Chapter 13 Expression in the Yeast Pichia pastoris

Cregg, James M.; Tolstorukov, Ilya; Kusari, Anasua B.; Sunga, Jay; Madden, Knut R.; Chappell, Thomas G.

doi:10.1016/s0076-6879(09)63013-5

Cited by 197 publications

(128 citation statements)

References 8 publications

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“…Methylotrophic yeast, P. pastoris, has been developed as a useful host for the highlevel expression of heterologous peptides and proteins under the AOX1 (alcohol oxidase 1) gene promoter [33], which is tightly regulated by methanol. P. pastoris have some advantages over prokaryotic cells, especially in regards to post-transcriptional and post-translational modifications, such as glycosylation [34], also the yeast allow secretion of the recombinant protein, which results in fewer purification steps, and consequently has an increased protein yield and facilitated scale-up process. Recently, some antimicrobial peptides were successfully expressed in the P. pastoris, including bovine lactoferrampin and bovine lactoferricin [35], perinerin of clamworm [25], and hPAB-␤ [36].…”

Section: Dissussionmentioning

confidence: 99%

Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system

Zhao

Tang

Cao

et al. 2015

Enzyme and Microbial Technology

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Section: Dissussionmentioning

confidence: 99%

Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system

Zhao

Tang

Cao

et al. 2015

Enzyme and Microbial Technology

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“…There are several advantages to the P. pastoris expression system, including rapid growth and convenient prokaryotic and eukaryotic post-translational processing and modification machinery (Cregg et al 2009;Damasceno et al 2012;Kang et al 2013). The P. pastoris expression system also allows for both intracellular and secretory expression of a sequence of interest.…”

Section: Discussionmentioning

confidence: 99%

Expression and purification of recombinant human neuritin from Pichia pastoris and a partial analysis of its neurobiological activity in vitro

Zhang

Xian

et al. 2015

Appl Microbiol Biotechnol

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Neuritin (also known as candidate plasticity gene 15 (cpg15)) is a neurotrophic factor that was recently discovered in a screen aimed at identifying genes involved in activity-dependent synaptic plasticity. Neuritin plays multiple roles in both neural development (Chen et al. Zhonghua Yan Ke Za Zhi 46:978-983 2010; Corriveau et al. J Neurosci 19:7999-8008 1999; Lee and Nedivi J Neurosci 22:1807-1815 2002) and synaptic plasticity (Fujino et al. Gene Dev 25:2674-2685 2011; Leslie and Nedivi Prog 14 Neurobiol 94:223-237 2011; Loebrich and Nedivi Physiol Rev 89:1079 2009). In this study, to produce bioactive, soluble recombinant human neuritin protein, a portion of NRN1 was cloned into the Pichia pastoris expression vector pPIC9K. The recombinant vector was then transformed into the methylotrophic yeast strain P. pastoris GS115, and a shaking flask method and His-tag purification strategy were utilized to express and purify neuritin protein. The resulting protein had a molecular mass of approximately 11 kDa, and subsequent functional analyses indicated that the purified neuritin promoted neurite outgrowth from embryonic chicken dorsal root ganglions, while also prolonging the survival of these ganglions, and from PC12 cells. These findings suggest that neuritin was expressed effectively in vitro and that this protein may play a role in stimulating neurite outgrowth of both dorsal root ganglions and PC12 cells. This study provides a novel strategy for the large-scale production of bioactive neuritin, which will enable further study of the biological function of this protein.

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“…To obtain the high-quality and extracellular expression of recombinant gp350, we chose pPICZαA and P. pastoris GS115 as the expression vector and host strain, respectively. However, several factors such as P. pastoris codon bias and base content of foreign genes have been proved to markedly influence the expression of heterologous genes (Cregg et al 2009). Codon optimization has often been implemented to improve the expression of heterologous genes.…”

Section: Discussionmentioning

confidence: 99%

Expression and immunogenic characterization of recombinant gp350 for developing a subunit vaccine against Epstein-Barr virus

Wang

Jiang

Han

et al. 2015

Appl Microbiol Biotechnol

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Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that is linked to the development of various malignancies. There is an urgent need for effective vaccines against EBV. EBV envelope glycoprotein gp350 is an attractive candidate for a prophylactic vaccine. This study was undertaken to produce the truncated (codons 1-443) gp350 protein (gp350(1-443)) in Pichia pastoris and evaluate its immunogenicity. The gp350(1-443) protein was expressed as a secretory protein with an N-terminal His-tag in P. pastoris and purified through Ni-NTA chromatography. Immunization with the recombinant gp350(1-443) could elicit high levels of gp350(1-443)-specific antibodies in mice. Moreover, gp350(1-443)-immunized mice developed strong lymphoproliferative and Th1/Th2 cytokine responses. Furthermore, the recombinant gp350(1-443) could stimulate CD4(+) and CD8(+) T cell responses in vaccinated mice. Collectively, these findings demonstrated that the yeast-expressed gp350(1-443) retained strong immunogenicity. This study will provide a useful source for developing EBV subunit vaccine candidates.

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Chapter 13 Expression in the Yeast Pichia pastoris

Cited by 197 publications

References 8 publications

Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system

Characterization of bioactive recombinant antimicrobial peptide parasin I fused with human lysozyme expressed in the yeast Pichia pastoris system

Expression and purification of recombinant human neuritin from Pichia pastoris and a partial analysis of its neurobiological activity in vitro

Expression and immunogenic characterization of recombinant gp350 for developing a subunit vaccine against Epstein-Barr virus

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