Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation

• DOI: 10.18517/ijaseit.12.3.15109
• Publication Date: 2022-06-29T09:58:46Z
• Authors: Eko Wahyu Putro; Dini Nurdiani; - Hariyatun; Nuruliawaty Utami; Wien Kusharyoto; Ario Betha Juanss...; Rozy Ayu Silvia
• Keywords: Fed-batch culture

Production of sufficient insulin at a more affordable price is necessary. The increase in the number people living with diabetes puts burden on healthcare and economy. P. pastoris promising host to produce human precursors high yield minimal medium secretes low levels endogenous protein impurities. precursor involves several parameters, including glycerol concentration, culture density, methanol composition. This study evaluated effect those parameters expression flask culture. Subsequently, fermentation bioreactor was carried according information obtained from Methanol feeding induce undertaken by pulses fed-batch modes. method modified standard technique incorporating constant flow rates variable feed concentrations. Cell density determined based optical measurement 600 nm dry cell weight. Tricine SDS-PAGE reversed-phase HPLC conducted analysis. pulse produced higher concentrations ~445 mg/L than ~267 mg/L. However, can be an alternative producing when 100% (v/v) difficult apply.