The development of a heterologous gene expression system in thermophilic fungus Thermoascus aurantiacus

Thermoascus aurantiacus is a thermophilic fungus that belongs to the ascomycetous class and has attracted increasing interest for its ability to produce thermostable cellulolytic enzymes and growth at elevated temperatures. However, studies on this organism have been limited because of the lack of a genetic manipulation system. Here, we developed a polyethylene glycol (PEG)-mediated transformation system for T. aurantiacus based on an orotidine-5'-monophosphate decarboxylase (pyrG)-deficient mutant, with this method achieving a transformation efficiency of 33 ± 3 transformants per microgram of DNA. Intracellular or secretory expression of heterologous proteins, including green fluorescent protein, β-galactosidase and α-amylase, in T. aurantiacus was successful under the inducible endogenous cellobiohydrolase and endoglucanase gene promoter or the constitutive heterologous pyruvate decarboxylase and enolase gene promoter from Trichoderma reesei. To the best of our knowledge, this is the first report on PEG-mediated transformation of T. aurantiacus, which sets the foundation for strain improvement for biotechnological applications and functional genomic studies.

The online version contains supplementary material available at 10.1007/s13205-021-02963-w.