Modified Protease Pro Region Mutations for Bacillus Secretion
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Solution Overview
Problem
Current methods for extracellular protein secretion in microorganisms, such as Bacillus species, are inefficient and require optimization to enhance the production of industrially valuable proteases like serine proteases.
Innovation Solution
Modified polynucleotides encoding modified proteases with mutated pro regions are introduced into microorganisms, specifically Bacillus species, to enhance protease production by altering the secretion and maturation processes, resulting in increased enzymatic activity and production levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional secretion methods are used in microorganisms, then protein secretion occurs, but the efficiency and production levels are insufficient
Solution Approach 1:
The invention applies parameter changes by modifying the amino acid sequence of the pro region in the protease precursor. Specific substitutions at positions equivalent to 33, 37, 57, and 61 in the V049 protease pro region alter the secretion and maturation parameters, resulting in enhanced protease production levels and secretion efficiency compared to the wild-type enzyme.
2Productivity
If the pro region is left unmodified, then the protease follows natural maturation pathway, but production levels remain limited
Solution Approach 1:
The invention applies local quality by making targeted modifications only to the pro region of the protease precursor while leaving the mature protease sequence unchanged. This localized approach modifies specific amino acid positions (equivalent to 33, 37, 57, and 61) to enhance production without requiring complete redesign of the entire polynucleotide sequence.
Data Source
Figure 1A~1B
Figure 1C~1D
Figure 2A~2D
AI summary
This invention relates to modified polynucleotides encoding modified proteases, and methods for altering the production of proteases in microorganisms. In particular, the present invention relates to methods for altering the expression of proteases in microorganisms, such as Bacillus species. The invention discloses modified polynucleotides, vectors, modified polypeptides, and processes for enhancing the production of proteases.