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Updated: Jun 23, 2026
09:16
Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
Enhancing the Promiscuous Phosphorylation Activity of Acid Phosphatase via Transition-State-Stabilization-Mediated
Wei Song1,2,3, Xin Xu2,3, Wanqing Wei2,3
1School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, China.
Journal of Agricultural and Food Chemistry
|June 20, 2026
Summary
We developed a machine learning strategy (TSCF) to improve acid phosphatase enzymes for sustainable phosphorylation. This method successfully enhanced enzyme activity for producing phosphorylated compounds, demonstrating its broad applicability.
Area of Science:
- Biochemistry
- Enzyme Engineering
- Computational Biology
Background:
- Acid phosphatases catalyze phosphate ester hydrolysis.
- Some acid phosphatases exhibit phosphotransferase activity, useful for sustainable phosphorylation.
- Enzyme identification and engineering are challenging due to substrate-dependent activity variations.
Purpose of the Study:
- To develop a strategy integrating machine learning and transition-state features for enzyme mining and optimization.
- To enhance acid phosphatase-catalyzed phosphotransfer reactions for producing phosphorylated compounds.
Main Methods:
- Developed the Transition-State Conformational Feature (TSCF) strategy.
- Integrated machine learning with transition-state models and molecular dynamics simulations.
- Applied TSCF to identify and engineer acid phosphatases for specific phosphorylation reactions.
Main Results:
- TSCF showed strong predictive capability for inosine 5'-phosphate formation.
- 70.0% of designed mutants exhibited 1.2-6.4-fold higher activity than wild-type enzymes.
- Incorporating molecular dynamics improved screening accuracy to 77.8% and identified effective L-ascorbic acid phosphorylation mutants (75.0% accuracy, 71.1% conversion).
Conclusions:
- TSCF is an effective strategy for identifying and engineering acid phosphatases with improved phosphotransferase activity.
- The method demonstrates generality for enhancing enzyme-catalyzed phosphotransfer reactions.
- TSCF facilitates sustainable production of phosphorylated food-related compounds.
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