Ire1 Variant Engineering for Higher Fungal Protein Secretion
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need for alternatives to enhance the secretion of heterologous proteins in filamentous fungal host cells to improve productivity and yield.
Innovation Solution
A variant Ire1 polypeptide with an amino acid substitution at position 153 of the Trichoderma reesei Ire1 polypeptide is introduced, which improves the secretion of heterologous polypeptides by mutating the ire1 gene to encode a variant Ire1 polypeptide with at least 70% to less than 100% sequence identity to the native sequence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If the native Ire1 polypeptide is used in filamentous fungal host cells, then the host cell maintains normal cellular function, but the productivity and yield of secreted heterologous polypeptides are limited
Solution Approach 1:
The invention applies parameter changes by mutating specific amino acid residues in the Ire1 polypeptide sequence. The patent identifies and modifies key residues (such as K153E, K153D, K153R, K153Q, K153N, K153H, K153A, K153S, K153T, K153C mutations) to alter the functional properties of Ire1, thereby enhancing its ability to promote secretion of heterologous proteins while maintaining cellular viability.
Solution Approach 2:
The invention applies local quality by making specific localized mutations at particular amino acid positions (especially position 153 and other identified residues) within the Ire1 polypeptide. Rather than globally altering the entire protein, the patent focuses modifications on specific critical regions that govern secretion activity, thereby improving heterologous protein secretion while preserving essential cellular functions.
2Productivity
If the native Ire1 polypeptide is used in filamentous fungal host cells, then the host cell maintains normal cellular function, but the yield of secreted heterologous polypeptides is limited
Solution Approach 1:
The invention applies parameter changes by mutating specific amino acid residues in the Ire1 polypeptide sequence. The patent identifies and modifies key residues (such as K153E, K153D, K153R, K153Q, K153N, K153H, K153A, K153S, K153T, K153C mutations) to alter the functional properties of Ire1, thereby enhancing its ability to promote secretion of heterologous proteins while maintaining cellular viability.
Solution Approach 2:
The invention applies local quality by making specific localized mutations at particular amino acid positions (especially position 153 and other identified residues) within the Ire1 polypeptide. Rather than globally altering the entire protein, the patent focuses modifications on specific critical regions that govern secretion activity, thereby improving heterologous protein secretion while preserving essential cellular functions.
Data Source
AI summary
The present invention relates to an isolated variant Ire1 polypeptide, comprising (a) an amino acid substitution at position 153 of the Trichoderma reesei Ire1 polypeptide of SEQ ID NO: 2; or (b) an amino acid substitution at a position corresponding to position 153 of the Trichoderma reesei Ire1 of SEQ ID NO: 2 and has at least 70%, but less than 100%, sequence identity to the amino acid sequence of SEQ ID NO: 2. The present invention also relates to a recombinant filamentous fungal host cell comprising the mutated ire1 gene encoding the variant Ire1 polypeptide for producing a secreted heterologous polypeptide of interest, a method for producing the secreted heterologous polypeptide of interest in the recombinant filamentous fungal host cell, and a method of improving the productivity or yield of the secreted heterologous polypeptide of interest in the recombinant filamentous fungal host cell.