Glycosidase-Engineered Microbes for Oligosaccharide Purification

Solution Overview

Problem

Current methods for producing oligosaccharides, particularly Human Milk Oligosaccharides (HMOs), face challenges such as inefficient production, high costs, and the presence of unwanted by-products and intermediates, making large-scale commercial production difficult.

Innovation Solution

The use of glycosidases to degrade unwanted side-products and intermediates during oligosaccharide production, allowing for the purification of desired oligosaccharides in microbial fermentation processes and synthesis reactions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If chemical synthesis or polysaccharide degradation methods are used to produce oligosaccharides, then oligosaccharides can be generated, but unwanted by-products and intermediates are formed requiring complex purification

Engineering Contradiction:

Improveoligosaccharide production efficiencyVSAvoidpurification process complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent applies preliminary action by genetically engineering the producing microorganism to simultaneously express both the oligosaccharide synthesis pathway and the degradation pathways for unwanted by-products and intermediates. This allows the purification function to be built into the production system before the actual oligosaccharide accumulation phase, eliminating the need for separate complex purification steps.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The producing microorganism is engineered to self-purify the oligosaccharide production process by endogenously expressing glycosidases that degrade unwanted by-products and intermediates. The system serves itself by incorporating the purification function within the producing organism, converting a multi-step external purification process into an integrated self-regulating biological system.

Inventive Principle:

Principle #25Self-service

2Manufacturing precision

If multiple purification steps are implemented to remove unwanted oligosaccharides and intermediates, then pure oligosaccharides are obtained, but production costs increase

Engineering Contradiction:

Improveoligosaccharide purityVSAvoidproduction cost

Core Design Contradiction:

Manufacturing precisionVSEase of manufacture

Solution Approach 1:

The producing microorganism is engineered to self-purify the oligosaccharide production process by endogenously expressing glycosidases that degrade unwanted by-products and intermediates. The system serves itself by incorporating the purification function within the producing organism, converting a multi-step external purification process into an integrated self-regulating biological system.

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The patent merges the purification function with the production system by co-expressing synthesis and degradation pathways in the same microorganism. This consolidation integrates multiple previously separate operations (oligosaccharide production and by-product removal) into a single unified biological system, reducing overall process complexity and cost.

Inventive Principle:

Principle #5Merging (Combining)

3Device complexity

If glycosidases are used to degrade unwanted side-products, then purification is simplified, but additional enzyme production is required

Engineering Contradiction:

Improvepurification process simplicityVSAvoidenzyme production requirement

Core Design Contradiction:

Device complexityVSProductivity

Solution Approach 1:

The producing microorganism is engineered to self-purify the oligosaccharide production process by endogenously expressing glycosidases that degrade unwanted by-products and intermediates. The system serves itself by incorporating the purification function within the producing organism, converting a multi-step external purification process into an integrated self-regulating biological system.

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The patent merges the purification function with the production system by co-expressing synthesis and degradation pathways in the same microorganism. This consolidation integrates multiple previously separate operations (oligosaccharide production and by-product removal) into a single unified biological system, reducing overall process complexity and cost.

Inventive Principle:

Principle #5Merging (Combining)

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 enables the efficient production of pure oligosaccharides by removing non-desired compounds, thereby simplifying the purification process and reducing production costs, making large-scale commercial production more viable.

Implementation Method 1

The use of glycosidases to degrade unwanted side-products and intermediates during oligosaccharide production

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Data Source

PatentUS12123040B2Production of oligosaccharides

Publication Date: 2024.10.22 CHR HANSEN AS

AI summary

The present invention relates to the use of one or more glycosidases in the process for the production and/or purification of a produced desired oligosaccharide. The process is preferably a microbial fermentation process using a host microorganism, which may also comprise nucleic acids expressing sugar catabolic pathway proteins suitable for the degradation of saccharides otherwise hindering the purification of the desired oligosaccharide.