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

文献类型： 外文期刊

作者： Yan, Guangbo 1 ; Li, Xia 1 ; Yang, Jun 1 ; Li, Zhongchen 1 ; Hou, Jia 1 ; Rao, Ben 2 ; Hu, Yong 3 ; Ma, Lixin 1 ; Wang, Yaping 1 ;

作者机构： 1.Hubei Univ, Fac Biol, Hubei Key Lab Ind Biotechnol, State Key Lab Biocatalysis & Enzyme,Engn Hubei Co, Wuhan 430062, Peoples R China

2.Hubei Acad Agr Sci, Biopesticide Branch Hubei Innovat Ctr Agr Sci & T, Natl Biopesticide Engn Technol Res Ctr, Hubei Biopesticide Engn Res Ctr, Wuhan 430064, Peoples R China

3.Hubei Univ Technol, Hubei Res Ctr Food Fermentat Engn & Technol, Natl Ctr Cellular Regulat & Mol Pharmaceut 111, Key Lab Fermentat Engn,Minist Educ,Hubei Key Lab, Wuhan 430062, Peoples R China

关键词： S-adenosylmethionine; ATP synthesis; scaffold protein; multi-enzyme cascade

期刊名称：BIOMOLECULES （ 影响因子：4.879； 五年影响因子：5.362 ）

ISSN：

年卷期： 2021 年 11 卷 11 期

页码：

收录情况： SCI

摘要： 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 & DEG;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 mu g of MAT4 in a reaction system with 0.3 M MgCl2 catalyzed at 20 & DEG;C and a pH of 8 catalyzed 0.5 g/L of l-Met to produce approximately 0.9 g/L of SAM. Approximately 25 mu g of MAT5 in a reaction system with 0.7 M MgCl2 catalyzed at 35 & DEG;C and a pH of 8 catalyzed 0.5 g/L of l-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.

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