Modified Propeptide Regions for Bacillus Protease Secretion
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Solution Overview
Problem
Current methods for producing proteases in bacterial host cells, such as Bacillus species, face inefficiencies in extracellular protein secretion, which hinders the industrial production of enzymes like proteases for use in cleaning, animal feed, and textile processing industries.
Innovation Solution
The use of polynucleotides encoding serine protease sequences with modified or heterologous propeptide regions, combined with expression cassettes and vectors, to enhance the production of mature proteases by modifying the propeptide regions of Bacillus proteases, thereby improving secretion efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used for producing proteases in bacterial host cells, then the production process is simple, but the extracellular protein secretion efficiency is low
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the propeptide region to improve secretion efficiency. Specifically, it replaces amino acids at positions 6, 30, and 32 with alternative residues to optimize extracellular protein secretion while maintaining protease activity. This direct sequence modification resolves the contradiction by enhancing productivity through targeted parameter optimization rather than complex procedural changes.
Solution Approach 2:
The patent applies local quality by focusing modifications specifically on the propeptide region rather than the entire protease structure. By targeting only the propeptide amino acid sequence (particularly positions 6, 30, and 32) while leaving the mature protease region unchanged, the invention improves secretion efficiency locally without requiring complex overall structural changes to the production system.
2Productivity
If the propeptide region is modified to enhance secretion, then the production of mature proteases increases, but the structural complexity of the protease sequence increases
Solution Approach 1:
The patent modifies specific amino acid parameters at key positions (6, 30, and 32) in the propeptide region to enhance mature protease production. By changing only these discrete parameters rather than the entire sequence, the invention increases productivity while minimizing the addition of structural complexity, as the modifications are localized to specific residue positions rather than expanding the overall sequence length or complexity.
Data Source
AI summary
The present invention provides methods and compositions for the production of mature proteases in bacterial host cells. The compositions include polynucleotides encoding serine protease sequences with modified or heterologous propeptide regions; polypeptides comprising serine proteases with modified or heterologous propeptide regions; expression cassettes, DNA constructs, vectors, and chromosomes comprising such polynucleotides; and bacterial host cells comprising such polynucleotides. The methods include methods for enhancing the production of mature proteases in bacterial host cells (e.g. Bacillus sp. host cells). The produced proteases find use in the industrial production of enzymes, suitable for use in various industries, including but not limited to the cleaning, animal feed and textile processing industry.