Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes

NAD(P)H oxidases (NOXs) catalyze the oxidation of NAD(P)H by reducing molecular O2 to H2O2 or H2O. Water-forming NOXs have been highlighted as promising biocatalyst systems for regeneration of NAD+ or NADP+ in industrial applications. The NADH oxidase of Giardia lamblia (GlNOX) uses naturally both n...

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Autores principales:	Adriana Castillo-Villanueva, Horacio Reyes-Vivas, Jesús Oria-Hernández
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2021
Materias:	thermal inactivation redox reactions cofactor regeneration water-forming nox Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/1e56309319d948ceb8c6f7292ea549af
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spelling	oai:doaj.org-article:1e56309319d948ceb8c6f7292ea549af2021-11-17T14:21:55ZKinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes1310-28181314-353010.1080/13102818.2021.1987325https://doaj.org/article/1e56309319d948ceb8c6f7292ea549af2021-01-01T00:00:00Zhttp://dx.doi.org/10.1080/13102818.2021.1987325https://doaj.org/toc/1310-2818https://doaj.org/toc/1314-3530NAD(P)H oxidases (NOXs) catalyze the oxidation of NAD(P)H by reducing molecular O2 to H2O2 or H2O. Water-forming NOXs have been highlighted as promising biocatalyst systems for regeneration of NAD+ or NADP+ in industrial applications. The NADH oxidase of Giardia lamblia (GlNOX) uses naturally both nicotinamide cofactors producing only water as byproduct; therefore, it can be considered a multipurpose and innocuous regeneration system for redox reactions using either NAD+ or NADP+. From a biotechnological perspective, kinetic stability (and not thermodynamic stability) must be the most important aspect to consider for enzymes involved in industrial applications; therefore, understanding the kinetic stability of a technologically interesting enzyme is an indispensable step toward its application. In this work, we analyze the thermal inactivation kinetics of GlNOX and the influence of diverse additives on it. A combination of 1,4-Dithiothreitol as reducing agent and the disaccharide trehalose protects remarkably GlNOX from inactivation. Structural studies indicate that inactivation is not related to denaturation, whereas overoxidation of the active site cysteine seems to be the main factor related to loss of the enzyme activity. It is concluded that understanding the factors that influence the protein inactivation of GlNOX allowed to improve its kinetic stability, making it biotechnologically valuable.Adriana Castillo-VillanuevaHoracio Reyes-VivasJesús Oria-HernándezTaylor & Francis Grouparticlethermal inactivationredox reactionscofactor regenerationwater-forming noxBiotechnologyTP248.13-248.65ENBiotechnology & Biotechnological Equipment, Vol 35, Iss 1, Pp 1401-1408 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	thermal inactivation redox reactions cofactor regeneration water-forming nox Biotechnology TP248.13-248.65
spellingShingle	thermal inactivation redox reactions cofactor regeneration water-forming nox Biotechnology TP248.13-248.65 Adriana Castillo-Villanueva Horacio Reyes-Vivas Jesús Oria-Hernández Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes
description	NAD(P)H oxidases (NOXs) catalyze the oxidation of NAD(P)H by reducing molecular O2 to H2O2 or H2O. Water-forming NOXs have been highlighted as promising biocatalyst systems for regeneration of NAD+ or NADP+ in industrial applications. The NADH oxidase of Giardia lamblia (GlNOX) uses naturally both nicotinamide cofactors producing only water as byproduct; therefore, it can be considered a multipurpose and innocuous regeneration system for redox reactions using either NAD+ or NADP+. From a biotechnological perspective, kinetic stability (and not thermodynamic stability) must be the most important aspect to consider for enzymes involved in industrial applications; therefore, understanding the kinetic stability of a technologically interesting enzyme is an indispensable step toward its application. In this work, we analyze the thermal inactivation kinetics of GlNOX and the influence of diverse additives on it. A combination of 1,4-Dithiothreitol as reducing agent and the disaccharide trehalose protects remarkably GlNOX from inactivation. Structural studies indicate that inactivation is not related to denaturation, whereas overoxidation of the active site cysteine seems to be the main factor related to loss of the enzyme activity. It is concluded that understanding the factors that influence the protein inactivation of GlNOX allowed to improve its kinetic stability, making it biotechnologically valuable.
format	article
author	Adriana Castillo-Villanueva Horacio Reyes-Vivas Jesús Oria-Hernández
author_facet	Adriana Castillo-Villanueva Horacio Reyes-Vivas Jesús Oria-Hernández
author_sort	Adriana Castillo-Villanueva
title	Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes
title_short	Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes
title_full	Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes
title_fullStr	Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes
title_full_unstemmed	Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes
title_sort	kinetic stability of the water-forming nadh oxidase from giardia lamblia: implications for biotechnological processes
publisher	Taylor & Francis Group
publishDate	2021
url	https://doaj.org/article/1e56309319d948ceb8c6f7292ea549af
work_keys_str_mv	AT adrianacastillovillanueva kineticstabilityofthewaterformingnadhoxidasefromgiardialambliaimplicationsforbiotechnologicalprocesses AT horacioreyesvivas kineticstabilityofthewaterformingnadhoxidasefromgiardialambliaimplicationsforbiotechnologicalprocesses AT jesusoriahernandez kineticstabilityofthewaterformingnadhoxidasefromgiardialambliaimplicationsforbiotechnologicalprocesses
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Kinetic stability of the water-forming NADH oxidase from Giardia lamblia: implications for biotechnological processes

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