Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies

Abstract Catalytically active inclusion bodies (CatIBs) produced in Escherichia coli are an interesting but currently underexplored strategy for enzyme immobilization. They can be purified easily and used directly as stable and reusable heterogenous catalysts. However, very few examples of CatIBs th...

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Autores principales:	Sarah Kamel, Miriam C. Walczak, Felix Kaspar, Sarah Westarp, Peter Neubauer, Anke Kurreck
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/07f86537fc4c442f8b39ad9c8c5b8c25
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spelling	oai:doaj.org-article:07f86537fc4c442f8b39ad9c8c5b8c252021-12-02T15:10:54ZThermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies10.1038/s41598-021-96073-52045-2322https://doaj.org/article/07f86537fc4c442f8b39ad9c8c5b8c252021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96073-5https://doaj.org/toc/2045-2322Abstract Catalytically active inclusion bodies (CatIBs) produced in Escherichia coli are an interesting but currently underexplored strategy for enzyme immobilization. They can be purified easily and used directly as stable and reusable heterogenous catalysts. However, very few examples of CatIBs that are naturally formed during heterologous expression have been reported so far. Previous studies have revealed that the adenosine 5′-monophosphate phosphorylase of Thermococcus kodakarensis (TkAMPpase) forms large soluble multimers with high thermal stability. Herein, we show that heat treatment of soluble protein from crude extract induces aggregation of active protein which phosphorolyse all natural 5′-mononucleotides. Additionally, inclusion bodies formed during the expression in E. coli were found to be similarly active with 2–6 folds higher specific activity compared to these heat-induced aggregates. Interestingly, differences in the substrate preference were observed. These results show that the recombinant thermostable TkAMPpase is one of rare examples of naturally formed CatIBs.Sarah KamelMiriam C. WalczakFelix KasparSarah WestarpPeter NeubauerAnke KurreckNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sarah Kamel Miriam C. Walczak Felix Kaspar Sarah Westarp Peter Neubauer Anke Kurreck Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies
description	Abstract Catalytically active inclusion bodies (CatIBs) produced in Escherichia coli are an interesting but currently underexplored strategy for enzyme immobilization. They can be purified easily and used directly as stable and reusable heterogenous catalysts. However, very few examples of CatIBs that are naturally formed during heterologous expression have been reported so far. Previous studies have revealed that the adenosine 5′-monophosphate phosphorylase of Thermococcus kodakarensis (TkAMPpase) forms large soluble multimers with high thermal stability. Herein, we show that heat treatment of soluble protein from crude extract induces aggregation of active protein which phosphorolyse all natural 5′-mononucleotides. Additionally, inclusion bodies formed during the expression in E. coli were found to be similarly active with 2–6 folds higher specific activity compared to these heat-induced aggregates. Interestingly, differences in the substrate preference were observed. These results show that the recombinant thermostable TkAMPpase is one of rare examples of naturally formed CatIBs.
format	article
author	Sarah Kamel Miriam C. Walczak Felix Kaspar Sarah Westarp Peter Neubauer Anke Kurreck
author_facet	Sarah Kamel Miriam C. Walczak Felix Kaspar Sarah Westarp Peter Neubauer Anke Kurreck
author_sort	Sarah Kamel
title	Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies
title_short	Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies
title_full	Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies
title_fullStr	Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies
title_full_unstemmed	Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies
title_sort	thermostable adenosine 5′-monophosphate phosphorylase from thermococcus kodakarensis forms catalytically active inclusion bodies
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/07f86537fc4c442f8b39ad9c8c5b8c25
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Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies

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