In vitro biosynthesis of ATP from adenosine and polyphosphate

Abstract Adenosine triphosphate (ATP) acts as a crucial energy currency in vivo, and it is a widely used energy and/or phosphate donor for enzyme-catalyzed reactions in vitro. In this study, we established an in vitro multi-enzyme cascade system for ATP production. Using adenosine and inorganic poly...

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Autores principales:	Chuanqi Sun, Zonglin Li, Xiao Ning, Wentian Xu, Zhimin Li
Formato:	article
Lenguaje:	EN
Publicado:	SpringerOpen 2021
Materias:	Adenosine triphosphate In vitro Multi-enzymatic cascade catalysis Polyphosphate kinase Adenosine kinase Technology T Chemical technology TP1-1185 Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/9e691c0ee07f414bac67f16c584c750b
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spelling	oai:doaj.org-article:9e691c0ee07f414bac67f16c584c750b2021-12-05T12:03:57ZIn vitro biosynthesis of ATP from adenosine and polyphosphate10.1186/s40643-021-00469-02197-4365https://doaj.org/article/9e691c0ee07f414bac67f16c584c750b2021-11-01T00:00:00Zhttps://doi.org/10.1186/s40643-021-00469-0https://doaj.org/toc/2197-4365Abstract Adenosine triphosphate (ATP) acts as a crucial energy currency in vivo, and it is a widely used energy and/or phosphate donor for enzyme-catalyzed reactions in vitro. In this study, we established an in vitro multi-enzyme cascade system for ATP production. Using adenosine and inorganic polyphosphate (polyP) as key substrates, we combined adenosine kinase and two functionally distinct polyphosphate kinases (PPKs) in a one-pot reaction to achieve chain-like ATP regeneration and production. Several sources of PPK were screened and characterized, and two suitable PPKs were selected to achieve high rates of ATP production. Among these, Sulfurovum lithotrophicum PPK (SlPPK) exhibited excellent activity over a wide pH range (pH 4.0–9.0) and synthesized ATP from ADP using short-chain polyP. Furthermore, it had a half-life > 155.6 h at 45 °C. After optimizing the reaction conditions, we finally carried out the coupling-catalyzed reaction with different initial adenosine concentrations of 10, 20, and 30 mM. The highest yields of ATP were 76.0, 70.5, and 61.3%, respectively. Graphical AbstractChuanqi SunZonglin LiXiao NingWentian XuZhimin LiSpringerOpenarticleAdenosine triphosphateIn vitroMulti-enzymatic cascade catalysisPolyphosphate kinaseAdenosine kinaseTechnologyTChemical technologyTP1-1185BiotechnologyTP248.13-248.65ENBioresources and Bioprocessing, Vol 8, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Adenosine triphosphate In vitro Multi-enzymatic cascade catalysis Polyphosphate kinase Adenosine kinase Technology T Chemical technology TP1-1185 Biotechnology TP248.13-248.65
spellingShingle	Adenosine triphosphate In vitro Multi-enzymatic cascade catalysis Polyphosphate kinase Adenosine kinase Technology T Chemical technology TP1-1185 Biotechnology TP248.13-248.65 Chuanqi Sun Zonglin Li Xiao Ning Wentian Xu Zhimin Li In vitro biosynthesis of ATP from adenosine and polyphosphate
description	Abstract Adenosine triphosphate (ATP) acts as a crucial energy currency in vivo, and it is a widely used energy and/or phosphate donor for enzyme-catalyzed reactions in vitro. In this study, we established an in vitro multi-enzyme cascade system for ATP production. Using adenosine and inorganic polyphosphate (polyP) as key substrates, we combined adenosine kinase and two functionally distinct polyphosphate kinases (PPKs) in a one-pot reaction to achieve chain-like ATP regeneration and production. Several sources of PPK were screened and characterized, and two suitable PPKs were selected to achieve high rates of ATP production. Among these, Sulfurovum lithotrophicum PPK (SlPPK) exhibited excellent activity over a wide pH range (pH 4.0–9.0) and synthesized ATP from ADP using short-chain polyP. Furthermore, it had a half-life > 155.6 h at 45 °C. After optimizing the reaction conditions, we finally carried out the coupling-catalyzed reaction with different initial adenosine concentrations of 10, 20, and 30 mM. The highest yields of ATP were 76.0, 70.5, and 61.3%, respectively. Graphical Abstract
format	article
author	Chuanqi Sun Zonglin Li Xiao Ning Wentian Xu Zhimin Li
author_facet	Chuanqi Sun Zonglin Li Xiao Ning Wentian Xu Zhimin Li
author_sort	Chuanqi Sun
title	In vitro biosynthesis of ATP from adenosine and polyphosphate
title_short	In vitro biosynthesis of ATP from adenosine and polyphosphate
title_full	In vitro biosynthesis of ATP from adenosine and polyphosphate
title_fullStr	In vitro biosynthesis of ATP from adenosine and polyphosphate
title_full_unstemmed	In vitro biosynthesis of ATP from adenosine and polyphosphate
title_sort	in vitro biosynthesis of atp from adenosine and polyphosphate
publisher	SpringerOpen
publishDate	2021
url	https://doaj.org/article/9e691c0ee07f414bac67f16c584c750b
work_keys_str_mv	AT chuanqisun invitrobiosynthesisofatpfromadenosineandpolyphosphate AT zonglinli invitrobiosynthesisofatpfromadenosineandpolyphosphate AT xiaoning invitrobiosynthesisofatpfromadenosineandpolyphosphate AT wentianxu invitrobiosynthesisofatpfromadenosineandpolyphosphate AT zhiminli invitrobiosynthesisofatpfromadenosineandpolyphosphate
_version_	1718372273645158400

In vitro biosynthesis of ATP from adenosine and polyphosphate

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