Bacillus Host Cells for Enhanced Protease Production
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Solution Overview
Problem
Current methods for enhancing protease production in Bacillus strains, such as screening for hyper-producing strains and optimizing fermentation technologies, have limitations, and there is a need for additional means to increase protease yield effectively.
Innovation Solution
Genetically modified Bacillus sp. host cells with at least one inactivated phr and/or rap gene, combined with overexpression of YmaH, to enhance the production of proteins of interest, particularly proteases like subtilisin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods (screening hyper-producing strains, optimizing fermentation) are used to enhance protease production, then production levels can be improved, but the improvement is limited and additional means are needed to achieve further increases in protease yield
Solution Approach 1:
The patent removes (inactivates) specific phr and/or rap genes from the Bacillus genome to eliminate their inhibitory effect on protease production. This extraction of harmful genetic elements directly increases protease yield without adding complex genetic constructs, resolving the contradiction between improving productivity and maintaining simplicity.
Solution Approach 2:
The patent changes the genetic parameters of the Bacillus host by inactivating specific genes (phr and/or rap) to alter the regulatory network controlling protease production. This parameter change in the genetic makeup enables higher protease production levels while using a relatively simple gene knockout approach rather than complex fermentation optimization.
2Productivity
If phr and/or rap genes are inactivated to increase protein production, then production levels improve, but the genetic modification process becomes more complex
Solution Approach 1:
The patent applies gene knockout techniques to remove or inactivate specific phr and/or rap genes from the Bacillus genome. This extraction of inhibitory genetic elements simplifies the regulatory network and directly enhances protein production, making the genetic modification straightforward compared to adding complex expression systems.
Solution Approach 2:
Instead of adding genes or regulatory elements to increase protein production, the patent inverts the approach by removing (inactivating) genes that suppress protein production. This inversion strategy simplifies the genetic modification process while achieving the desired increase in productivity.
3Productivity
If YmaH is overexpressed in modified host cells, then protein production is further increased, but the number of genetic modifications increases
Solution Approach 1:
The patent combines two genetic modifications into a unified strategy: inactivating phr and/or rap genes to remove inhibition, and overexpressing YmaH to enhance production. This merging of negative regulation removal with positive regulation addition creates a synergistic effect that further increases protein production while consolidating the genetic modification approach.
Solution Approach 2:
The patent changes multiple genetic parameters simultaneously - inactivating repressor genes (phr/rap) and overexpressing a positive regulator (YmaH). This coordinated parameter change in the genetic network achieves cumulative enhancement of protein production, with each modification building on the previous one to maximize productivity.
Data Source
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AI summary
The present invention provides host cells that have been genetically manipulated to have an enhanced capacity to produce proteins of interest. In particular, the invention relates to modified Bacillus sp. Host cells that have at least one inactivated phr gene. The enhanced production of proteins of interest by the modified Bacillus sp. Host cells is further increased in modified Bacillus sp. Host cells that overexpress YmaH. Methods for producing proteins of interest in the modified host cells are also provided