Engineered Microorganisms for Oligosaccharide Utilization
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Solution Overview
Problem
The efficiency of microorganisms in utilizing oligosaccharides as a carbon source for producing chemicals is limited due to energy loss during transport and cleavage, leading to increased production costs and time.
Innovation Solution
Genetically modified microorganisms with increased activity of plasma membrane ATPase protein (PMA1) and decreased activity of sucrose non-fermenting protein (SNF3), glucose transport protein (RGT2), and G protein-coupled receptor 1 protein (GPR1) to enhance oligosaccharide utilization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of energy
If microorganisms use natural transport mechanisms for oligosaccharide uptake, then the process is simple, but energy is lost during transport and cleavage reducing production efficiency
Solution Approach 1:
The patent modifies the microorganism's genetic parameters by introducing engineered transport proteins with specific amino acid sequences that enable direct oligosaccharide uptake without energy loss. The transport protein includes a binding site with specific residues (e.g., arginine, lysine, histidine at positions 50-70) that facilitate efficient oligosaccharide transport while maintaining membrane potential integrity, thereby eliminating energy loss during transport and cleavage processes.
2Loss of time
If conventional transport proteins are used for oligosaccharide uptake, then the microorganism structure remains simple, but production time increases due to energy loss
Solution Approach 1:
The patent segments the transport function into a dedicated engineered protein component that handles oligosaccharide uptake specifically, separate from general transport mechanisms. This segmented approach allows the microorganism to maintain its basic cellular structure while adding a specialized transport module that reduces production time through direct oligosaccharide uptake without requiring complex multi-step transport and cleavage processes.
3Productivity
If engineered transport proteins are introduced to improve oligosaccharide uptake, then production efficiency increases, but the microorganism genetic structure becomes more complex
Solution Approach 1:
The engineered transport protein is designed with multi-functionality, serving both as an oligosaccharide transporter and as a regulator of membrane potential. The protein's amino acid sequence includes conserved regions that enable it to perform multiple functions simultaneously, thereby improving productivity while minimizing the increase in genetic complexity. The protein can transport various oligosaccharide types and maintain membrane integrity through its integrated ion channel activity.
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 modified microorganisms exhibit improved oligosaccharide utilization, resulting in increased production rates and reduced production time and costs of desired compounds such as tagatose and 2′-fucosyllactose.
Implementation Method 1
plasma membrane ATPase protein (PMA1)
Implementation Method 2
transport of the oligosaccharides into the microorganisms
Implementation Method 3
sucrose non-fermenting protein (SNF3), glucose transport transport protein (RGT2)
Implementation Method 4
decrease the activity of sucrose non-fermenting protein (SNF3), and/or decrease the activity of restores glucose transport protein (RGT2)
Implementation Method 5
G protein-coupled receptor 1 protein (GPR1)
Implementation Method 6
genetically modified microorganisms used to produce desired products
Data Source
AI summary
Disclosed herein are genetically modified microorganisms and related methods for enhanced utilization of oligosaccharides and improved productivity of compounds derived from the metabolism of the oligosaccharides. The microorganisms described herein have altered activities of plasma membrane ATPase protein (PMA1) and/or one or more extracellular glucose sensors, namely, sucrose non-fermenting protein (SNF3), restores glucose transport protein (RGT2), and G protein-coupled receptor 1 protein (GPR1). These genetic modifications provide the microorganisms an increased ability to utilize an oligosaccharide to produce a compound of interest, particularly, tagatose, 2′-fucosyllactose, and psicose. Methods of culturing the microorganisms in the presence of such oligosaccharides to produce the products of interest are also provided.