Fusarium Toxin-Cleaving Polypeptide Variants for Stable Detoxification
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Solution Overview
Problem
Current methods for detoxifying Fusarium toxins, such as those used in food and feed processing, require oxygen and cofactors, and existing enzymes lack sufficient temperature stability, making them ineffective at elevated temperatures during processes like pelleting and bioethanol production.
Innovation Solution
Development of Fusarium toxin-cleaving polypeptide variants with enhanced temperature stability and activity, specifically modified carboxylesterase enzymes that can cleave Fusarium toxins into non-toxic products without oxygen or cofactors, allowing for effective detoxification at elevated temperatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing carboxylesterase enzymes are used for detoxifying Fusarium toxins, then the detoxification reaction can proceed without oxygen or cofactors, but the enzymes lack sufficient temperature stability and become ineffective at elevated temperatures during processing
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid positions in the carboxylesterase enzyme sequence. Specific positions (66, 199, 302, 364, 377, 389, 394, 424, 430, 463, 465, 469) were identified and mutated to improve temperature stability while preserving detoxification activity. This rational design approach changed the enzyme's physical parameters to achieve stability at elevated temperatures without requiring oxygen or cofactors.
2Ease of manufacture
If traditional detoxification methods requiring oxygen and cofactors are used, then the enzymatic reaction can proceed under mild conditions, but the process becomes complex and less suitable for industrial processing at elevated temperatures
Solution Approach 1:
The patent extracts and eliminates the dependency on oxygen and cofactors from the detoxification process by using a modified carboxylesterase enzyme that catalyzes the hydrolysis of Fusarium toxins independently. This removes the complex requirements for oxygen supply and cofactor addition, simplifying the manufacturing process while enabling operation at elevated temperatures during industrial processing.
3Reliability
If the enzyme sequence is modified to improve temperature stability, then the enzyme can withstand elevated temperatures, but the modification process increases complexity in identifying and testing specific amino acid positions
Solution Approach 1:
The patent applies local quality by focusing modifications on specific, predetermined amino acid positions within the enzyme sequence rather than random mutagenesis. The identified positions (66, 199, 302, 364, 377, 389, 394, 424, 430, 463, 465, 469) represent localized regions that, when modified, confer temperature stability. This targeted approach reduces the overall complexity by concentrating efforts on key positions rather than exhaustive screening.
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 modified polypeptide variants demonstrate increased temperature stability and activity, enabling safe and reliable detoxification of Fusarium toxins during processes like pelleting and bioethanol production, ensuring food and feed safety without the need for oxygen or cofactors.
Implementation Method 1
The carboxylesterases used and the polypeptide variants thereof hydrolytically split off at least one tricarballylic acid from the Fusarium toxin, whereby the toxic effects of the Fusarium toxin are removed or reduced
Implementation Method 2
The invention relates to the use of carboxylesterases and polypeptide variants thereof for the cleavage of at least one fusarium toxin
Data Source
AI summary
The invention relates to fusarium toxin-cleaving polypeptide variants of a fusarium toxin carboxyl esterase with the SEQ ID NO:46. Each of the polypeptide variants has an amino acid sequence shortened by 47 amino acids at the N terminus, and the amino acid sequences have at least 70%, preferably 80%, in particular 100%, sequence identity, namely SEQ ID NO:1, to the amino acid sequence portion 48 - 540 of the SEQ ID NO:46. The invention also relates to isolated polynucleotides which code for the polypeptide variants, to a fusarium toxin-cleaving additive containing at least one polypeptide variant and optionally at least one auxiliary agent, to the use of the polypeptide variants or the additive, and to a method for hydrolytically cleaving at least one fusarium toxin.