Effect of two preservation methods on the viability and enzyme production of a recombinant Komagataella phaffii (Pichia pastoris) strain

In this study, we evaluated the effect of glycerol and skim milk in cryopreservation at -80 °C, as well as the use of skim milk or its combination with NaCl, disaccharides or sorbitol in freeze-drying on the cell viability and activity of a recombinant lysosomal enzyme (i.e., human β-hexosaminidase-A) produced in K. phaffii GS115 strain. The results showed that cryopreservation with glycerol and skim milk, as well as freeze-drying using disaccharides and sorbitol with skim milk, maintained the viability above 80%. Although variations in enzyme activity among treatments were found, the use of disaccharides had a positive effect on the enzymatic activity levels. This is the first report of the evaluation of two suitable methods to preserve a recombinant K. phaffii strain, preventing the loss of viability and maintaining the activity of the recombinant protein.PMID:34951975 | DOI:10.1016/j.cryobiol.2021.12.004

Source: Cryobiology - December 24, 2021 Category: Biology Authors: Angela Mar ía Alvarado-Fernández Edwin Alexander Rodr íguez-López Angela Johana Espejo-Mojica Angela Roc ío Mosquera-Arévalo Carlos Javier Alm éciga-Díaz Alba Alicia Trespalacios-Rangel Source Type: research