Chlorite Dismutase Engineered Cells for Fermentation Sterility
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Solution Overview
Problem
Biotechnological processes face significant challenges in maintaining hygiene due to biological contamination, particularly in large-scale industrial fermentation processes, where contaminating microorganisms such as bacteriophages can disrupt productivity.
Innovation Solution
Recombinantly engineered cells expressing a chlorite dismutase polypeptide are cultured in a medium treated with chlorite, allowing the chlorite dismutase to convert harmful chlorite into harmless chloride and oxygen, thereby reducing or eliminating contaminating microorganisms without harming the host cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chlorite is added to the culture medium to kill contaminating microorganisms, then the hygiene and productivity are improved, but the contaminating microorganisms and their spores are difficult to eliminate completely
Solution Approach 1:
The culture medium is pre-treated with chlorite before inoculation to create a sterile environment, eliminating contaminants in advance. This preliminary disinfection step ensures that when the cultured cells are introduced, the environment is already sanitized, preventing contamination from the outset.
Solution Approach 2:
Chlorite serves as an intermediary substance that mediates between the need for sterilization and the safety of the cultured cells. It acts as a temporary protective agent during the early stages of culture, creating a safe environment that protects vulnerable cultured cells from contaminants while being non-toxic to them.
2Reliability
If conventional disinfectants are used to treat the culture medium, then contaminating microorganisms are reduced, but spores and resistant forms can survive and cause later contamination
Solution Approach 1:
The patent employs multiple disinfection parameters including different chemical agents (chlorite, bleach, alcohol), varying concentrations, exposure times, and temperatures. By changing these parameters in combination, the method overcomes the resistance of spores and resistant forms that would withstand any single disinfection condition.
Solution Approach 2:
The disinfection process uses composite approaches combining multiple chemical disinfectants (chlorite with bleach or alcohol) and physical methods (heat treatment, filtration). This composite strategy creates synergistic effects that eliminate even the most resistant spore forms more effectively than any single method alone.
3Reliability
If the culture medium is treated with strong disinfectants to ensure sterility, then hygiene is improved, but the cultured cells may be damaged or killed by the disinfectant
Solution Approach 1:
Chlorite acts as an intermediary disinfectant that provides effective sterilization while being non-toxic to the cultured cells. It mediates between the need for strong disinfection and cell safety, creating a sterile environment without harming the vulnerable cultured organisms.
Solution Approach 2:
The patent uses controlled concentrations of chlorite and limits exposure time to achieve sterilization at parameters that are sub-lethal to cultured cells. By carefully adjusting these parameters, effective disinfection is achieved while maintaining cell viability.
4Productivity
If the cultured cells are introduced immediately after medium preparation, then productivity is improved, but contamination risk increases due to lack of sterilization time
Solution Approach 1:
The culture medium undergoes preliminary sterilization treatment with chlorite before inoculation, eliminating the need for extended sterilization times after cell introduction. This preliminary action ensures sterility is achieved in advance, allowing immediate inoculation without contamination risk.
Solution Approach 2:
Chlorite serves as a time-saving intermediary that provides rapid sterilization, eliminating the waiting period between medium preparation and inoculation. This mediator enables immediate productivity while maintaining sterility through its fast-acting disinfection properties.
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 method effectively improves fermentation hygiene by reducing or killing contaminating microorganisms while allowing the recombinantly engineered cells to thrive, enhancing the production of fuels and chemicals like hydrocarbons, ethanol, and butanol.
Implementation Method 1
chlorite dismutase, a metabolic enzyme endogenously utilized by perchlorate reducing bacteria, can protect a host cell from the toxic effects of a chlorite-containing disinfectant
Implementation Method 2
the chlorite dismutase to convert harmful chlorite into harmless chloride and oxygen
Implementation Method 3
treating the culture medium with chlorite in an amount sufficient to reduce the growth rate or kill the one or more contaminating microorganisms
Data Source
AI summary
The present disclosure relates to recombinantly engineered cells that contain a chlorite dismutase polypeptide and methods for culturing such cells in a culture medium containing chlorite in an amount sufficient to reduce the growth rate or kill contaminating microorganisms without killing the recombinantly engineered cells. Also provided are methods for the production of a fermentation product using the recombinantly engineered cells.