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Microorganisms 2018, 6(1), 3; https://doi.org/10.3390/microorganisms6010003

Pichia pastoris is a Suitable Host for the Heterologous Expression of Predicted Class I and Class II Hydrophobins for Discovery, Study, and Application in Biotechnology

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON MSE 3E5, Canada
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Received: 13 November 2017 / Revised: 15 December 2017 / Accepted: 29 December 2017 / Published: 5 January 2018
(This article belongs to the Special Issue Filamentous Fungi in White Biotechnology)
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Abstract

The heterologous expression of proteins is often a crucial first step in not only investigating their function, but also in their industrial application. The functional assembly and aggregation of hydrophobins offers intriguing biotechnological applications from surface modification to drug delivery, yet make developing systems for their heterologous expression challenging. In this article, we describe the development of Pichia pastoris KM71H strains capable of solubly producing the full set of predicted Cordyceps militaris hydrophobins CMil1 (Class IA), CMil2 (Class II), and CMil3 (IM) at mg/L yields with the use of 6His-tags not only for purification but for their detection. This result further demonstrates the feasibility of using P. pastoris as a host organism for the production of hydrophobins from all Ascomycota Class I subdivisions (a classification our previous work defined) as well as Class II. We highlight the specific challenges related to the production of hydrophobins, notably the challenge in detecting the protein that will be described, in particular during the screening of transformants. Together with the literature, our results continue to show that P. pastoris is a suitable host for the soluble heterologous expression of hydrophobins with a wide range of properties. View Full-Text
Keywords: hydrophobin; Pichia pastoris; immunoassay; protein expression; fermentation; Cordyceps militaris hydrophobin; Pichia pastoris; immunoassay; protein expression; fermentation; Cordyceps militaris
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Gandier, J.-A.; Master, E.R. Pichia pastoris is a Suitable Host for the Heterologous Expression of Predicted Class I and Class II Hydrophobins for Discovery, Study, and Application in Biotechnology. Microorganisms 2018, 6, 3.

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