Bandyopadhyay, S; Morris, SA; Nesbeth, DN; Bracewell, DG; (2021) Lipid reduction to improve clarification and filterability during primary recovery of intracellular products in yeast lysates using exogenous lipase. Journal of Chemical Technology & Biotechnology , 96 (11) pp. 3166-3174. 10.1002/jctb.6871.

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Abstract

BACKGROUND: The yeast Pichia pastoris is a popular host organism for production of a range of biological products, several of which are intracellular. The disruption of yeast cells by homogenisation also releases large quantities of lipids, which can foul the downstream membranes and chromatography matrices used for purification. This work examines lipid removal from yeast cells following homogenisation by enzymatic degradation and its impact on the performance of the subsequent centrifugation and filtration. RESULTS: Lipase treatment of cell homogenate at 37°C for 2 hours, followed by clarification using a scale-down mimic of disc stack centrifugation, resulted in a 6.5-fold improvement in solids removal when compared to untreated feed material. The lipase treated and untreated materials that had undergone initial centrifugation were then tested for filtration performance by passing the material through a 0.45 μm polyethylene sulfone membrane under constant flux. A 50% increase in throughput was observed in comparison to the untreated material. CONCLUSION: This proof-of-concept data suggests enzymatic digestion of lipids, analogous to the widely performed DNA reduction using nucleases, could be a valuable process improvement strategy.

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