Cell-Free Protein Synthesis Using Pichia pastoris.

Alex J Spice, Rochelle Aw, Karen M Polizzi

Methods in molecular biology (Clifton, N.J.) 2022

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Pichia pastoris (syn. Komagataella phaffii) is an industrially relevant recombinant protein platform that has been used to produce over 5000 proteins to date. Cell-free protein synthesis can be used as a screening tool before strain development or for the production of proteins that are difficult or toxic to make in vivo. Here we describe the methods for generating an active cell lysate from P. pastoris using high pressure homogenization and an improved reaction mix which results in high yields of reporter proteins such as luciferase, and complex proteins such as human serum albumin and virus-like particles. © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Alex J Spice, Rochelle Aw, Karen M Polizzi. Cell-Free Protein Synthesis Using Pichia pastoris. Methods in molecular biology (Clifton, N.J.). 2022;2433:75-88