Human Type 17 Collagen Polypeptides via E. Coli Expression
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Solution Overview
Problem
Current methods for producing human collagen face challenges such as high production costs, long cycles, and risks of viral infection and sensitization from animal-derived collagen, while conventional methods result in collagen with lost biological activity and thrombosis risks, limiting its application in biomedicine and tissue engineering.
Innovation Solution
Recombinant expression of human type 17 collagen polypeptides (C17A3, C17B3, C17C1) using an optimized E. coli expression system, allowing for low-cost, large-scale production of water-soluble polypeptides with high stability and cell adhesion activity, suitable for medical devices, tissue engineering, and cosmetics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If animal-derived collagen is used, then collagen products can be obtained, but there is risk of viral infection and sensitization
Solution Approach 1:
The patent uses recombinant DNA technology to copy the human collagen gene sequence and express it in E. coli bacteria, creating a synthetic version of human collagen that mimics the natural protein's structure and function without using animal tissues. This eliminates the risk of viral infection and sensitization associated with animal-derived collagen while maintaining the desired collagen properties.
2Quantity of substance
If conventional extraction methods are used, then collagen can be obtained from animal tissues, but the collagen loses its original biological activity
Solution Approach 1:
The patent replaces conventional mechanical and chemical extraction methods with biological recombinant expression. Instead of using acid, alkali, and enzymatic hydrolysis that destroy biological activity, the invention uses genetically engineered E. coli to produce collagen through natural biological synthesis, preserving the protein's native structure and biological functionality.
3Reliability
If conventional recombinant expression methods are used, then human-derived collagen can be expressed, but production cost is too high and production cycle is too long
Solution Approach 1:
The patent optimizes multiple parameters of the recombinant expression system: selecting a simplified collagen sequence (C17A3/C17B3/C17C1) for faster expression, using E. coli as the host organism for rapid reproduction and low-cost cultivation, adjusting cultivation conditions for high-yield production, and streamlining the purification process. These parameter changes collectively reduce production time and cost while maintaining human collagen authenticity.
4Ease of manufacture
If collagen with strong coagulation function is used, then collagen can be prepared by conventional methods, but there is great risk of thrombosis in tissue engineering products
Solution Approach 1:
The patent extracts and expresses only the essential collagen domain (C17A3/C17B3/C17C1) from the full collagen protein, separating the cell adhesion and structural functions from the coagulation function. This minimalistic approach retains the beneficial properties needed for tissue engineering while eliminating or reducing the thrombosis risk associated with full-length collagen's strong coagulation activity.
Data Source
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AI summary
Provided is a polypeptide, a production method therefor and a use thereof. The polypeptide includes 63 to 1496 continuous amino acid residues of SEQ ID NO: 9, and includes a sequence represented by (A) m or is composed of the sequence represented by (A)m, wherein each A is an amino acid sequence selected from any one of those represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, or an amino acid sequence of any one of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 with 1 or more, for example 2, 3, 4 or 5, amino acid residues substituted, added, or deleted, or a sequence having 83%-97% sequence identity with an amino acid sequence represented by any one of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; m is an integer between 1 and 10; and each A is the same or different and two adjacent As are directly connected by peptide bonds or connected by at least one amino acid residue. The polypeptide has a cell adhesion activity.