964cdd6bbfe5e85e
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spelling	oai:doaj.org-article:4fd2b672fc7b402d964cdd6bbfe5e85e2021-11-06T04:20:17ZBiochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system0717-345810.1016/j.ejbt.2021.08.002https://doaj.org/article/4fd2b672fc7b402d964cdd6bbfe5e85e2021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S0717345821000415https://doaj.org/toc/0717-3458Background: The heterologous expression of parasitic proteins is challenging because the sequence composition often differs significantly from host preferences. However, the production of such proteins is important because they are potential drug targets and can be screened for interactions with new lead compounds. Here we compared two expression systems for the production of an active recombinant aldehyde dehydrogenase (SmALDH_312) from Schistosoma mansoni, which causes the neglected tropical disease schistosomiasis. Results: We produced SmALDH_312 successfully in the bacterium Escherichia coli and in the baculovirus expression vector system (BEVS). Both versions of the recombinant protein were found to be active in vitro, but the BEVS-derived enzyme showed 3.7-fold higher specific activity and was selected for further characterization. We investigated the influence of Mg2+, Ca2+ and Mn2+, and found out that the specific activity of the enzyme increased 1.5-fold in the presence of 0.5 mM Mg2+. Finally, we characterized the kinetic properties of the enzyme using a design-of-experiment approach, revealing optimal activity at pH 7.6 and 41°C. Conclusions: Although, E. coli has many advantages, such as rapid expression, high yields and low costs, this system was outperformed by BEVS for the production of a schistosome ALDH. BEVS therefore provides an opportunity for the expression and subsequent evaluation of schistosome enzymes as drug targets.How to cite: Harnischfeger J, Beutler M, Salzig D, et al. Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system. Electron J Biotechnol 2021;54. https://doi.org/10.1016/j.ejbt.2021.08.002Julie HarnischfegerMandy BeutlerDenise SalzigStefan RahlfsKatja BeckerChristoph G. GreveldingPeter CzermakElsevierarticleActivity assayAldehyde dehydrogenaseBaculovirus expression vector systemDrug targetsEscherichia coliMetal ionsBiotechnologyTP248.13-248.65Biology (General)QH301-705.5ENElectronic Journal of Biotechnology, Vol 54, Iss , Pp 26-36 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Activity assay Aldehyde dehydrogenase Baculovirus expression vector system Drug targets Escherichia coli Metal ions Biotechnology TP248.13-248.65 Biology (General) QH301-705.5
spellingShingle	Activity assay Aldehyde dehydrogenase Baculovirus expression vector system Drug targets Escherichia coli Metal ions Biotechnology TP248.13-248.65 Biology (General) QH301-705.5 Julie Harnischfeger Mandy Beutler Denise Salzig Stefan Rahlfs Katja Becker Christoph G. Grevelding Peter Czermak Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system
description	Background: The heterologous expression of parasitic proteins is challenging because the sequence composition often differs significantly from host preferences. However, the production of such proteins is important because they are potential drug targets and can be screened for interactions with new lead compounds. Here we compared two expression systems for the production of an active recombinant aldehyde dehydrogenase (SmALDH_312) from Schistosoma mansoni, which causes the neglected tropical disease schistosomiasis. Results: We produced SmALDH_312 successfully in the bacterium Escherichia coli and in the baculovirus expression vector system (BEVS). Both versions of the recombinant protein were found to be active in vitro, but the BEVS-derived enzyme showed 3.7-fold higher specific activity and was selected for further characterization. We investigated the influence of Mg2+, Ca2+ and Mn2+, and found out that the specific activity of the enzyme increased 1.5-fold in the presence of 0.5 mM Mg2+. Finally, we characterized the kinetic properties of the enzyme using a design-of-experiment approach, revealing optimal activity at pH 7.6 and 41°C. Conclusions: Although, E. coli has many advantages, such as rapid expression, high yields and low costs, this system was outperformed by BEVS for the production of a schistosome ALDH. BEVS therefore provides an opportunity for the expression and subsequent evaluation of schistosome enzymes as drug targets.How to cite: Harnischfeger J, Beutler M, Salzig D, et al. Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system. Electron J Biotechnol 2021;54. https://doi.org/10.1016/j.ejbt.2021.08.002
format	article
author	Julie Harnischfeger Mandy Beutler Denise Salzig Stefan Rahlfs Katja Becker Christoph G. Grevelding Peter Czermak
author_facet	Julie Harnischfeger Mandy Beutler Denise Salzig Stefan Rahlfs Katja Becker Christoph G. Grevelding Peter Czermak
author_sort	Julie Harnischfeger
title	Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system
title_short	Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system
title_full	Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system
title_fullStr	Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system
title_full_unstemmed	Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system
title_sort	biochemical characterization of the recombinant schistosome tegumental protein smaldh_312 produced in e. coli and baculovirus expression vector system
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/4fd2b672fc7b402d964cdd6bbfe5e85e
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Biochemical characterization of the recombinant schistosome tegumental protein SmALDH_312 produced in E. coli and baculovirus expression vector system

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