Microorganism Trehalose Uptake for Fermentation Yield
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Solution Overview
Problem
In industrial fermentations of Corynebacterium glutamicum, significant amounts of trehalose are excreted and not reused, leading to losses in product formation and biomass concentration, as well as contamination in the fermentation supernatant.
Innovation Solution
Increasing the expression of a polynucleotide coding for a protein subunit of the ABC transporter with trehalose importer activity, allowing the microorganism to actively uptake trehalose from the medium, thereby reducing its accumulation and utilizing it for metabolic purposes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional fermentation processes are used, then L-amino acid production is achieved, but significant amounts of trehalose are excreted and not reused, leading to loss of substrate carbon and reduced product yield
Solution Approach 1:
The patent converts the harmful effect of trehalose accumulation (which represents wasted substrate carbon and reduced productivity) into a beneficial effect by engineering the microorganism to uptake and metabolize the accumulated trehalose. The overexpression of the msiK gene encodes a sugar import protein that actively transports trehalose back into the cell, where it can be metabolized through the trehalose metabolism pathway to produce additional biomass and L-amino acids, thus converting the waste product into a useful resource.
Solution Approach 2:
The patent applies parameter changes by modifying the expression level of the msiK gene through overexpression. This changes the transport capacity parameter of the microorganism, enabling it to actively uptake trehalose at rates sufficient to prevent external accumulation. The altered expression parameter transforms the microorganism's metabolic behavior from excreting trehalose to actively consuming it, thereby improving substrate utilization efficiency and product yield.
2Quantity of substance
If conventional fermentation processes are used, then L-amino acid production is achieved, but biomass concentration is reduced due to unused substrate carbon
Solution Approach 1:
The patent converts the energy loss from unused substrate carbon (trehalose) into beneficial biomass production. By enabling trehalose uptake through msiK overexpression, the microorganism recovers the carbon and energy that would otherwise be wasted, channeling it through central metabolism to generate additional ATP and precursor molecules for biomass synthesis, thereby increasing overall biomass concentration.
Solution Approach 2:
The engineered microorganism performs self-service by autonomously taking up and metabolizing its own waste product (trehalose). The overexpressed MsiK transporter automatically imports trehalose from the external medium into the cell without requiring external intervention, and the cell's endogenous trehalose metabolism pathway processes it to generate energy and building blocks for biomass production, making the system self-sufficient in eliminating its metabolic byproduct.
3Reliability
If conventional fermentation processes are used, then L-amino acid production is achieved, but contamination in the product supernatant occurs
Solution Approach 1:
The patent converts the contamination problem caused by trehalose accumulation in the supernatant into a benefit by eliminating the accumulation itself. Through msiK overexpression, trehalose is continuously imported and metabolized, preventing its buildup in the external medium. This eliminates the contamination issue without requiring additional purification steps, as the harmful substance is converted into useful metabolic intermediates and energy within the cell.
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 enhances substrate carbon utilization, increases biomass and product yield in amino acid production, and minimizes contamination in the fermentation broth.
Implementation Method 1
Increasing the expression of a polynucleotide coding for a protein subunit of the ABC transporter with trehalose importer activity, allowing the microorganism to actively uptake trehalose from the medium
Implementation Method 2
In industrial fermentations of Corynebacterium glutamicum, significant amounts of excreted trehalose are found in the supernatant. The externally accumulated trehalose is not returned to the metabolic cycle by the cells.
Data Source
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AI summary
The invention relates to a microorganism which produces and/or secretes an organo-chemical compound, the microorganism demonstrating an expression of one or more protein subunits of the ABC transporter with the activity of a trehalose importer, which expression is increased in comparison with the respective starting strain, where the microorganism is capable of taking up trehalose from the medium; and to a process for the production of an organo-chemical compound using the microorganism according to the invention, where an accumulation of trehalose in the fermentation broth is reduced or avoided.