標題: Mass spectrometry-guided refinement of chemical energy buffers
作者: Chen, T. -R.
Urban, P. L.
應用化學系
應用化學系分子科學碩博班
Department of Applied Chemistry
Institute of Molecular science
關鍵字: adenosine triphosphate;biocatalysis;biosynthesis;enzyme;mass spectrometry;nucleotide recycling
公開日期: 1-Jun-2016
摘要: Biocatalytic reactions often require supplying chemical energy and phosphate groups in the form of adenosine triphosphate (ATP). Auxiliary enzymes can be used to convert a reaction by-product-adenosine diphosphate (ADP)-back to ATP. By employing real-time mass spectrometry (RTMS), one can gain an insight into inter-conversions of reactants in multi-enzyme reaction systems and optimize the reaction conditions. In this study, temporal traces of ions corresponding to adenosine monophosphate (AMP), ADP and ATP provided vital information that could be used to adjust activities of the \'\'buffering enzymes\'. Using the RTMS results as a feedback, we also characterized a bienzymatic energy buffer that enables the recovery of ATP in the cases where it is directly hydrolysed to AMP in the main enzymatic reaction. The significance of careful selection of enzyme activities-guided by RTMS-is exemplified in the synthesis of glucose-6-phosphate by hexokinase in the presence of a buffering enzyme, pyruvate kinase. Relative activities of the two enzymes, present in the reaction mixture, influence biosynthetic reaction yields. This observation supports the conclusion that optimization of chemical energy recycling procedures is critical for the biosynthetic reaction economy.
URI: http://dx.doi.org/10.1098/rspa.2015.0812
http://hdl.handle.net/11536/133941
ISSN: 1364-5021
DOI: 10.1098/rspa.2015.0812
期刊: PROCEEDINGS OF THE ROYAL SOCIETY A-MATHEMATICAL PHYSICAL AND ENGINEERING SCIENCES
Volume: 472
Issue: 2190
Appears in Collections:Articles