High-level expression of hydrolytic enzymes from extremophiles in production hosts such as Escherichia coli and Trichoderma reesei

Hydrolytic enzymes from extreme environments have potential to act as biocatalysts in many industrial applications, such as Biofuels, Oil and Gas, and use of enzymes in household products including detergents.  However, the transfer of genes coding for enzymes from extremophiles have to first undergo certain modifications to suit the expression machinery of the targeted production host.  Modifications such as codon usage change and amino acid substitutions for problematic sites have become routine procedures (1).   

Gene expression relies on strong promoters, design of the expression vector with built-in features to allow targeted secretion and proper folding of the gene product depending on the type of production host.  In the industrial workhorse T. reesei, the strong cbh1 gene promoter is used to drive expression of foreign genes as fusions to the CBH1 protein signal sequence for secretion or to part of the CBH1 mature protein to assist in protein folding as it enters the secretory pathway.

We have developed a pipeline for the high-level expression of different gene products first in E. coli before transferring and expression under multiple promoters in T. reesei (2), for secretion into the cultivation medium, during scale up fermentations.  

(1) Te’o et al., (2000) FEMS Microbiol. Letters. 190: 13-19

(2) Miyauchi et al., (2014) New Biotechnol. 31: 98-103