High cell density fed-batch production of insecticidal recombinant ribotoxin hirsutellin A from Pichia pastoris.

ABSTRACT:

Background

The fungal ribotoxin hirsutellin A (HtA) exhibits strong insecticidal activity; however, efficient systems for expressing recombinant HtA (rHtA) are lacking. Here, we established an efficient heterologous expression system to produce large amounts of rHtA.

Results

Recombinant Pichia pastoris transformants with high levels of secretory rHtA were screened, and in a fed-batch reactor, rHtA was secreted at levels up to 80 mg/l following methanol induction, which was more than sixfold higher than that in shake flasks. Approximately 7 mg of highly pure rHtA was obtained from 300 ml of fed-batch culture supernatant by Ni⁺-nitriloacetic acid affinity chromatography and CM Sepharose ion-exchange chromatography. Mass spectrometry results revealed rHtA as a native

SUBMITTER: Li H 

PROVIDER: S-EPMC6195745 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature