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Updated: Aug 6, 2026
09:27
Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues
Published on: February 17, 2017
Construction of a Self-Assembled Multi-Enzyme Cascade for Efficient D-Allulose Biosynthesis
Zhe-Ming Wu1,2,3, Yu Lu3, Jin-Chao Zhang3
1State Key Laboratory of Green Chemical Synthesis and Conversion, Zhejiang University of Technology, Hangzhou, China.
Biotechnology and Bioengineering
|July 26, 2026
Summary
Researchers developed a peptide-mediated assembly strategy to improve d-allulose biosynthesis. This method enhances enzyme activity and cascade efficiency, leading to a higher yield of this valuable low-calorie sugar.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Metabolic Engineering
Background:
- D-allulose offers significant physiological benefits as a low-calorie rare sugar.
- Current multi-enzyme cascades for d-allulose synthesis suffer from reduced efficiency due to intermediate diffusion and byproduct accumulation.
Purpose of the Study:
- To engineer an efficient artificial in vitro d-allulose biosynthetic pathway using peptide-mediated enzyme complex assembly.
- To mitigate reversible epimerization and enhance substrate channeling in multi-enzyme cascades.
Main Methods:
- Construction of a dual-enzyme complex using d-allulose 6-phosphate epimerase (A6PE) and d-allulose 6-phosphate phosphatase (A6PP) via the ReverseTag/ReverseCatcher system.
- Confirmation of complex assembly using dynamic light scattering and transmission electron microscopy.
- Assembly of a five-enzyme complex (RFE) for an artificial d-allulose biosynthetic pathway.
Main Results:
- The ReverseTag/ReverseCatcher system significantly increased the activity of A6PE (2.1-fold) and A6PP (27.5%).
- The five-enzyme RFE system achieved a d-allulose yield of 63.1% from maltodextrin, a 37.2% improvement over free enzymes.
- Successful spatial organization and improved cascade efficiency were demonstrated.
Conclusions:
- Peptide-mediated covalent assembly is an effective strategy for enhancing multi-enzyme cascade efficiency.
- This approach provides a modular platform for engineering artificial in vitro biosynthetic systems for rare sugar production.
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