OTA Detoxifying Polypeptides with Enhanced Thermostability
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Solution Overview
Problem
Current methods for detoxifying ochratoxin A (OTA) in food and feed materials are inadequate due to insufficient activity, stability, and recombinant reducibility of existing OTA-hydrolyzing enzymes, making them unsuitable for real-life applications in the food and feed production line.
Innovation Solution
Development of polypeptides with at least 70% sequence identity to specific amino acid sequences, possessing peptidase activity EC 3.4.13.9, which are capable of effectively hydrolyzing OTA, along with their variants that have alterations such as substitutions, deletions, or insertions, ensuring improved thermostability and OTA degradation efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing OTA-hydrolyzing enzymes are used for detoxification, then OTA degradation is achieved, but the enzymes exhibit insufficient activity, stability, and recombinant reducibility
Solution Approach 1:
The patent applies parameter changes by modifying amino acid residues at specific positions (e.g., positions 167, 168, 218, 275) in the OTA-hydrolyzing enzyme sequence. These sequence modifications result in variant enzymes with improved thermostability and catalytic activity, resolving the contradiction between enzyme stability and degradation efficiency.
Solution Approach 2:
The patent creates variant copies of the original OTA-hydrolyzing enzyme through site-directed mutagenesis. By copying the enzyme structure with specific amino acid substitutions, the patent generates improved variants that maintain OTA hydrolysis capability while enhancing stability and activity for industrial application.
2Temperature
If existing OTA-hydrolyzing enzymes are used, then OTA detoxification is possible, but the enzymes show insufficient thermostability for industrial applications
Solution Approach 1:
The patent modifies enzyme parameters through amino acid substitutions at critical positions (e.g., I167G, I168G, F218Y, Y275V) to enhance thermostability. These parameter changes allow the enzyme to maintain structural integrity and catalytic activity at elevated temperatures required for industrial food and feed processing.
3Ease of manufacture
If existing OTA-hydrolyzing enzymes are applied, then some OTA degradation occurs, but the enzymes lack sufficient recombinant reducibility for manufacturing
Solution Approach 1:
The patent creates recombinant copies of the modified enzyme sequences with optimized codon usage and structural features. This enables efficient expression in heterologous host systems while maintaining the enhanced thermostability and catalytic activity, facilitating scalable manufacturing for industrial detoxification applications.
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 proposed polypeptides and their variants demonstrate enhanced thermostability and OTA degradation ability, making them suitable for industrial applications in detoxifying OTA in food and feed materials, thereby ensuring safer food and feed supplies.
Implementation Method 1
The enzyme termed ochratoxinase from Aspergillus niger was previously reported to possess OTA hydrolyzing activity
Implementation Method 2
said polypeptide/s having peptidase activity having EC 3.4.13.X
Data Source
AI summary
The present invention relates to novel uses of the polypeptides and novel variants of said polypeptides capable of detoxifying Ochratoxin A (OTA) and methods (e.g., for detoxifying mycotoxins) based thereon. The present invention further relates to compositions, kits, transgenic plants, transgenic seeds, transgenic pollen grains, foodstuff, intermediate foodstuff; fodder, intermediate fodder; feed, intermediate feed; additive (e.g., foodstuff-, fodder- or feed additive), intermediate additive (e.g., foodstuff-, fodder- or feed intermediate additive); detoxifying agent, intermediate detoxifying agent; nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic and/or mixture/s thereof comprising one or more of the polypeptides and/or variants capable of detoxifying Ochratoxin A (OTA).