Lactose Import Engineering for High-Yield HMO Biosynthesis

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Solution Overview

Problem

The limited availability and contamination risks associated with chemical synthesis of human milk oligosaccharides (HMOs) necessitate the development of a cost-effective, large-scale biosynthetic production method that optimizes the transport of lactose precursors and reduces side product formation.

Innovation Solution

A genetically engineered cell overexpresses lactose permease genes in combination with heterologous MFS transporters and glycosyltransferases to enhance the transport and biosynthesis of HMOs, particularly LNT and LNnT, while maintaining a favorable cellular equilibrium to reduce side products.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If chemical synthesis is used to produce HMOs, then production scalability is achieved, but contamination risk and cost increase

Engineering Contradiction:

Improveproduction scalabilityVSAvoidcontamination risk

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent replaces chemical synthesis methods with a biological system (genetically engineered bacteria) to produce HMOs. The bacterial cell uses enzymatic pathways (glycosyltransferases) to synthesize HMOs from lactose precursors, eliminating the need for harmful chemicals and high-pressure reaction conditions associated with chemical synthesis, while maintaining scalability through microbial fermentation

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent modifies cellular parameters by overexpressing specific transport proteins (lactose permease LacY and MFS transporters) and glycosyltransferase enzymes to optimize HMO production. This involves changing the expression levels of genetic components to enhance lactose import and HMO synthesis efficiency, thereby improving productivity while maintaining biosynthetic cleanliness

Inventive Principle:

Principle #35Parameter changes

2Productivity

If lactose import is enhanced through overexpression of lactose permease, then HMO production increases, but cellular equilibrium is disrupted and side products form

Engineering Contradiction:

ImproveHMO productionVSAvoidside product formation

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent combines multiple functional components into a coordinated system: lactose permease (LacY) for lactose import, MFS transporters (YberC, Nec, Vag) for HMO export, and glycosyltransferases for HMO synthesis. This integrated approach ensures that lactose import, HMO production, and HMO secretion work together to maintain cellular equilibrium and prevent side product accumulation

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent employs regulatory mechanisms where the expression of transport proteins and enzymes is controlled by promoters responsive to cellular conditions. The system monitors and adjusts lactose import and HMO synthesis rates to maintain optimal cellular equilibrium, preventing excessive side product formation through feedback control of metabolic pathways

Inventive Principle:

Principle #23Feedback

3Productivity

If multiple sugar transporters are expressed to optimize lactose and HMO transport, then production efficiency increases, but device complexity increases

Engineering Contradiction:

Improveproduction efficiencyVSAvoidtransporter system complexity

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent utilizes the MFS transporter family members (YberC, Nec, Vag) that can transport multiple substrates including HMOs and other sugars. These transporters serve multiple functions: HMO export, lactose transport, and maintenance of cellular osmotic balance, reducing the need for separate dedicated transporters for each function and thereby managing system complexity

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 increases the overall yield and reduces by-product formation, making it suitable for large-scale production with simplified purification processes and lower costs.

Implementation Method 1

overexpression of the E. coli, lactose permease LacY... enhances the transport of lactose and produced HMO(s)

Methodology Applied

Scientific EffectActive transport:

Implementation Method 2

express the glycosyltransferases needed for the synthesis of the desired oligosaccharides

Methodology Applied

Scientific EffectEnzymatic catalysis: Enzyme

Implementation Method 3

Fermentation based processes have been developed for several HMO(s)... utilize genetically engineered bacterial strains

Methodology Applied

Scientific EffectFermentation: Fermentation

Data Source

PatentUS20260015636A1Enhancing formation of human milk oligosaccharides (HMOS) by modifying lactose import in the cell

Publication Date: 2026.01.15 DSM IP ASSETS BV

AI summary

This invention relates to a method of producing one or more human milk oligosaccharides (HMOs), in particular LNT and/or LNnT, in a genetically engineered cell comprising an enhanced oligosaccharide transport capability. The genetically modified cell comprises a series of genetic modification which enable the production of one or more HMO(s), and a series of genetic modification that enhances the transport of lactose and produced HMO(s).