Engineering a novel secretion signal for cross-host recombinant protein expression

Area of Science:

• Molecular Biology
• Biotechnology
• Cell Biology

Background:

• Protein secretion is crucial for many cellular functions and biotechnological applications.
• Conventional secretion signals are typically hydrophobic and located at the N-terminus.
• Efficient and versatile secretion systems are needed for recombinant protein production.

Purpose of the Study:

• To identify and characterize a novel secretion signal (SS) for directing recombinant protein secretion.
• To demonstrate the broad applicability of the SS in diverse prokaryotic and eukaryotic hosts.
• To evaluate the inducibility and expression dynamics of the SS.

Main Methods:

• Construction and testing of fusion reporter proteins with the novel SS.
• Expression and secretion analysis in Escherichia coli, Saccharomyces cerevisiae, and six eukaryotic cell lines.
• Estrogen-inducible expression and secretion studies in mammalian cells.

Main Results:

• The novel SS successfully directed secretion of fusion reporter proteins in E. coli, yeast, and six eukaryotic cell lines.
• Estrogen-inducibility of secretion was demonstrated in multiple eukaryotic cell lines.
• Secretion rate was rapid, correlating with intracellular mRNA levels.
• Secretion in bacteria and yeast was dependent on culture conditions and induction rate.

Conclusions:

• The novel SS provides a flexible and efficient strategy for recombinant protein production and secretion.
• This system facilitates convenient and rapid studies of protein expression across various hosts.
• The SS has significant potential for biotechnological applications requiring controlled protein secretion.