Engineered Methanotrophic Bacteria Multi-Carbon Substrate Utilization
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Solution Overview
Problem
Methanotrophic bacteria are limited by the low solubility of methane, leading to slow growth and restricted industrial applications, as they rely solely on methane as a carbon and energy source, necessitating the development of strains that can utilize alternative, inexpensive multi-carbon substrates.
Innovation Solution
Genetically engineered methanotrophic bacteria are created to express exogenous nucleic acids encoding multi-carbon substrate utilization pathways, enabling them to grow on substrates like glucose, acetate, lactate, arabinose, citrate, succinate, and glycerol as primary or sole carbon sources, using components such as glucose transporters, acetate transporters, lactate dehydrogenase, and glycerol kinase.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If methanotrophic bacteria rely solely on methane as carbon and energy source, then they maintain their natural metabolic simplicity, but their growth is limited by low methane solubility resulting in slow growth and low cell density
Solution Approach 1:
The patent introduces multiple exogenous nucleic acids encoding different substrate utilization pathways (glycerol, glucose, acetate, lactate, arabinose, citrate, succinate) into methanotrophic bacteria, enabling these organisms to utilize diverse multi-carbon substrates in addition to methane. This multi-functional capability allows the bacteria to overcome methane solubility limitations while maintaining their natural methane metabolism, thereby achieving high productivity through alternative carbon sources.
2Productivity
If methanotrophic bacteria are genetically engineered to utilize multiple substrates, then productivity and biomass production improve, but device complexity and genetic modification requirements increase
Solution Approach 1:
The patent divides the complex substrate utilization capability into separate functional modules, with each exogenous nucleic acid encoding a specific substrate utilization pathway (glycerol kinase for glycerol, glucose transporter for glucose, acetate transporter for acetate, etc.). This segmentation allows for systematic genetic engineering where each pathway can be independently introduced and optimized, reducing the overall complexity compared to attempting to engineer a single universal pathway.
3Ease of operation
If methanotrophic bacteria use methane as sole carbon source, then their metabolic pathway remains simple, but industrial applications are hampered by safety concerns with explosive substrate and slow growth
Solution Approach 1:
The patent fundamentally changes the carbon substrate parameter from methane (C1) to multi-carbon substrates (C2-C6), which have higher solubility and faster metabolic rates. This parameter change transforms the growth kinetics and substrate handling characteristics, enabling faster biomass production and eliminating safety concerns associated with methane's explosive properties while maintaining the bacteria's methanotrophic identity through retained methane utilization capability.
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
These engineered bacteria can achieve high biomass production and rapid growth on diverse carbon sources, overcoming the limitations of methane reliance and expanding their industrial applicability.
Implementation Method 1
at least one exogenous nucleic acid encoding a multi-carbon substrate utilization pathway component, wherein the at least one exogenous nucleic acid is expressed in a sufficient amount to permit growth of the non-naturally occurring methanotrophic bacteria on the multi-carbon substrate as a primary carbon source
Implementation Method 2
components such as glucose transporters, acetate transporters, lactate dehydrogenase, and glycerol kinase
Data Source
AI summary
The present disclosure relates to genetically engineered methanotrophic bacteria with the capability of growing on a multi-carbon substrate (e.g., glucose) as a primary or sole carbon source and methods for growing methanotrophic bacteria on the multi-carbon substrate.