Acetyl-CoA Synthase Variant for High CO Tolerance
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Solution Overview
Problem
Current technologies face challenges in increasing carbon monoxide availability and metabolic engineering for acetogen microorganisms, particularly in high CO concentration environments, due to limited genetic engineering systems and low productivity of 4-carbon organic substances like butyric acid.
Innovation Solution
Development of an acetyl-CoA synthase variant by substituting histidine at position 636 with another amino acid in the Eubacterium limosum microorganism, enhancing CO availability and tolerance, and incorporating mutations in the CODH/ACS complex to improve carbon monoxide fixation and tolerance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If acetogen microorganisms are used to produce 4-carbon organic substances from waste gas, then carbon monoxide conversion is achieved, but productivity is extremely limited due to low growth at high CO concentrations
Solution Approach 1:
The invention changes the amino acid sequence parameter of the ACS protein by substituting histidine at position 636 with lysine, arginine, or ornithine. This parameter change in the protein structure enables the microorganism to maintain self-growth capability at high CO concentrations (60-100%), thereby resolving the contradiction between productivity and reliability by allowing the organism to thrive in high CO environments where it can efficiently produce 4-carbon organic substances
2Productivity
If genetic engineering is performed on acetogen to increase metabolite productivity, then metabolite production is improved, but the complexity of establishing a suitable genetic engineering system increases
Solution Approach 1:
The invention modifies the ACS protein by substituting specific amino acids (histidine at position 636 with lysine, arginine, or ornithine) to create a variant that enables high CO tolerance. This targeted parameter change in the protein sequence provides a straightforward genetic engineering approach that avoids complex system establishment, directly improving metabolite productivity while maintaining relatively simple engineering procedures
3Reliability
If microorganisms are exposed to high CO concentration stress to evolve CO tolerance, then CO availability is improved, but the complexity of the evolution process increases
Solution Approach 1:
The invention directly introduces the beneficial parameter change (amino acid substitution at position 636 of ACS protein) into the microorganism through genetic engineering, achieving CO tolerance without requiring complex multi-generational evolution processes. This approach simplifies the process by directly implementing the key parameter change that confers CO tolerance, avoiding the complexity of maintaining and selecting through extensive evolution experiments
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 variant microorganism exhibits improved growth and productivity under high CO concentrations, enabling the production of industrially valuable compounds and efficient removal of waste gases, with enhanced CO fixation and tolerance capabilities.
Implementation Method 1
acetyl-CoA synthase variant, in which histidine at position 636 from the N-terminus of acetyl-CoA synthase derived from a microorganism of the genus Eubacterium limosum is substituted with another amino acid
Data Source
AI summary
The present invention relates to a microorganism with increased carbon monoxide availability and use thereof.