Engineered Microorganisms Utilizing Melamine Nitrogen
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
In the fermentation industry, maintaining a robust monoculture environment is challenging due to opportunistic infections and metabolic burdens, as existing methods like antibiotic addition are often undesirable and ineffective in preventing contaminant growth.
Innovation Solution
Genetically engineering organisms with non-native enzymes such as allophanate hydrolase, biuret amidohydrolase, and melamine deaminase to convert nitrogen-containing compounds like triazine and melamine into products, providing a competitive advantage and reducing contaminant growth.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If antibiotic addition is used to prevent contaminant growth, then the fermentation environment becomes more robust, but spontaneous resistant contaminations occur and antibiotic use is often undesirable
Solution Approach 1:
The invention converts the harmful presence of contaminants into a beneficial selective pressure by engineering the host organism to exclusively utilize a specific nitrogen source (e.g., melamine, triazine) that contaminants cannot metabolize. This creates a growth advantage for the engineered host without requiring antibiotics, thereby preventing contaminant growth while avoiding antibiotic resistance issues.
Solution Approach 2:
The invention changes the nitrogen source parameter in the fermentation medium from conventional sources (ammonium salts, urea) to unconventional nitrogen-containing compounds (melamine, triazine, cyanuric acid). This parameter change creates a selective environment where only the engineered host with specific metabolic pathways can grow, eliminating the need for antibiotics and preventing resistant contaminations.
2Productivity
If conventional nitrogen sources are used in cell culture media, then all nutrients necessary for host cell growth are provided, but potential contaminating organisms can also grow in this environment
Solution Approach 1:
The invention applies local quality by providing a specialized nitrogen source that only the engineered host can utilize. The host organism is equipped with specific enzymes (e.g., melamine deaminase, triazine phosphoribonucleotidohydrolase) that enable it to metabolize the unconventional nitrogen source, while contaminants lacking these enzymes cannot grow. This creates a localized growth advantage for the host in the fermentation environment.
Solution Approach 2:
The invention converts the potential harm of contaminant growth into a benefit by using the nitrogen-containing compound as a selective agent. The compound itself is not harmful to the host (which can metabolize it), but it selectively inhibits contaminant growth, thereby protecting the fermentation process without requiring additional antibiotics or growth inhibitors.
3Reliability
If genetically engineered organisms are used to utilize unconventional nitrogen sources, then competitive advantage against contaminants is achieved, but the organism must be successfully transformed with specific nucleic acid sequences
Solution Approach 1:
The invention achieves universality by using a plasmid vector system that can deliver multiple functional genes (melamine deaminase, triazine phosphoribonucleotidohydrolase, and selectable marker genes) in a single transformation event. This multi-functional plasmid approach simplifies the genetic engineering process compared to introducing multiple separate genes, while providing the host with all necessary capabilities to utilize unconventional nitrogen sources and resist contaminants.
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 creates a robust fermentation environment by enabling the engineered organisms to utilize challenging nitrogen sources, minimizing contaminant growth and eliminating the need for prophylactic antibiotics, thus enhancing production efficiency and product yield.
Implementation Method 1
the non-native gene encodes for a non-native enzyme selected from the group consisting of allophanate hydrolase, biuret amidohydrolase, cyanuric acid amidohydrolase, guanine deaminase, melamine deaminase, isopropylammelide isopropylaminohydrolase, cyanamide hydratase, urease, and urea carboxylase
Data Source
AI summary
Disclosed are genetically engineered organisms, such as yeast and bacteria, that have the ability to metabolize atypical nitrogen sources, such as melamine and cyanamide. Fermentation methods using the genetically engineered organisms are also described. The methods of the invention are robust processes for the industrial bioproduction of a variety of compounds, including commodities, fine chemicals, and pharmaceuticals.