Talaromyces Cellulolyticus Pep4 Attenuation for Higher Protein Secretion
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Solution Overview
Problem
Existing methods for producing proteins using Talaromyces cellulolyticus do not consider the relationship between the Pep4 protein and protein production, limiting the efficiency of protein production.
Innovation Solution
Modify Talaromyces cellulolyticus to reduce the activity of the Pep4 protein by reducing or disrupting the pep4 gene expression, optionally combined with reducing the activities of YscB and CreA proteins, to enhance protein-producing ability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the activity of endogenous proteases is reduced to improve protein production, then the productivity of heterologous proteins is improved, but the microorganism's ability to degrade and process proteins is reduced
Solution Approach 1:
The patent converts the harmful effect of endogenous proteases (which degrade produced proteins) into a beneficial strategy by selectively reducing their activity. By attenuating specific protease genes (pep4, yscB, creA) while maintaining other cellular functions, the microorganism's protein degradation capability is reduced, thereby improving the stability and accumulation of produced proteins without completely eliminating proteolytic functions
Solution Approach 2:
The patent applies parameter changes by modifying the activity levels of specific protease enzymes through genetic manipulation. By creating strains with reduced expression or disruption of pep4, yscB, and creA genes, the protease activity parameters are adjusted to optimal levels that maximize protein production while maintaining cellular viability and other essential functions
2Productivity
If multiple gene modifications are combined to reduce protease activities, then the protein production ability is significantly enhanced, but the genetic engineering complexity increases
Solution Approach 1:
The patent merges multiple gene modification strategies into a single integrated approach. By simultaneously targeting pep4, yscB, and creA genes in combination, the patent creates a synergistic effect where the combined reduction of multiple protease activities produces a greater improvement in protein production than individual modifications alone, while establishing a standardized multi-gene modification protocol
Data Source
AI summary
A method for producing a protein is provided. An objective protein is produced by culturing in a culture medium Talaromyces cellulolyticus having an objective protein-producing ability, which has been modified so that the activity of a Pep4 protein is reduced.