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  • Pichia pastoris Expression Strains

    BioGrammatics offers two series of Pichia pastoris expression strains. Our house strain series (BG strains) is derived from a wild Pichia pastoris strain originally isolated by Herman Phaff in California during the 1940's. Our single colony isolate of the Phaff strain was fully sequenced using Illumina HiSeq technology. This original BioGrammatics strain, BG08, was the starting point for our strain series. BG08 was grown under selective conditions to eliminate cytoplasmic killer plasmids and generate our wild type expression strain, BG10. With the exception of BG09, all subsequent strains in the BG series are derived by precise genomic manipulation of BG10. BG09 is a specific variant of BG08 that has been developed for isotopically labeling proteins with 13C at tryptic cleavage sites for mass spectroscopy studies.

    In addition to the BG series, BioGrammatics is also a distributor of RCT's GlycoSwitch® strains. The GlycoSwitch® strain series is derived from the Pichia pastoris GS115 strain developed at Phillips Petroleum in the 1970's. GS115 is a mutagenized isolate of one of the original Phaff strains, selected for histidine auxotrophy. The original GlycoSwitch® Man5 strain originates from collaborations amongst Roland Contreras, Ph.D., Nico Callewaert, Ph.D., and their colleagues at Ghent University and the Flanders Institute for Biotechnology (VIB) in Belgium. The SuperMan5 strains are subsequent modifications of GlycoSwitch® Man5 generated by BioGrammatics, in which all regions of genomic homology at the mutant och1 locus have been removed to prevent reversion to wild type by homologous recombination.

    All strains are available as YPD stabs. A selection of strains is also available as electrocompetent cells—ready for transformation.

    BioGrammatics' wild type expression strain of Pichia pastoris. This strain has been characterized by full genomic sequencing and 5000+ gene transcriptome data sets generated for various growth conditions, including shake flask and fermentation cultures. In fermenation, BioGrammatics has achieved cell densities in excess of 700 grams wet cell pellet / liter with this strain.
    An arginine, lysine auxotrophic strain developed for SILAC experiments. The genotype of BG09 is arg4Δ::nourseothricinR, lys2Δ::hygromycinR. This strain needs to be supplemented with arginine and lysine for growth. 13C arginine and/or lysine can be used to uniformly shift the molecular weights of tryptic fragments of expressed proteins.
    A slow methanol utilization deriviative of BG10. The AOX1 open reading frame has been deleted from ATG to stop. This strain grows slowly when methanol is used as the sole carbon source, and is most useful for fermenatation optimization studies.
    A histidine auxotrophic strain developed for use with older, non-BioGrammatics HIS4 expression vectors. This strain needs to be supplemented with histidine for growth. It is recommended that this strain be used only for transformation with HIS4 expression vectors. The genotype of BG12 is his4Δ, with no remaining selection marker at the his4 locus.
    Vacuolar aspartyl protease (proteinase A) deletion strain, derived from BG10. The genotype of BG20 is pep4Δ.
    Secreted subtilisin deletion strain, derived from BG10. The genotype of BG21 is sub2Δ.
    under construction
    Short answer:
    Most people using Pichia pastoris for protein expression studies are really working with Komagataella phaffii.
    Long answer:
    The genus Pichia is an old, 19th century catchall for many yeast species. It's so broad that the various yeasts in the genus don't even use the same genetic code. To further complicate the story, the original wild French Pichia pastoris isolate wasn't even placed in the Pichia genus at the time. When Herman Phaff isolated what would eventually become known as Komagataella phaffii in California, he characterized it as being identical to the French isolate and he reclassified both into the Pichia genus. With the development of DNA sequencing and PCR, it became apparent that there were actually three different but related methylotrophic species that were all being called Pichia pastoris. This clade of three was reclassified under the new methylotrophic genus Komagataella in the late 1990's. Of the three species—K. pastoris, K. phaffii and K. pseudopastoris—K. phaffii is the most commonly used heterologous protein expression host. Officially, Pichia pastoris is no longer a species name, but rather just a colloquial designation for the heterologous expression system.
    And Pichia is easier to pronounce than Komagataella...
      Product Description
      SuperMan5 (HIS+) GlycoSwitch® strain, Man5 N-linked oligosaccharide structures Details
      SuperMan5 (his) GlycoSwitch® strain, Man5 N-linked oligosaccharide structures (histidine auxotroph) Details
      Pichia pastoris BG10 strain Wild type Pichia pastoris expression strain (killer plasmid free) Details
      Pichia pastoris BG11 strain Derivative of Pichia pastoris BG10 strain, aox1Δ, MutS (methanol utilization slow) Details
      SuperMan5 (HIS+, pep4Δ) GlycoSwitch® strain, Man5 N-linked oligosaccharide structures (PEP4 protease deletion) Details
      Pichia pastoris BG09 strain Derivative of Pichia pastoris BG08 strain, arg4Δ::nourseothricinR, lys2Δ::hygromycinR (arginine, lysine auxotroph) Details
      Pichia pastoris BG12 strain Derivative of Pichia pastoris BG10 strain, his4Δ (histidine auxotroph) Details
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