Advances in Using Hansenula polymorpha as Chassis for Recombinant Protein Production.

João Heitor Colombelli Manfrão-Netto,Nádia Skorupa Parachin,Antônio Milton Vieira Gomes

doi:10.3389/fbioe.2019.00094

João Heitor Colombelli Manfrão-Netto, Nádia Skorupa Parachin + Show 1 more

Open Access

https://doi.org/10.3389/fbioe.2019.00094

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Abstract

The methylotrophic yeast Hansenula polymorpha, known as a non-conventional yeast, is used for the last 30 years for the production of recombinant proteins, including enzymes, vaccines, and biopharmaceuticals. Although a large number of reviews have been published elucidating the applications of this yeast as a cell factory, the latest was released about 10 years ago. Therefore, this review aimed at summarizing available information on the use of H. polymorpha as a host for recombinant protein production in the last decade. Examples of chemicals and virus-like particles produced using this yeast also are discussed. Firstly, the aspects that feature this yeast as a host for recombinant protein production are highlighted including the techniques available for its genetic manipulation as well as strategies for cultivation in bioreactors. Special attention is given to the novel genomic editing tools, mainly CRISPR/Cas9 that was recently established in this yeast. Finally, recent examples of using H. polymorpha as an expression platform are presented and discussed. The production of human Parathyroid Hormone (PTH) and Staphylokinase (SAK) in H. polymorpha are described as case studies for process establishment in this yeast. Altogether, this review is a guideline for this yeast utilization as an expression platform bringing a thorough analysis of the genetic aspects and fermentation protocols used up to date, thus encouraging the production of novel biomolecules in H. polymorpha.

Highlights

Over the years, the use of unicellular microorganisms as cell factories to obtain recombinant proteins became consolidated (Kim et al, 2015a)
Recombinant DNA techniques allow the introduction of foreign genes in a host organism for the production of heterologous proteins biologically actives
The H. polymorpha is commonly employed as an expression platform because of its unique characteristics (Figures 1A–E)

Summary

Introduction

The use of unicellular microorganisms as cell factories to obtain recombinant proteins became consolidated (Kim et al, 2015a). Recombinant DNA techniques allow the introduction of foreign genes in a host organism for the production of heterologous proteins biologically actives. Within this context, the choice of the host organism is crucial, since the functionality, solubility, and activity of the protein must be preserved during its synthesis (Vaquero et al, 2015). The H. polymorpha is commonly employed as an expression platform because of its unique characteristics (Figures 1A–E). It is thermotolerant and capable of growing at temperatures ranging from 30 to 50◦C (Figure 1B).

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