Malonate Fermentation with Non-Calcium Base pH Control
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Solution Overview
Problem
Existing fermentation processes struggle to efficiently produce malonate in high yields using genetically engineered organisms, particularly due to limitations in metabolic pathways and optimal conditions for carbon source utilization.
Innovation Solution
A fermentation method involving a microorganism with engineered malonate-semialdehyde dehydrogenase and specific carbon sources, along with controlled calcium and pH conditions, to produce malonate at rates of 0.3-5 g L−1 h−1 and yields of at least 40 g/L, utilizing a fermentation medium with calcium ions and a base to optimize malonate production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional fermentation processes are used to produce malonate, then the process is simple, but the yield and production rate are low
Solution Approach 1:
The patent applies parameter changes by optimizing calcium ion concentration (0.05-1.6 g/L), pH level (3.0-6.0), and carbon source composition to achieve high malonate production rates (0.3-5 g L−1 h−1) while maintaining process feasibility
Solution Approach 2:
The patent uses an intermediary approach by introducing a base (other than calcium-containing base) to control pH and facilitate malonate production, acting as a mediator between the carbon source and the desired product
2Productivity
If genetically engineered organisms are used to improve malonate yield, then production efficiency increases, but by-products like ethanol and glycerol increase
Solution Approach 1:
The patent changes physiological parameters by controlling pH (3.0-6.0) and calcium ion concentration (0.05-1.6 g/L) to shift metabolic flux toward malonate production and suppress by-product formation in engineered microorganisms
Solution Approach 2:
The patent implements feedback control by monitoring and adjusting pH and calcium ion levels during fermentation to maintain optimal conditions for malonate production while minimizing by-product formation
3Ease of operation
If calcium-containing bases are used to control pH, then pH control is effective, but malonate production is inhibited
Solution Approach 1:
The patent extracts the harmful component (calcium from calcium-containing bases) while retaining the useful function (pH control) by using alternative bases that do not contain calcium, thereby eliminating the inhibition of malonate production
Solution Approach 2:
The patent uses an intermediary base (non-calcium containing) to mediate pH control without the harmful calcium component, enabling effective pH control while maintaining high malonate production rates
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 method achieves high yields and production rates of malonate, with optional recovery methods enhancing the overall yield to at least 70-99% of the fermentation broth, minimizing by-products like ethanol and glycerol.
Implementation Method 1
Fermentation processes are used commercially at large scale to produce organic molecules such as ethanol, citric acid and lactic acid
Implementation Method 2
The engineered microorganism includes a heterologous malonate-semialdehyde dehydrogenase
Implementation Method 3
calcium ions ranging from 0.05 g/L to 1.6 g/L; and a base other than a calcium containing base
Data Source
AI summary
Various embodiments disclosed relate to a fermentation method for producing malonate. The fermentation method includes a) culturing a microorganism in the presence of a fermentation medium. The fermentation medium includes at least one carbon source; calcium ions ranging from 0.05 g/L to 1.6 g/L; and a base other than a calcium containing base. The fermentation method further comprises b) producing at least 40 grams/liter of malonate. Optionally, the malonate is produced at a rate of at least 0.3 g L−1 h−1 to 5 g L−1 h−1.