What are the common vector/plasmids used in the Pichia pastoris Protein expression system?

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The expression of any recombinant gene in P. pastoris has three phase: (a) Cloning of a new gene into a suitable expression vector, (b) insertion of the cloned vector into P. pastoris host genome; and (c) trial of the potential different strains for the expression of the recombinant integrated gene.

What are the major components in the expression vector of P. pastoris?

The expression vectors in the P. pastoris expression system are one of the major components of this system. These vectors are composed of three sequences:

  • Promoter sequence (most often AOX1) in 5′ region
  • Transcriptional termination sequence in 3′ region which is essential in the processing and polyadenylation of messenger RNAs
  • Single or multiple cloning sites, necessary for the insertion of the gene of interest.

Why expression vector of Pichia pastoris need to be linearized?

The episomal vectors can replicate either autonomously in the cytoplasm or as part of a chromosome. But the vectors used in P. pastoris have no stable episomal status; therefore, they should first be linearized with enzymes and then be integrated into the P. pastoris chromosome.

Common plasmids used in the P. pastoris expression system to produce extracellular and intracellular proteins are listed in Tables 1 and 2, respectively.

Vector nameMarker geneUsed strainRecombinant protein
pPIC3.5KHis4, Kan, AmpKM71Maltooligosyltrehalose synthase
SMD1168Camellia sinensisÊheat shock protein
GS115Pleurotus ostreatusÊlaccases
GS115Rhizopus oryzaeÊLipase
GS115HSA/GH fusion protein
KM71Membrane protein
KM71Dengue virus envelope glycoprotein
pHIL_D2His4, AmpGS115Prostaglandin H synthase_2
GS115CatA1 and SODC
KM71RhodococcusÊnitrile hydratase

Table 1. Common Pichia pastoris expression vectors for the production of intracellular proteins

Vector nameMarker geneUsed strainRecombinant protein
pPIC9KHis4, Kan, AmpGS115Xylanase
GS115Porcine circovirus type 2
pPICZ_ShbleSMD1168Human chitinase
GS115Human topoisomerase I
GS115Human interferon gamma
X_33C_reactive protein
X_33Human RNase4
pHIL_S1His4, AmpGS115Rabies virus glycoprotein
GS115Rhizopus oryzaeÊLipase
KM71Camel lactoferricin
pGAPZ_ShbleGS115Acyl homoserine lactonase
SMD1168Variable lymphocyte receptor B
X_33Human gastric lipase
pJL_SXFLD1, AmpMS105Formaldehyde dehydrogenase
pBLHIS_SXHis4, AmpJC100Leukocyte protease inhibitor

Table 2. Common Pichia pastoris expression vectors for the production of secretory proteins

Ref: Pichia pastoris: A highly successful expression system for optimal synthesis of heterologous proteins

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