Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins

Adenosine triphosphate (ATP) and S-adenosyl-L-methionine (SAM) are important intermediates that are widely present in living organisms. Large-scale preparation and application of ATP or SAM is limited by expensive raw materials. To lower the production costs for ATP/SAM, in this study we used strate...

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Autores principales:	Guangbo Yan, Xia Li, Jun Yang, Zhongchen Li, Jia Hou, Ben Rao, Yong Hu, Lixin Ma, Yaping Wang
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	S-adenosylmethionine ATP synthesis scaffold protein multi-enzyme cascade Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/d8c464f9b7444537b466a5e7fddbb861
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spelling	oai:doaj.org-article:d8c464f9b7444537b466a5e7fddbb8612021-11-25T16:54:02ZCost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins10.3390/biom111117062218-273Xhttps://doaj.org/article/d8c464f9b7444537b466a5e7fddbb8612021-11-01T00:00:00Zhttps://www.mdpi.com/2218-273X/11/11/1706https://doaj.org/toc/2218-273XAdenosine triphosphate (ATP) and S-adenosyl-L-methionine (SAM) are important intermediates that are widely present in living organisms. Large-scale preparation and application of ATP or SAM is limited by expensive raw materials. To lower the production costs for ATP/SAM, in this study we used strategies applying engineered multidomain scaffold proteins to synthesize ATP and SAM. An artificial scaffold protein containing CBM3 domain, IM proteins and CL-labeled proteins was assembled to form complex 1 for catalytic reactions to increase ATP production. The ATP synthesis system produced approximately 25 g/L of ATP with approximately 15 g/L of ADP and 5 g/L of AMP using 12.5 g/L of adenosine and 40 g/L of sodium hexametaphosphate reaction at 35 °C and a pH of 8.5 for 6 h. Based on the above ATP synthesis system, two CL-labeled methionine adenosyltransferases (CL9-MAT4 and CL9-MAT5) were applied to construct scaffold protein complex 2 to achieve SAM synthesis. Approximately 25 μg of MAT4 in a reaction system with 0.3 M MgCl<sub>2</sub> catalyzed at 20 °C and a pH of 8 catalyzed 0.5 g/L of <span style="font-variant: small-caps;">l</span>-Met to produce approximately 0.9 g/L of SAM. Approximately 25 μg of MAT5 in a reaction system with 0.7 M MgCl<sub>2</sub> catalyzed at 35 °C and a pH of 8 catalyzed 0.5 g/L of <span style="font-variant: small-caps;">l</span>-Met to produce approximately 1.2 g/L of SAM. Here, we showed that low-cost substrates can be efficiently converted into high-value additional ATP and SAM via multi-enzyme catalytic reactions by engineered multidomain scaffold proteins.Guangbo YanXia LiJun YangZhongchen LiJia HouBen RaoYong HuLixin MaYaping WangMDPI AGarticleS-adenosylmethionineATP synthesisscaffold proteinmulti-enzyme cascadeMicrobiologyQR1-502ENBiomolecules, Vol 11, Iss 1706, p 1706 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	S-adenosylmethionine ATP synthesis scaffold protein multi-enzyme cascade Microbiology QR1-502
spellingShingle	S-adenosylmethionine ATP synthesis scaffold protein multi-enzyme cascade Microbiology QR1-502 Guangbo Yan Xia Li Jun Yang Zhongchen Li Jia Hou Ben Rao Yong Hu Lixin Ma Yaping Wang Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins
description	Adenosine triphosphate (ATP) and S-adenosyl-L-methionine (SAM) are important intermediates that are widely present in living organisms. Large-scale preparation and application of ATP or SAM is limited by expensive raw materials. To lower the production costs for ATP/SAM, in this study we used strategies applying engineered multidomain scaffold proteins to synthesize ATP and SAM. An artificial scaffold protein containing CBM3 domain, IM proteins and CL-labeled proteins was assembled to form complex 1 for catalytic reactions to increase ATP production. The ATP synthesis system produced approximately 25 g/L of ATP with approximately 15 g/L of ADP and 5 g/L of AMP using 12.5 g/L of adenosine and 40 g/L of sodium hexametaphosphate reaction at 35 °C and a pH of 8.5 for 6 h. Based on the above ATP synthesis system, two CL-labeled methionine adenosyltransferases (CL9-MAT4 and CL9-MAT5) were applied to construct scaffold protein complex 2 to achieve SAM synthesis. Approximately 25 μg of MAT4 in a reaction system with 0.3 M MgCl<sub>2</sub> catalyzed at 20 °C and a pH of 8 catalyzed 0.5 g/L of <span style="font-variant: small-caps;">l</span>-Met to produce approximately 0.9 g/L of SAM. Approximately 25 μg of MAT5 in a reaction system with 0.7 M MgCl<sub>2</sub> catalyzed at 35 °C and a pH of 8 catalyzed 0.5 g/L of <span style="font-variant: small-caps;">l</span>-Met to produce approximately 1.2 g/L of SAM. Here, we showed that low-cost substrates can be efficiently converted into high-value additional ATP and SAM via multi-enzyme catalytic reactions by engineered multidomain scaffold proteins.
format	article
author	Guangbo Yan Xia Li Jun Yang Zhongchen Li Jia Hou Ben Rao Yong Hu Lixin Ma Yaping Wang
author_facet	Guangbo Yan Xia Li Jun Yang Zhongchen Li Jia Hou Ben Rao Yong Hu Lixin Ma Yaping Wang
author_sort	Guangbo Yan
title	Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins
title_short	Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins
title_full	Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins
title_fullStr	Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins
title_full_unstemmed	Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins
title_sort	cost-effective production of atp and s-adenosylmethionine using engineered multidomain scaffold proteins
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/d8c464f9b7444537b466a5e7fddbb861
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_version_	1718412865690402816

Cost-Effective Production of ATP and S-Adenosylmethionine Using Engineered Multidomain Scaffold Proteins

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