Ribulose-Phosphate 3-Epimerase Motif Removal to Reduce Psicose Side Reactivity
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Solution Overview
Problem
Existing psicose production methods face inefficiencies and high costs due to the need for additional purification steps to separate fructose from allulose, and known psicose-6-phosphate 3-epimerases exhibit non-specific psicose 3-epimerization activity, making them unsuitable for practical applications.
Innovation Solution
Development of ribulose-phosphate 3-epimerase with specific motifs (motifs I and III) that minimize psicose 3-epimerization activity, allowing for high-yield psicose-6-phosphate production through enzymatic conversion of fructose-6-phosphate, combined with other enzymes for efficient psicose production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If psicose-6-phosphate 3-epimerase is used for psicose production, then psicose-6-phosphate can be produced, but side reactions occur that convert psicose back to fructose, reducing production efficiency
Solution Approach 1:
The patent removes motif II from the enzyme structure to eliminate the harmful side reaction capability. By extracting this specific motif responsible for psicose 3-epimerization, the enzyme retains its primary function of producing psicose-6-phosphate while losing the ability to catalyze the reverse reaction, thus resolving the contradiction between productivity and side reactivity.
Solution Approach 2:
The patent modifies a specific local region (motif II) of the enzyme without affecting the overall structure. By changing the properties of this localized motif through deletion or modification, the enzyme achieves high psicose-6-phosphate production efficiency while minimizing side reactions, demonstrating local quality modification to resolve the contradiction.
2Manufacturing precision
If additional purification steps are added to separate fructose from allulose, then high-purity psicose can be obtained, but production cost and process complexity increase
Solution Approach 1:
The patent performs preliminary action by modifying the enzyme before the reaction to prevent side reactions from occurring in the first place. By deleting motif II from the psicose-6-phosphate 3-epimerase, the enzyme is pre-configured to avoid producing fructose as a side product, thereby eliminating the need for subsequent purification steps and reducing process complexity while maintaining high purity.
3Productivity
If known psicose-6-phosphate 3-epimerase is used, then enzymatic reaction can proceed, but non-specific psicose 3-epimerization activity occurs making it unsuitable for practical production
Solution Approach 1:
The patent extracts motif II from the enzyme sequence, which is responsible for the non-specific psicose 3-epimerization activity. This removal eliminates the harmful side reaction capability while preserving the primary enzymatic function, thereby improving both reliability through enhanced specificity and productivity by preventing reverse reactions.
Solution Approach 2:
The patent changes the structural parameters of the enzyme by deleting or modifying motif II. This parameter change in the enzyme's amino acid sequence fundamentally alters its catalytic properties, making it highly specific for psicose-6-phosphate production while eliminating non-specific activities, thus resolving the contradiction between productivity and reliability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The ribulose-phosphate 3-epimerase achieves high psicose yield with reduced side reactions, enabling cost-effective and efficient psicose production by minimizing psicose 3-epimerization and requiring fewer purification steps.
Implementation Method 1
ribulose-phosphate 3-epimerase achieving high psicose yield with reduced side reactions, enabling cost-effective and efficient psicose production by minimizing psicose 3-epimerization
Data Source
AI summary
Provided are ribulose-phosphate 3-epimerase, a microorganism and a composition, each including the ribulose-phosphate 3-epimerase, and a method of producing psicose-6-phosphate or psicose using the same.