Cysteine-Rich Insecticidal Peptide Production for Proper Folding
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Solution Overview
Problem
Existing insecticidal peptides face challenges in commercialization due to issues such as toxicity, production difficulties, cost, degradation, improper folding in genetically modified organisms, and resistance from insect strains, particularly Bacillus thuringiensis (Bt) resistant insects, limiting their use in crop protection.
Innovation Solution
Development of special expression cassettes and systems for proper folding of insecticidal peptides in plants, combination of Cysteine Rich Insecticidal Peptides (CRIPS) and Pore Forming Insecticidal Proteins (PFIPS) to enhance toxicity and activity, and methods to produce these peptides at higher yields and lower costs, including use of Endoplasmic Reticulum Signal Peptides (ERSP) and Translational Stabilizing Proteins (STA) for targeted expression and synergistic effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If insecticidal peptides are produced in genetically modified organisms, then production capability is improved, but proper folding and stability deteriorate
Solution Approach 1:
The patent introduces a signal peptide as an intermediary element that mediates between the production system and the folding process. The signal peptide directs the nascent peptide chain to the endoplasmic reticulum where proper folding occurs, resolving the contradiction between high production capability and proper folding stability in genetically modified organisms.
Solution Approach 2:
The patent replaces complex mechanical folding control systems with a biochemical signaling system. Instead of attempting to mechanically control folding conditions in GM organisms, the invention uses signal peptides that automatically direct peptides to appropriate cellular compartments (endoplasmic reticulum) where folding occurs naturally, simplifying the production system while ensuring proper folding.
2Reliability
If conventional insecticides are used, then immediate pest control is improved, but insect resistance develops
Solution Approach 1:
The patent changes the fundamental parameter of insecticide structure by using synthetic peptides with specific amino acid sequences rather than conventional chemical insecticides. This structural parameter change creates a new mode of action that bypasses existing insect resistance mechanisms, maintaining pest control effectiveness while reducing adaptability of resistant strains.
Solution Approach 2:
The patent creates composite peptide structures combining multiple amino acid residues with specific properties (hydrophobic, hydrophilic, charged residues) to form a complex three-dimensional structure. This composite structure at the molecular level provides multiple interaction points with the target site, making it harder for insects to develop resistance compared to single-molecule conventional insecticides.
3Productivity
If insecticidal peptides are produced at high yields, then production cost is reduced, but production complexity increases
Solution Approach 1:
The patent segments the production process into distinct functional modules: signal peptide for targeting, mature peptide for activity, and specific amino acid sequences for folding. This segmentation allows each component to be independently optimized and produced at high yields through standardized molecular biology techniques, reducing overall production complexity despite high productivity requirements.
Data Source
AI summary
New insecticidal proteins, nucleotides, peptides, their expression in plants, methods of producing the peptides, new processes, production techniques, new peptides, new formulations, and new organisms, a process which increases the insecticidal peptide production yield from yeast expression systems. The present invention is also related and discloses selected endotoxins we call cysteine rich insecticidal peptides (CRIPS) which are peptides derived from Bacillus thuringiensis (Bt) and their genes and endotoxins in combination with toxic peptides known as Inhibitor Cystine Knot (ICK) genes and peptides as well as with other types of insecticidal peptides such as trypsin modulating oostatic factor (TMOF) peptide sequences used in various formulations and combinations; of both genes and peptides, useful for the control of insects.