Bacillus Licheniformis Strain Engineering for Higher Protein Secretion
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Solution Overview
Problem
There is a need for Bacillus licheniformis strains with enhanced protein production capabilities to optimize industrial protein yields, particularly for enzymes like proteases and amylases, while avoiding toxic by-products and ensuring safety compliance.
Innovation Solution
Genetic modification of Bacillus licheniformis cells by introducing a native prsA promoter operably linked to a native prsA open reading frame, optionally with deletions or disruptions of dltA and rghR2 genes, to enhance protein production, and fermenting under suitable conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional B. licheniformis strains are used for protein production, then the production process is simple and safe (QPS/GRAS status), but the protein yield is limited
Solution Approach 1:
The patent applies parameter changes by modifying specific genetic parameters of B. licheniformis strains. It introduces a second copy of the prsA gene under its native promoter to increase protein secretion capacity, and deletes the dltA gene to modify cell wall properties for enhanced protein release. These targeted genetic parameter changes increase protein yield while maintaining the safety profile of the organism.
Solution Approach 2:
The patent segments the protein production process into distinct genetic components: the prsA gene copy for secretion, the dltA deletion for cell wall modification, and the protein of interest expression cassette. This segmentation allows independent optimization of each component and simplifies the overall strain construction process through modular genetic engineering.
2Productivity
If protein production is optimized for high yield, then industrial biotechnology applications are improved, but the complexity of strain modification increases
Solution Approach 1:
The patent employs parameter changes by systematically modifying key genetic parameters: adding a prsA gene copy to enhance secretion capacity, deleting dltA to improve cell wall permeability for protein release, and optimizing the protein expression cassette. These parameter changes are implemented using standard molecular biology techniques, making the strain modification process relatively straightforward despite the productivity gains.
3Productivity
If the prsA gene is introduced to enhance protein secretion, then protein production increases, but the genetic modification complexity increases
Solution Approach 1:
The patent applies the copying principle by introducing a second copy of the native prsA gene into the B. licheniformis genome. This copy is placed under the control of the native prsA promoter, creating an additional source of the secretion protein without requiring complex heterologous expression systems. The copying approach simplifies genetic modification while effectively increasing protein secretion capacity.
Data Source
AI summary
The present disclosure is generally related to compositions and methods for constructing and/or obtaining B. licheniformis cells (e.g., protein production hosts) comprising enhanced protein production capabilities. Thus, certain embodiments are related to genetically modified Bacillus licheniformis strains derived from parental B. licheniformis strains producing increased amounts of one or more proteins of interest.