Mannose-Specific Glycoprotease for Yeast Agglomeration
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Solution Overview
Problem
Current technologies lack the ability to effectively exploit mannose-modified proteins for commercial purposes, particularly in yeast and fungal cells, where this form of O-linked glycosylation is prevalent.
Innovation Solution
Development of recombinant polypeptides with mannose-specific glycoprotease activity that can specifically target and modify mannose-decorated amino acid sequences in proteins, allowing for linker-specific cleavages and agglomeration of yeast and fungal cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If mannose-specific glycoprotease activity is used to target mannose-decorated amino acid sequences, then linker-specific cleavages are achieved, but the complexity of the system increases
Solution Approach 1:
The patent uses mannose-specific glycoprotease as an intermediary enzyme that recognizes and cleaves specifically at mannose-decorated amino acid sequences in linker regions. This intermediary enables precise cleavage without requiring complex engineering of the target proteins themselves, resolving the contradiction between precision and complexity.
Solution Approach 2:
The invention changes the chemical parameter of the linker region by introducing mannose decorations on serine or threonine residues. This parameter change creates a specific recognition target for the glycoprotease, enabling precise cleavage through biochemical specificity rather than mechanical or physical means.
2Productivity
If proteases are used to modify mannose-decorated amino acid sequences, then protein glycosylation is altered, but the loss of carbohydrate content reduces protein stability
Solution Approach 1:
The patent segments the protein structure by specifically cleving only the linker regions containing mannose decorations, while leaving the core functional domains intact. This segmentation approach allows modification of specific regions without compromising the overall stability of the protein structure.
Solution Approach 2:
The glycoprotease acts locally on specific mannose-decorated serine or threonine residues in linker regions, rather than uniformly processing all carbohydrate content. This local quality approach enables selective modification of linker sequences while preserving the stability-critical regions of the protein.
3Quantity of substance
If yeast and fungal cells are agglomerated using mannose-specific glycoprotease, then fermentation by-products are enhanced, but the loss of time for aggregation process increases
Solution Approach 1:
The mannose-specific glycoprotease treatment enables yeast cells to self-aggregate through natural adhesion mechanisms following enzymatic modification of surface mannose decorations. This self-service approach eliminates the need for external mechanical aggregation devices or complex multi-step processes, reducing time loss while achieving protein enrichment.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The use of these recombinant polypeptides enables efficient agglomeration of yeast and fungal cells, resulting in a fermentation product with reduced carbohydrate content and increased protein enrichment, thereby enhancing the value of fermentation by-products.
Implementation Method 1
the modification is proteolysis
Implementation Method 2
recombinant polypeptide having mannose-specific glycoprotease activity
Implementation Method 3
the contacting results in aggregation of the cells, cell bodies or cellular components
Implementation Method 4
method for agglomerating organisms displaying mannose-decorated amino acid sequences on their surface
Data Source
AI summary
Disclosed are compositions and methods involving proteases specific for mannose-modified proteins. The compositions and methods are particularly useful for making linker-specific cleavages in proteins produced by yeast and fungal cells. One use of the compositions and methods is for agglomerating yeast and yeast components in fermentation products. Another use of the composition is for producing a fraction of protein with reduced carbohydrate content.