Recombinant Signal Sequence Viscosity Control
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Solution Overview
Problem
The biotechnology industry faces challenges in large-scale, cost-effective manufacture, recovery, and purification of recombinant proteins, particularly in balancing host cell growth and expression, protein location, and selective recovery and purification, with existing methods struggling to optimize these factors effectively.
Innovation Solution
A recombinant signal sequence comprising the sequence MGRISSGG or variants thereof, combined with a heterologous signal sequence, is used to control viscosity and basal expression of recombinant proteins during fermentation, enabling efficient production and recovery by optimizing protein secretion and reducing broth viscosity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Shape
If conventional signal sequences are used to direct protein secretion, then protein location is improved, but broth viscosity increases and basal expression is difficult to control
Solution Approach 1:
The patent modifies the signal sequence parameters by introducing specific amino acid substitutions (particularly at positions 3-5 of the signal sequence) to optimize the balance between secretion efficiency and broth viscosity control. This parameter optimization allows the signal sequence to maintain its protein-targeting function while reducing adverse effects on fermentation broth properties.
Solution Approach 2:
The patent creates a composite signal sequence structure that combines elements known to promote secretion with modifications that reduce viscosity. The hybrid signal sequence integrates functional domains from different sources or designs, achieving synergistic effects that simultaneously improve protein location control and minimize harmful viscosity increases.
2Productivity
If signal sequences promote protein secretion, then productivity is improved, but basal expression becomes difficult to control
Solution Approach 1:
The patent optimizes signal sequence parameters including amino acid composition, hydrophobicity, and length to achieve reliable control of basal expression. Specific modifications at key positions allow the signal sequence to remain dormant under non-inducing conditions while maintaining high secretion efficiency upon induction, thus improving both productivity and reliability.
3Productivity
If recombinant protein expression is increased, then productivity is improved, but fermentation broth viscosity increases
Solution Approach 1:
The patent employs parameter optimization of the signal sequence to decouple protein expression levels from broth viscosity increases. By fine-tuning the signal sequence characteristics, the system achieves high productivity while maintaining fermentation broth properties suitable for downstream processing and scale-up.
Data Source
AI summary
The present invention relates to a recombinant signal sequence derived from E. coli. The invention also relates to a fusion protein comprising the signal sequence, a recombinant protein and methods of producing the recombinant protein. The recombinant signal sequence can be used to provide a method for controlling the viscosity of the fermentation, and/or controlling basal (pre-induction) expression of the recombinant protein.