A strain of the Komagataella kurtzmanii yeast, a producer of recombinant peptidase SerP38 from the yellow mealworm Tenebrio molitor, has been obtained. The level of the pro-enzyme secretion was 20-50 mg/L. It was shown that, during secretion in yeast, the target His6-tagged protein was produced in two forms. One of them was a monomer that was efficiently purified by Ni-NTA chromatography and then activated with trypsin. Another form accumulated in the culture medium as oligomers prone to aggregation in the presence of Ni2+ ions and was not activated by trypsin treatment. Aggregation is likely the result of incorrect folding of the polypeptide chain.
Tenebrio molitor, S1 family serine peptidase, SerP38, yeast, Komagataella kurtzmanii, ion-dependent aggregation
Bacterium Hafnia alvei secretes l-methioninase enzyme: Optimization of the enzyme secretion conditions
