NSP24 Signal Sequence for Heterologous Protein Secretion
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Solution Overview
Problem
Current methods for expressing and secreting heterologous proteins in host cells like yeast, filamentous fungi, and bacteria are inefficient, particularly in achieving biologically active mature forms, and lack effective signal peptide sequences for protein secretion.
Innovation Solution
The use of a signal sequence from an aspartic protease obtained from Trichoderma, referred to as the NSP24 signal sequence, is employed to facilitate the expression and secretion of heterologous proteins such as phytase, glucoamylase, and cellulase by fusion with these proteins, utilizing expression vectors and host cells like Trichoderma and Aspergillus species.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional signal peptide sequences are used for heterologous protein expression, then protein secretion is achieved, but secretion efficiency and biological activity are low
Solution Approach 1:
The patent changes the key parameter of signal peptide sequence by replacing conventional signal peptides with the NSP24 signal peptide sequence from Trichoderma reesei. This parameter change results in significantly improved secretion efficiency while maintaining biological activity of the heterologous proteins, directly resolving the technical contradiction between productivity and reliability.
2Quantity of substance
If heterologous proteins are expressed intracellularly, then protein production is achieved, but purification complexity increases
Solution Approach 1:
The patent extracts the heterologous protein from intracellular compartments by using the NSP24 signal peptide to direct secretory pathway targeting. This extraction of the protein into the extracellular medium simplifies purification processes while maintaining production quantity, as the protein is naturally secreted into the culture medium rather than remaining trapped inside cells.
Solution Approach 2:
The NSP24 signal peptide acts as an intermediary that mediates the transport of heterologous proteins from the intracellular synthesis site to the extracellular secretion medium. This intermediary function enables efficient protein transfer across the cell membrane, achieving both high production and simplified purification by naturally directing proteins to the extracellular environment.
Data Source
AI summary
The invention relates to heterologous polypeptide expression and secretion by filamentous fungi and vectors and processes for expression and secretion of such polypeptides. More particularly, the invention discloses the use of a signal sequence form an aspartic protease obtained from Trichoderma and referred to as an NSP24 signal sequence.