Human Milk Oligosaccharide Diversification With Transfucosidase Enzymes
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods struggle to produce a diverse range of human milk oligosaccharides (HMOs) efficiently and cost-effectively, particularly in large-scale industrial settings, as they often require costly enzymes and optimal conditions to avoid hydrolysis, limiting the production of complex mixtures resembling the natural variety found in human milk.
Innovation Solution
A method involving the use of enzymes with transfucosidase activity, such as α-trans-fucosidases from specific bacteria, to synthesize fucosylated HMOs, combined with hydrogenolysis to remove protecting groups, allowing for the production of diverse HMOs like 2'-fucosyllactose and 3-fucosyllactose, and their derivatives, in a stereoselective and cost-effective manner.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If existing methods are used to produce HMOs, then production can be achieved, but the cost is high and the diversity of HMOs produced is limited
Solution Approach 1:
The patent segments the HMO production process into modular enzymatic steps using different glycosidases (β-galactosidase, β-N-acetylhexosaminidase, α-L-fucosidase) that can be applied sequentially or in combination. Each enzyme performs a specific transformation on the lactose core or intermediate structures, allowing flexible production of diverse HMOs through combinatorial use of these modular enzymatic tools rather than requiring complete de novo synthesis for each HMO variant.
Solution Approach 2:
The patent employs parameter changes by adjusting enzymatic reaction conditions (pH, temperature, substrate concentration, enzyme-to-substrate ratio) to optimize the production of different HMO types. By varying these parameters, the same enzymatic system can be tuned to produce different proportions of various HMOs, increasing diversity without requiring entirely different production systems for each variant.
2Productivity
If existing methods are used to produce HMOs, then production can be achieved, but the efficiency and scalability to industrial levels are limited
Solution Approach 1:
The patent utilizes the self-service principle by employing enzymatic reactions that automatically catalyze the formation of HMOs from simple precursors like lactose and fucose without requiring complex external intervention. The enzymes naturally drive the reactions toward HMO formation under appropriate conditions, and the system self-regulates through the inherent kinetics and equilibrium of the enzymatic transformations, facilitating efficient and scalable production.
3Reliability
If existing methods are used to produce HMOs, then production can be achieved, but optimal conditions are required to avoid hydrolysis which limits large-scale production
Solution Approach 1:
The patent replaces mechanical/chemical synthesis methods with enzymatic catalysis. Enzymes provide highly specific catalysis that operates under mild conditions (physiological pH and temperature ranges), avoiding the harsh conditions required by traditional chemical synthesis that can cause hydrolysis. This substitution of biochemical catalysis for chemical catalysis enables reliable large-scale production by maintaining product stability while achieving high productivity.
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 and cost-effective production of a variety of fucosylated HMOs, including 2',3-difucosyllactose and fucosylated LNT, on an industrial scale, mimicking the natural complexity of human milk oligosaccharides.
Implementation Method 1
A method involving the use of enzymes with transfucosidase activity, such as α-trans-fucosidases from specific bacteria, to synthesize fucosylated HMOs
Implementation Method 2
combined with hydrogenolysis to remove protecting groups
Data Source
Figure 1
Figure 2
AI summary
A method of diversification of human milk oligosaccharides (HMOs) or precursors thereof, compounds obtainable by the method, and uses and compositions involving such compounds. The method comprises the steps of a) providing at least one compound or a mixture of the compounds selected from the group consisting of: - optionally sialylated and/or fucosylated lactose derivatives of general formula 2 and salts thereof: wherein R is a group removable by hydrogenolysis, R1 independently of each other is fucosyl or H R4 independently of each other is sialyl or H, provided that the compound of general formula 2 is not R-glycoside of lactose, if provided alone; - optionally sialylated and/or fucosylated lactose derivatives of general formula 4 and salts thereof: wherein R1 independently of each other is fucosyl or H R4 independently of each other is sialyl or H, provided that the compound of general formula 4 is not lactose, if provided alone; - lacto-N-tetraose (LNT): - lacto-N-tetraose (LNT) derivatives of the following formula: wherein R is a group removable by hydrogenolysis; - lacto-N-neotetraose (LNnT): - lacto-N-neotetraose (LNnT) derivatives of the following formula: wherein R is a group removable by hydrogenolysis; b) adding at least one enzyme comprising a transglycosidase activity to the at least one compound or a mixture of compounds provided according to step a); and c) incubating the mixture obtained according to step b).