Amide Compound Biosynthesis With 6-Aminocaproate Transporters
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Solution Overview
Problem
Microbial production of nylon intermediates like 6-aminocaproic acid, caprolactam, and hexamethylenediamine is hindered by undesired enzymatic activities that produce byproducts and impurities, increasing costs and reducing efficiency.
Innovation Solution
Engineering non-naturally occurring microbial organisms with specific genetic modifications, including exogenous transporters and disruptions of endogenous transporters, to enhance the production of these intermediates by reducing carbon-competing intermediates and byproducts.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If engineered microorganisms are used to produce nylon intermediates, then production of desired products is achieved, but undesirable byproducts are produced due to undesired enzymatic activity
Solution Approach 1:
The patent removes undesired enzymatic activities from the microbial cell by deleting specific genes (e.g., gabD, ybfF, yghD, yjgB, yahK, gabT) that encode enzymes producing harmful byproducts. This extraction of harmful functions allows the cell to focus on producing desired nylon intermediates without contamination from byproducts.
Solution Approach 2:
The patent modifies the metabolic parameters of the microbial cell by altering gene expression levels and metabolic pathway fluxes. Through genetic engineering, the cell's metabolic capacity is redirected toward desired products while suppressing pathways that generate byproducts, optimizing the ratio of desired to harmful products.
2Productivity
If engineered microorganisms produce nylon intermediates, then yield is improved, but cost and complexity of biosynthesis increase
Solution Approach 1:
The patent simplifies the biosynthesis process by removing complex regulatory networks and unnecessary enzymatic steps that lead to byproduct formation. By deleting genes responsible for harmful pathways, the overall metabolic network is streamlined, reducing the complexity of the biosynthesis process while maintaining high yield.
Solution Approach 2:
The patent discards harmful byproducts and unnecessary metabolic pathways, focusing resources only on the essential steps for producing desired nylon intermediates. This selective approach eliminates wasted metabolic capacity and simplifies the overall biosynthesis process.
3Productivity
If engineered microorganisms produce nylon intermediates, then production efficiency is improved, but byproducts and impurities increase
Solution Approach 1:
The patent extracts and removes the enzymatic activities that generate byproducts and impurities. By deleting specific genes (gabD, ybfF, yghD, yjgB, yahK, gabT), the cell loses the capacity to produce harmful substances, thereby improving the purity of nylon intermediates while maintaining biosynthesis efficiency.
Solution Approach 2:
The patent changes the metabolic parameters of the cell to optimize both efficiency and purity. By altering gene expression and metabolic flux distribution through genetic engineering, the cell achieves high production efficiency while simultaneously improving product purity by redirecting metabolic pathways away from byproduct formation.
Data Source
AI summary
Disclosed are biosynthetic methods and engineered microorganisms that enhance or improve the biosynthesis of 6-aminocaproate, hexamethylenediamine, caproic acid, caprolactone, or caprolactam. The engineered microorganisms are modified to include, for example, upredulated and/or exogenous transporters for 6-aminocaproate, deletions and/or downregulated importers for 6-aminocaproate, upregulated and/or exogenous glutamate dehydrogenase, and/or deletions and/or downregulation of rcsA and/or cpsBG. Other engineered microorganisms may have disruptions of endogenous transporters for 6-aminocaproate.