Genetic Constructs for the Production of Heterologous Proteins of Interest

Genetic constructs for the rapid and efficient expression and high‑level production of proteins of interest in filamentous fungi, reducing costs and meeting the needs of industrial protein manufacturing.

Market need: Despite significant advances in recombinant DNA technology over recent decades, which allow efficient protein production, industrial‑scale manufacturing still faces important limitations. These include the difficulty of achieving simultaneous and stable expression of multiple proteins within a single cell, the inherent unpredictability of genetic manipulations, and the high costs associated with using inducers to activate gene expression. In addition, these inducers may be unstable, interfere with cellular metabolism, and negatively impact both process performance and product quality. Therefore, there is a clear need for expression systems that do not rely on such inducers and that enable more stable, cost‑effective, and efficient protein production.

Proposed solution: Description of genetic constructs comprising a promoter sequence from a gene encoding a P‑type sodium and/or potassium pump, whose expression level increases as the extracellular pH rises, operably linked to a nucleotide sequence encoding a protein of interest.
By increasing the pH of the culture medium of cells into which these constructs have been introduced, the expression and production of high levels of proteins of interest are induced, without the need to use inducers that cause significant alterations to cellular metabolism.

Competitive advantages

Fast method compatible with high ammonium levels.
Allows the use of complex media based on low‑cost carbon and nitrogen sources.
Enables the co‑expression of two different proteins in the same host cell.
A more alkaline pH in continuous culture prevents pellet formation, reducing the energy required to agitate the medium in the fermenter.