Cellooligosaccharide Synthesis Using Mixed Solvent Control

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Solution Overview

Problem

Conventional enzymatic synthesis of cellooligosaccharides using cellodextrin phosphorylase in aqueous solutions results in a wide degree-of-polymerization distribution, leading to the production of cellooligosaccharides with higher than average degrees of polymerization, which is undesirable for improved performance.

Innovation Solution

The method involves reacting α-glucose-1-phosphate with glucose, cellobiose, or alkylated glucose using cellodextrin phosphorylase in a mixed solvent containing water and a water-soluble organic solvent, such as methanol or dimethyl sulfoxide, to suppress the formation of high-degree cellooligosaccharides and achieve a narrower polymerization distribution.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If water is used as a reaction solvent in enzymatic synthesis of cellooligosaccharides, then the enzyme maintains its catalytic function, but the degree-of-polymerization distribution becomes wide and high-degree cellooligosaccharides are formed

Engineering Contradiction:
Improvecatalytic functionVSAvoiddegree-of-polymerization distribution
Core Design Contradiction:
ReliabilityVSManufacturing precision

Solution Approach 1:

The patent changes the physical-chemical parameters of the reaction solvent by introducing a water-soluble organic solvent (such as dimethyl sulfoxide, methanol, or ethanol) to create a mixed solvent system. This parameter change modifies the solvent properties to control the polymerization reaction, resulting in a narrow degree-of-polymerization distribution while maintaining enzyme activity. The organic solvent concentration is optimized to achieve the desired balance between enzyme stability and product distribution control.

Inventive Principle:
Principle #35Parameter changes

2Manufacturing precision

If a mixed solvent containing water-soluble organic solvent is used, then the degree-of-polymerization distribution is narrowed and high-degree cellooligosaccharides are suppressed, but the enzyme may be denatured and inactivated

Engineering Contradiction:
Improvedegree-of-polymerization distributionVSAvoidcatalytic function
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

The patent carefully optimizes the concentration parameters of the water-soluble organic solvent in the mixed solvent system. By controlling the organic solvent content within specific ranges (e.g., 1-50% v/v depending on the specific solvent), the patent achieves narrow degree-of-polymerization distribution while maintaining sufficient enzyme activity. This parameter optimization resolves the contradiction between product precision and enzyme stability.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The water-soluble organic solvent acts as an intermediary substance that mediates between the aqueous enzyme environment and the polymerization reaction. It modifies the solvent properties to control polymer chain growth without directly denaturing the enzyme, thereby enabling precise control over the degree of polymerization while preserving catalytic function.

Inventive Principle:
Principle #24Intermediary (Mediator)

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

This approach effectively limits the formation of high-degree cellooligosaccharides, resulting in products with a more controlled degree-of-polymerization distribution and improved selectivity, as demonstrated by MALDI-TOF-MS analysis.

Implementation Method 1

reacting α-glucose-1-phosphate and at least one primer selected from the group consisting of glucose, cellobiose, and alkylated glucose with cellodextrin phosphorylase

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

a synthesis method using the reverse reaction of cellodextrin phosphorylase, a phosphorolytic enzyme

Methodology Applied
Scientific EffectPhosphorolytic reaction: Chemical Bonding

Data Source

PatentEP3789493B1Cellooligosaccharide production method
Publication Date: 2023.02.15 DKS CO LTD
  • EP3789493B1 patent drawingFigure 1~2
  • EP3789493B1 patent drawingFigure 3~4
  • EP3789493B1 patent drawingFigure 5~7

AI summary

A method for producing a cellooligosaccharide that enables the formation of a cellooligosaccharide having a high degree of polymerization to be suppressed in enzymatic synthesis of a cellooligosaccharide, the method comprising reacting α-glucose-1-phosphate and at least one primer selected from the group consisting of glucose, cellobiose, and alkylated glucose with cellodextrin phosphorylase in a mixed solvent containing water and a water-soluble organic solvent.