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毕赤酵母中重组蛋白生产的菌株和分子工具。

Strains and Molecular Tools for Recombinant Protein Production in Pichia pastoris.

机构信息

Austrian Centre of Industrial Biotechnology (ACIB), Graz, Austria.

Bisy GmbH, Hofstaetten/Raab, Austria.

出版信息

Methods Mol Biol. 2022;2513:79-112. doi: 10.1007/978-1-0716-2399-2_6.

Abstract

Within the last two decades, the methylotrophic yeast Pichia pastoris (Komagataella phaffii) has become an important alternative to E. coli or mammalian cell lines for the production of recombinant proteins. Easy handling, strong promoters, and high cell density cultivations as well as the capability of posttranslational modifications are some of the major benefits of this yeast. The high secretion capacity and low level of endogenously secreted proteins further promoted the rapid development of a versatile Pichia pastoris toolbox. This chapter reviews common and new "Pichia tools" and their specific features. Special focus is given to expression strains, such as different methanol utilization, protease-deficient or glycoengineered strains, combined with application highlights. Different promoters and signal sequences are also discussed.

摘要

在过去的二十年中,甲醇营养型酵母毕赤酵母(Komagataella phaffii)已成为生产重组蛋白的大肠杆菌或哺乳动物细胞系的重要替代品。这种酵母的主要优点包括易于操作、强启动子、高密度培养以及能够进行翻译后修饰。该酵母还具有高分泌能力和低水平的内源性分泌蛋白,这进一步促进了多功能毕赤酵母工具盒的快速发展。本章综述了常见和新型“毕赤工具”及其特点。特别关注表达菌株,如不同甲醇利用、蛋白酶缺陷或糖基工程菌株,并结合应用亮点进行讨论。不同的启动子和信号序列也进行了讨论。

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