Recombinant protein expression in Pichia pastoris

The methylotrophic yeast Pichia pastoris is now one of the standard tools used in molecular biology for the generation of recombinant protein. P. pastoris has demonstrated its most powerful success as a large-scale (fermentation) recombinant protein production tool. What began more than 20 years ago as a program to convert abundant methanol to a protein source for animal feed has been developed into what is today two important biological tools: a model eukaryote used in cell biology research and a recombinant protein production system. To date well over 200 heterologous proteins have been expressed in P. pastoris. Significant advances in the development of new strains and vectors, improved techniques, and the commercial availability of these tools coupled with a better understanding of the biology of Pichia species have led to this microbe’s value and power in commercial and research labs alike.

[1]  S. Bottomley,et al.  Recombinant caspase-3 expressed in Pichia pastoris is fully activated and kinetically indistinguishable from the native enzyme. , 1997, Biochemical and biophysical research communications.

[2]  A. Beauvais,et al.  Dipeptidyl-peptidase IV secreted by Aspergillus fumigatus, a fungus pathogenic to humans , 1997, Infection and immunity.

[3]  R. Knabb,et al.  Production and purification of recombinant hirudin expressed in the methylotrophic yeast Pichia pastoris. , 1996, Protein expression and purification.

[4]  G. Sen,et al.  Synthesis and Cleavage- Secretion of Enzymatically Active Rabbit Angiotensin-converting Enzyme in Pichia pastoris* , 1996, Journal of Biological Chemistry.

[5]  High-level production of spinach glycolate oxidase in the methylotrophic yeast Pichia pastoris: engineering a biocatalyst. , 1995, Gene.

[6]  S. Sideris,et al.  Expression and characterization of recombinant mast cell tryptase. , 1999, Protein expression and purification.

[7]  S. Wagner,et al.  cDNA cloning of the 43-kDa latex allergen Hev b 7 with sequence similarity to patatins and its expression in the yeast Pichia pastoris. , 1998, European journal of biochemistry.

[8]  E. Komives,et al.  Characterization of the extracellular ligand-binding domain of the type II activin receptor. , 1998, Biochemistry.

[9]  R. F. Manning,et al.  The dacA gene of Bacillus stearothermophilus coding for D-alanine carboxypeptidase: cloning, structure and expression in Escherichia coli and Pichia pastoris. , 1993, Gene.

[10]  R. Beavis,et al.  Structural studies of alpha-N-acetylgalactosaminidase: effect of glycosylation on the level of expression, secretion efficiency, and enzyme activity. , 1998, Archives of biochemistry and biophysics.

[11]  W. Gärtner,et al.  Chromophore incorporation, Pr to Pfr kinetics, and Pfr thermal reversion of recombinant N-terminal fragments of phytochrome A and B chromoproteins. , 1998, Biochemistry.

[12]  W. Keller,et al.  Two forms of human double-stranded RNA-specific editase 1 (hRED1) generated by the insertion of an Alu cassette. , 1997, RNA.

[13]  R. Siegel,et al.  Continuous Production of a Novel Lysozyme via Secretion from the Yeast, Pichia pastoris , 1989, Bio/Technology.

[14]  A. Holder,et al.  Solution structure of an EGF module pair from the Plasmodium falciparum merozoite surface protein 1. , 1999, Journal of molecular biology.

[15]  S. Nagata,et al.  Lethal effect of recombinant human Fas ligand in mice pretreated with Propionibacterium acnes. , 1997, Journal of immunology.

[16]  J. Shiloach,et al.  Zinc-binding of endostatin is essential for its antiangiogenic activity. , 1998, Biochemical and biophysical research communications.

[17]  A. Milon,et al.  Expression and pharmacological characterization of the human μ‐opioid receptor in the methylotrophic yeast Pichia pastoris , 1996, FEBS letters.

[18]  Y. Han,et al.  Role of glycosylation in the functional expression of an Aspergillus niger phytase (phyA) in Pichia pastoris. , 1999, Archives of biochemistry and biophysics.

[19]  I. Campbell,et al.  NMR studies of a viral protein that mimics the regulators of complement activation. , 1997, Journal of molecular biology.

[20]  E. Springman,et al.  Crystal structure of recombinant human tissue kallikrein at 2.0 Å resolution , 1998, Protein science : a publication of the Protein Society.

[21]  E. Vitters,et al.  Expression of Recombinant Human Soluble Type II Transforming Growth Factor-β Receptor inPichia pastorisandEscherichia coli:Two Powerful Systems to Express a Potent Inhibitor of Transforming Growth Factor-β , 1998 .

[22]  P. Tucker,et al.  NonO enhances the association of many DNA-binding proteins to their targets. , 1997, Nucleic acids research.

[23]  M. Chapman,et al.  High-level expression of cockroach allergen, Bla g 4, in Pichia pastoris. , 1998, The Journal of allergy and clinical immunology.

[24]  F. Castellino,et al.  Glycosylationproperties of the Pichia pastoris‐expressed recombinant kringle 2 domain oftissue‐type plasminogen activator , 1997, Biotechnology and applied biochemistry.

[25]  A. Karande,et al.  Cloning, expression, purification, and immunocharacterization of placental protein-14. , 1998, Protein expression and purification.

[26]  H. Bussey,et al.  Ktr1p is an alpha-1,2-mannosyltransferase of Saccharomyces cerevisiae. Comparison of the enzymic properties of soluble recombinant Ktr1p and Kre2p/Mnt1p produced in Pichia pastoris. , 1997, The Biochemical journal.

[27]  H. Vijay,et al.  Isolation and expression of a cDNA clone encoding an Alternaria alternata Alt a 1 subunit. , 1996, International archives of allergy and immunology.

[28]  F. Döring,et al.  Expression and functional characterization of the mammalian intestinal peptide transporter PepT1 in the methylotropic yeast Pichia pastoris. , 1997, Biochemical and biophysical research communications.

[29]  P. Smith,et al.  Cloning and expression in yeast Pichia pastoris of a biologically active form of Cyn d 1, the major allergen of Bermuda grass pollen. , 1996, The Journal of allergy and clinical immunology.

[30]  S. Wu,et al.  Yeast-secreted bovine herpesvirus type 1 glycoprotein D has authentic conformational structure and immunogenicity. , 1997, Vaccine.

[31]  V. Storme,et al.  High–Level Secretion and Very Efficient Isotopic Labeling of Tick Anticoagulant Peptide (TAP) Expressed in the Methylotrophic Yeast, Pichia pastoris , 1994, Bio/Technology.

[32]  H. C. Lee,et al.  Large-scale production of human CD38 in yeast by fermentation. , 1997, Methods in enzymology.

[33]  M. Bianchi,et al.  Recombinant HMG1 protein produced in Pichia pastoris: a nonviral gene delivery agent. , 1997, BioTechniques.

[34]  C. Masters,et al.  Subcellular localization of the Alzheimer's disease amyloid precursor protein and derived polypeptides expressed in a recombinant yeast system. , 1998, Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis.

[35]  E. Komives,et al.  Large-scale expression, purification and characterization of small fragments of thrombomodulin: the roles of the sixth domain and of methionine 388. , 1995, Protein engineering.

[36]  E. Purisima,et al.  Human cathepsin X: A cysteine protease with unique carboxypeptidase activity. , 1999, Biochemistry.

[37]  G. Opdenakker,et al.  Production and characterization of recombinant active mouse gelatinase B from eukaryotic cells and in vivo effects after intravenous administration. , 1997, European journal of biochemistry.

[38]  M. Romanos.  Advances in the use of Pichia pastoris for high-level gene expression , 1995 .

[39]  R. MacGillivray,et al.  X-ray crystallography and mass spectroscopy reveal that the N-lobe of human transferrin expressed in Pichia pastoris is folded correctly but is glycosylated on serine-32. , 1999, Biochemistry.

[40]  C. Masson,et al.  Cloning and Expression of a Queen Pheromone-Binding Protein in the Honeybee: an Olfactory-Specific, Developmentally Regulated Protein , 1999, The Journal of Neuroscience.

[41]  J. de la Fuente,et al.  Effect of particulation on the immunogenic and protective properties of the recombinant Bm86 antigen expressed in Pichia pastoris. , 1998, Vaccine.

[42]  Y. Han,et al.  Cloning, sequencing, and expression of an Escherichia coli acid phosphatase/phytase gene (appA2) isolated from pig colon. , 1999, Biochemical and biophysical research communications.

[43]  P. Gros,et al.  High-level expression of mouse Mdr3 P-glycoprotein in yeast Pichia pastoris and characterization of ATPase activity. , 1998, Methods in enzymology.

[44]  N. Krishna,et al.  Expression, purification, and characterization of interferon-tau produced in Pichia pastoris grown in a minimal medium. , 1999, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[45]  B. Gerstmayer,et al.  Costimulation of T cell proliferation by a chimeric B7-2 antibody fusion protein specifically targeted to cells expressing the erbB2 proto-oncogene. , 1997, Journal of immunology.

[46]  J. G. Zhang,et al.  Evidence for the formation of a heterotrimeric complex of leukaemia inhibitory factor with its receptor subunits in solution. , 1997, The Biochemical journal.

[47]  K. Kivirikko,et al.  Assembly of human prolyl 4‐hydroxylase and type III collagen in the yeast Pichia pastoris: formation of a stable enzyme tetramer requires coexpression with collagen and assembly of a stable collagen requires coexpression with prolyl 4‐hydroxylase , 1997, The EMBO journal.

[48]  P. Roepstorff,et al.  Overexpression, purification, and characterization of recombinant barley alpha-amylases 1 and 2 secreted by the methylotrophic yeast Pichia pastoris. , 1996 .

[49]  K. Palmu,et al.  Expression in Pichia pastorisand Purification of Aspergillus awamoriGlucoamylase Catalytic Domain , 1997 .

[50]  I. Kaloshian,et al.  Characterization of LeMir, a root-knot nematode-induced gene in tomato with an encoded product secreted from the root. , 1998, Plant physiology.

[51]  C. Hollenberg,et al.  Application of yeasts in gene expression studies: a comparison of Saccharomyces cerevisiae, Hansenula polymorpha and Kluyveromyces lactis -- a review. , 1997, Gene.

[52]  J. Cregg,et al.  Functional studies of yeast-expressed human heart muscle carnitine palmitoyltransferase I. , 1997, Archives of biochemistry and biophysics.

[53]  Luis González-Candelas,et al.  Cloning of a novel constitutively expressed pectate lyase gene pelB from Fusarium solani f. sp. pisi (Nectria haematococca, mating type VI) and characterization of the gene product expressed in Pichia pastoris , 1995, Journal of bacteriology.

[54]  B. Buddle,et al.  Physiological effects and adjuvanticity of recombinant brushtail possum TNF‐α , 1999, Immunology and cell biology.

[55]  G. Winter,et al.  Improved tumour targeting by disulphide stabilized diabodies expressed in Pichia pastoris. , 1997, Protein engineering.

[56]  J. Cregg,et al.  Functional characterization of the two alcohol oxidase genes from the yeast Pichia pastoris , 1989, Molecular and cellular biology.

[57]  J. Pronk,et al.  High-Level Expression of Plasmodium vivax Apical Membrane Antigen 1 (AMA-1) in Pichia pastoris: Strong Immunogenicity in Macaca mulatta Immunized with P. vivax AMA-1 and Adjuvant SBAS2 , 1999, Infection and Immunity.

[58]  T. Tsuchiya,et al.  Macromolecular antimicrobial glycoprotein, achacin, expressed in a methylotrophic yeast Pichia pastoris , 1999, FEBS letters.

[59]  M. Yamada,et al.  Secretion of human intracellular aspartic proteinase cathepsin E expressed in the methylotrophic yeast, Pichia pastoris and characterization of produced recombinant cathepsin E. , 1994, Biochimica et biophysica acta.

[60]  C. Lima,et al.  Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[61]  F. C. Hartman,et al.  Efficient Expression of the Gene for Spinach Phosphoribulokinase in Pichia pastoris and Utilization of the Recombinant Enzyme to Explore the Role of Regulatory Cysteinyl Residues by Site-directed Mutagenesis (*) , 1996, The Journal of Biological Chemistry.

[62]  N. Crawford,et al.  Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase. , 1996, The Plant cell.

[63]  T. Kjeldsen,et al.  Secretory expression and characterization of insulin in Pichia pastoris , 1999, Biotechnology and applied biochemistry.

[64]  M. O'reilly,et al.  A recombinant human angiostatin protein inhibits experimental primary and metastatic cancer. , 1997, Cancer research.

[65]  S. Jameel,et al.  Expression and characterization of the hepatitis E virus ORF3 protein in the methylotrophic yeast, Pichia pastoris. , 1997, Gene.

[66]  C. López-Otín,et al.  Secretion of recombinant pro- and mature fungal alpha-sarcin ribotoxin by the methylotrophic yeast Pichia pastoris: the Lys-Arg motif is required for maturation. , 1998, Protein expression and purification.

[67]  J. de la Fuente,et al.  Adjuvant and immunostimulating properties of the recombinant Bm86 protein expressed in Pichia pastoris. , 1998, Vaccine.

[68]  Expression of human interleukin-17 in Pichia pastoris: purification and characterization. , 1998, Protein expression and purification.

[69]  A. Cardin,et al.  Expression of a synthetic gene encoding the anticoagulant-antimetastatic protein ghilanten by the methylotropic yeast Pichia pastoris. , 1995, Protein expression and purification.

[70]  E. Lolis,et al.  Functional and receptor binding characterization of recombinant murine macrophage inflammatory protein 2: Sequence analysis and mutagenesis identify receptor binding epitopes , 1997, Protein science : a publication of the Protein Society.

[71]  T. Gingeras,et al.  Expression of the lacZ gene from two methanol-regulated promoters in Pichia pastoris. , 1987, Nucleic acids research.

[72]  C. Masters,et al.  Processing of the Alzheimer's disease amyloid precursor protein in Pichia pastoris: immunodetection of alpha-, beta-, and gamma-secretase products. , 1998, Biochemistry.

[73]  H. Husi,et al.  Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris. , 1998, Protein expression and purification.

[74]  S. Yokoyama,et al.  Expression and secretion of rabbit plasma cholesteryl ester transfer protein by Pichia pastoris. , 1996, Journal of lipid research.

[75]  N. Hilschmann,et al.  Heterologous overexpression of human NEFA and studies on the two EF-hand calcium-binding sites. , 1999, Biochemical and biophysical research communications.

[76]  E. Lavallie,et al.  Production of a Recombinant Bovine Enterokinase Catalytic Subunit in the Methylotrophic Yeast Pichia pastoris , 1996, Bio/Technology.

[77]  F. Castellino,et al.  Enhancement through Mutagenesis of the Binding of the Isolated Kringle 2 Domain of Human Plasminogen to ω-Amino Acid Ligands and to an Internal Sequence of a Streptococcal Surface Protein* , 1999, The Journal of Biological Chemistry.

[78]  X. Liu,et al.  Biosynthetic production of type II fish antifreeze protein: fermentation by Pichia pastoris , 1997, Applied Microbiology and Biotechnology.

[79]  E. Mehler,et al.  Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. , 1999, Biochemistry.

[80]  C. Masson,et al.  Optimization of the production of a honeybee odorant-binding protein by Pichia pastoris. , 1999, Protein expression and purification.

[81]  A. Zhu,et al.  Expression, Purification, and Characterization of Recombinant α-N-Acetylgalactosaminidase Produced in the YeastPichia pastoris , 1996 .

[82]  K. O'Donoghue,et al.  Functional expression of soluble forms of human CD38 in Escherichia coli and Pichia pastoris. , 1995, Protein expression and purification.

[83]  J. Cregg,et al.  Structural comparison of the Pichia pastoris alcohol oxidase genes , 1989, Yeast.

[84]  K. Bayer,et al.  Recombinant expression of alliin lyase from garlic (Allium sativum) in bacteria and yeasts. , 1998, Planta medica.

[85]  S. Wagner,et al.  Cloning of the Patatin–Like Latex Allergen Hev b 7, Its Expression in the Yeast Pichia pastoris and Its Immunological Characterization , 1999, International Archives of Allergy and Immunology.

[86]  W. Gärtner,et al.  Effect of chromophore exchange on the resonance Raman spectra of recombinant phytochromes , 1997, FEBS letters.

[87]  R. Oriol,et al.  Heterologous expression of an engineered truncated form of human Lewis fucosyltransferase (Fuc-TIII) by the methylotrophic yeast Pichia pastoris. , 1998, Glycobiology.

[88]  P. McKee,et al.  Characterization of wild-type and mutant alpha2-antiplasmins: fibrinolysis enhancement by reactive site mutant. , 1999, Blood.

[89]  C. Nespoulous,et al.  Overexpression in Pichia pastoris and crystallization of an elicitor protein secreted by the phytopathogenic fungus, Phytophthora cryptogea. , 1996, Protein expression and purification.

[90]  K. Ogata,et al.  A Yeast Capable of Utilizing Methanol , 1969 .

[91]  S. Withers,et al.  Cloning, mutagenesis, and structural analysis of human pancreatic α‐amylase expressed in pichia pastoris , 2008, Protein science : a publication of the Protein Society.

[92]  R. Dowling,et al.  Production and Purification of Human Fibroblast Collagenase (MMP-1) Expressed in the Methylotrophic YeastPichia pastoris , 1996 .

[93]  C. Schmidt-Dannert,et al.  Functional expression of a mammalian acetylcholinesterase in Pichia pastoris: comparison to acetylcholinesterase, expressed and reconstituted from Escherichia coli. , 1998, Biochimica et biophysica acta.

[94]  R. Tirindelli,et al.  Expression of a lipocalin in Pichia pastoris: secretion, purification and binding activity of a recombinant mouse major urinary protein , 1997, FEBS letters.

[95]  L. Steinlein,et al.  Production and purification of N-terminal half-transferrin in Pichia pastoris. , 1995, Protein expression and purification.

[96]  W. Vranken,et al.  An NMR‐based identification of peptide fragments mimicking the interactions of the cathepsin B propeptide , 1998, FEBS letters.

[97]  D. Mohanraj,et al.  Expression of biologically active human vascular endothelial growth factor in yeast. , 1995, Growth Factors.

[98]  R. Schmitz,et al.  Generation of Rabbit Monoclonal Antibody Fragments from a Combinatorial Phage Display Library and Their Production in the Yeast Pichia pastoris , 1995, Bio/Technology.

[99]  M. Gautier,et al.  High-level secretion of a wheat lipid transfer protein in Pichia pastoris. , 1998, Protein expression and purification.

[100]  K. Fujikawa,et al.  Secretory production of recombinant urokinase-type plasminogen activator-annexin V chimeras in Pichia pastoris. , 1996, Gene.

[101]  T Vernet,et al.  Role of the Occluding Loop in Cathepsin B Activity* , 1997, The Journal of Biological Chemistry.

[102]  C. Handumrongkul,et al.  Cloning and expression of Candida guilliermondii xylose reductase gene (xyl1) in Pichia pastoris , 1998, Applied Microbiology and Biotechnology.

[103]  K. Moremen,et al.  Cloning, expression, purification, and characterization of the acid alpha-mannosidase from Trypanosoma cruzi. , 1998, Glycobiology.

[104]  G. Wegner.  Emerging applications of the methylotrophic yeasts. , 1990, FEMS microbiology reviews.

[105]  J. Trant.  Functional expression of recombinant spiny dogfish shark (Squalus acanthias) cytochrome P450c17 (17 alpha-hydroxylase/C17,20-lyase) in yeast (Pichia pastoris). , 1996, Archives of biochemistry and biophysics.

[106]  J. Cregg,et al.  Recombinant protein production in an alcohol oxidase-defective strain of Pichia pastoris in fedbatch fermentations , 1997 .

[107]  Michael A. Romanos,et al.  Rapid Selection Using G418 of High Copy Number Transformants of Pichia pastoris for High–level Foreign Gene Expression , 1994, Bio/Technology.

[108]  J. Fu,et al.  Expression of the dengue virus structural proteins in Pichia pastoris leads to the generation of virus-like particles. , 1997, The Journal of general virology.

[109]  S. Choe,et al.  Three-finger toxin fold for the extracellular ligand-binding domain of the type II activin receptor serine kinase. , 1999, Nature Structural Biology.

[110]  N. Gascoigne,et al.  Antibody screening for secreted proteins expressed in Pichia pastoris. , 1996, BioTechniques.

[111]  C. Raymond,et al.  Development of the methylotrophic yeast Pichia methanolica for the expression of the 65 kilodalton isoform of human glutamate decarboxylase , 1998, Yeast.

[112]  J. Tschopp,et al.  High-Level Secretion of Glycosylated Invertase in the Methylotrophic Yeast, Pichia Pastoris , 1987, Bio/Technology.

[113]  G. Daniels,et al.  Hemagglutination inhibition of Cromer blood group antibodies with soluble recombinant decay‐accelerating factor , 1998, Transfusion.

[114]  G. Lemay,et al.  Characterization of the Nucleoside Triphosphate Phosphohydrolase and Helicase Activities of the Reovirus λ1 Protein* , 1997, The Journal of Biological Chemistry.

[115]  W. McCombie,et al.  High level expression of heterologous proteins in methylotrophic yeast Pichia pastoris , 1988, Journal of basic microbiology.

[116]  S. Wagner,et al.  High level expression, purification, and characterization of the Kunitz-type protease inhibitor domain of protease nexin-2/amyloid β-protein precursor , 1992 .

[117]  F. Klis,et al.  A new tool for studying the molecular architecture of the fungal cell wall: one-step purification of recombinant trichoderma beta-(1-6)-glucanase expressed in Pichia pastoris. , 1998, Biochimica et biophysica acta.

[118]  R. Ménard,et al.  Potency and selectivity of the cathepsin L propeptide as an inhibitor of cysteine proteases. , 1996, Biochemistry.

[119]  F. Castellino,et al.  High‐level secretion in Pichia pastoris and biochemical characterization of the recombinantkringle 2 domain of tissue‐type plasminogen activator , 1997, Biotechnology and applied biochemistry.

[120]  R. Ménard,et al.  Autocatalytic Processing of Recombinant Human Procathepsin L , 1998, The Journal of Biological Chemistry.

[121]  C. Masters,et al.  Expression and analysis of heparin‐binding regions of the amyloid precursor protein of Alzheimer's disease , 1997, FEBS letters.

[122]  N. Price,et al.  Sequencing and functional expression of the malonyl-CoA-sensitive carnitine palmitoyltransferase from Drosophila melanogaster. , 1999, The Biochemical journal.

[123]  D. B. Mccormick,et al.  Riboflavin 5′-Hydroxymethyl Oxidation , 1997, The Journal of Biological Chemistry.

[124]  J. P. van Dijken,et al.  The significance of peroxisomes in the metabolism of one-carbon compounds in yeasts. , 1983, Advances in microbial physiology.

[125]  J. Rouvinen,et al.  Molecular and crystal properties of Bos d 2, an allergenic protein of the lipocalin family. , 1998, Biochemical and biophysical research communications.

[126]  I G Charles,et al.  Recombinant Bordetella pertussis pertactin (P69) from the yeast Pichia pastoris: high-level production and immunological properties. , 1991, Vaccine.

[127]  B. Davis,et al.  NMR analysis of the N-terminal SRCR domain of human CD5: engineering of a glycoprotein for superior characteristics in NMR experiments. , 1998, Protein engineering.

[128]  G. Bergamini,et al.  Intracellular production of a major cytomegalovirus antigenic protein in the methylotrophic yeast Pichia pastoris. , 1996, Gene.

[129]  D. Uhrín,et al.  Complete assignment of 1H, 13C and 15N chemical shifts for bovine β-lactoglobulin: Secondary structure and topology of the native state is retained in a partially unfolded form , 1998, Journal of biomolecular NMR.

[130]  R. Wind,et al.  High‐yield secretion of recombinant gelatins by Pichia pastoris , 1999, Yeast.

[131]  P. Howell,et al.  Purification, crystallization and preliminary X-ray crystallographic analysis of recombinant murine Golgi mannosidase IA, a class I alpha-mannosidase involved in Asn-linked oligosaccharide maturation. , 1999, Acta crystallographica. Section D, Biological crystallography.

[132]  S. Wu,et al.  A chimeric protein comprised of bovine herpesvirus type 1 glycoprotein D and bovine interleukin-6 is secreted by yeast and possesses biological activities of both molecules. , 1999, Vaccine.

[133]  T. Ueda,et al.  High‐level expression of uniformly 15N‐labeled hen lysozyme in Pichia pastoris and identification of the site in hen lysozyme where phosphate ion binds using NMR measurements , 1999, FEBS letters.

[134]  A. R. Penheiter,et al.  Purification and characterization of a soybean root nodule phosphatase expressed in Pichia pastoris. , 1998, Protein expression and purification.

[135]  Kisiel,et al.  Expression, Purification, and Inhibitory Properties of Human Proteinase Inhibitor 8 , 1997, Biochemistry.

[136]  J. Fuente,et al.  The bovine IFN-ω1 is biologically active and secreted at high levels in the yeast Pichia pastoris , 1998 .

[137]  M. Murtaugh,et al.  High-level secretion of two antibody single chain Fv fragments by Pichia pastoris. , 1997, Journal of immunological methods.

[138]  M. Romanos,et al.  High-Level Expression of Tetanus Toxin Fragment C in Pichia Pastoris Strains Containing Multiple Tandem Integrations of the Gene , 1991, Bio/Technology.

[139]  C. Fierke,et al.  Cloning, sequencing, and recombinant expression of the porcine inhibitor of carbonic anhydrase: a novel member of the transferrin family. , 1997, Biochemistry.

[140]  Hiller Km,et al.  Cloning and expression of ragweed allergen Amb a 6. , 1998 .

[141]  K. Kobayashi,et al.  Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris. , 1998, Analytical biochemistry.

[142]  A. Bollen,et al.  The conformation of purified Toxoplasma gondii SAG1 antigen, secreted from engineered Pichia pastoris, is adequate for serorecognition and cell proliferation. , 1998, Journal of biotechnology.

[143]  F. Avilés,et al.  Mapping the Pro-region of Carboxypeptidase B by Protein Engineering , 1999, The Journal of Biological Chemistry.

[144]  Hon Cheung Lee,et al.  High-Level Expression of RecombinantAplysiaADP-Ribosyl Cyclase inPichia pastorisby Fermentation , 1997 .

[145]  J. Cregg,et al.  Production of foreign proteins in the yeast Pichia pastoris , 1995 .

[146]  R. Montesino,et al.  Efficient expression and secretion of recombinant alpha amylase in Pichia pastoris using two different signal sequences , 1994, Yeast.

[147]  L. Bedzyk,et al.  Production of synthetic spider dragline silk protein in Pichia pastoris , 1997, Applied Microbiology and Biotechnology.

[148]  William C. Raschke,et al.  Recent Advances in the Expression of Foreign Genes in Pichia pastoris , 1993, Bio/Technology.

[149]  R. Contreras,et al.  Protection of mice against a lethal influenza virus challenge after immunization with yeast-derived secreted influenza virus hemagglutinin. , 1999, European journal of biochemistry.

[150]  L. D. Ward,et al.  The N-terminus of gp130 is critical for the formation of the high-affinity interleukin-6 receptor complex. , 1999, Growth factors.

[151]  R. Salesse,et al.  High-level secretion of biologically active recombinant porcine follicle-stimulating hormone by the methylotrophic yeast Pichia pastoris. , 1998, Biochemical and biophysical research communications.

[152]  J. Cregg.  6 – EXPRESSION IN THE METHYLOTROPHIC YEAST Pichia pastoris , 1999 .

[153]  P. Bird,et al.  Production and characterization of recombinant human proteinase inhibitor 6 expressed in Pichia pastoris. , 1995, Biochimica et biophysica acta.

[154]  T. Boller,et al.  Expression of a functional barley sucrose‐fructan 6‐fructosyltransferase in the methylotrophic yeast Pichia pastoris , 1998, FEBS letters.

[155]  K. Moremen,et al.  Cloning, expression, purification, and characterization of the murine lysosomal acid alpha-mannosidase. , 1997 .

[156]  R. Y. Chow,et al.  A transforming mutation enhances the activity of the c-Kit soluble tyrosine kinase domain. , 1999, The Biochemical journal.

[157]  K Rumbold,et al.  High-level intracellular expression of hydroxynitrile lyase from the tropical rubber tree Hevea brasiliensis in microbial hosts. , 1997, Protein expression and purification.

[158]  K. Palmu,et al.  Human placental alkaline phosphatase: expression in Pichia pastoris, purification and characterization of the enzyme. , 1998, Protein expression and purification.

[159]  K. Kobayashi,et al.  Analysis of Pichia pastoris components in recombinant human serum albumin by immunological assays and by HPLC with pulsed amperometric detection. , 1998, Analytical chemistry.

[160]  D. Mozley,et al.  Large‐scale Generation of Affinity‐purified Recombinant Phytochrome Chromopeptide , 1997, Photochemistry and photobiology.

[161]  K. McNatty,et al.  Expression and secretion of a biologically active glycoprotein hormone, ovine follicle stimulating hormone, by Pichia pastoris. , 1998, Journal of molecular endocrinology.

[162]  N. Acheson,et al.  Production of active polyomavirus large T antigen in yeast Pichia pastoris. , 1997, Virus research.

[163]  J. Gatehouse,et al.  Functional phytohemagglutinin (PHA) and Galanthus nivalis agglutinin (GNA) expressed in Pichia pastoris correct N-terminal processing and secretion of heterologous proteins expressed using the PHA-E signal peptide. , 1999, European journal of biochemistry.

[164]  C. Gatz,et al.  Recombinant type A and B phytochromes from potato. Transient absorption spectroscopy. , 1997, Biochemistry.

[165]  M. Lowe,et al.  Human pancreatic triglyceride lipase expressed in yeast cells: purification and characterization. , 1998, Protein expression and purification.

[166]  R. Dighe,et al.  Hyperexpression of biologically active human chorionic gonadotropin using the methylotropic yeast, Pichia pastoris. , 1999, Journal of molecular endocrinology.

[167]  J. Whisstock,et al.  Expression and purification of recombinant human granzyme B from Pichia pastoris. , 1999, Biochemical and biophysical research communications.

[168]  M. Penttilä,et al.  Laccase from the white-rot fungus Trametes versicolor: cDNA cloning of lcc1 and expression in Pichia pastoris , 1997, Current Genetics.

[169]  J. Shiloach,et al.  Disruption of the KEX1 gene in Pichia pastoris allows expression of full‐length murine and human endostatin , 1999, Yeast.

[170]  G. Lemay,et al.  A glycosyl hydrolase activity of mammalian reovirus sigma1 protein can contribute to viral infection through a mucus layer. , 1999, Journal of molecular biology.

[171]  Crystal structure of a recombinant αEC domain from human fibrinogen-420 , 1998 .

[172]  Y. Imai,et al.  Expression, purification, and neurotrophic activity of amyloid precursor protein-secreted forms produced by yeast. , 1995, Biochemical and biophysical research communications.

[173]  F. Castellino,et al.  Glycosylation of Asparagine-28 of Recombinant Staphylokinase with High-Mannose-type Oligosaccharides Results in a Protein with Highly Attenuated Plasminogen Activator Activity* , 1999, The Journal of Biological Chemistry.

[174]  J. Heyman,et al.  Labeling of peroxisomes with green fluorescent protein in living P. pastoris cells. , 1996, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[175]  Salvador Ventura,et al.  Overexpression of Human Procarboxypeptidase A2 in Pichia pastoris and Detailed Characterization of Its Activation Pathway* , 1998, The Journal of Biological Chemistry.

[176]  A. Subramaniam,et al.  Strategies for optimal synthesis and secretion of heterologous proteins in the methylotrophic yeast Pichia pastoris. , 1997, Gene.

[177]  P. Roepstorff,et al.  Processing, disulfide pattern, and biological activity of a sugar beet defensin, AX2, expressed in Pichia pastoris. , 1999, Protein expression and purification.

[178]  W. Durante,et al.  Platelet-derived Growth Factor Regulates Vascular Smooth Muscle Cell Proliferation by Inducing Cationic Amino Acid Transporter Gene Expression (*) , 1996, The Journal of Biological Chemistry.

[179]  E. Komives,et al.  Production of large quantities of isotopically labeled protein in Pichia pastoris by fermentation , 1999, Journal of biomolecular NMR.

[180]  J. Feix,et al.  Structural investigations of a new familial dysalbuminemic hyperthyroxinemia genotype. , 1999, Clinical chemistry.

[181]  A. Zhu,et al.  High-level expression and purification of coffee bean alpha-galactosidase produced in the yeast Pichia pastoris. , 1995, Archives of biochemistry and biophysics.

[182]  L. Marnett,et al.  Expression, purification, and characterization of porcine leukocyte 12-lipoxygenase produced in the methylotrophic yeast, Pichia pastoris. , 1994, Biochemical and biophysical research communications.

[183]  C. Masters,et al.  Recombinant human amyloid precursor‐like protein 2 (APLP2) expressed in the yeast Pichia pastoris can stimulate neurite outgrowth , 1999, FEBS letters.

[184]  W. Gronwald,et al.  The solution structure of type II antifreeze protein reveals a new member of the lectin family. , 1998, Biochemistry.

[185]  M. Famulok,et al.  Overexpression of active Syrian golden hamster prion protein PrPc as a glutathione S-transferase fusion in heterologous systems , 1995, Journal of virology.

[186]  H. Weinstein,et al.  Characterization of novel cathepsin K mutations in the pro and mature polypeptide regions causing pycnodysostosis. , 1999, The Journal of clinical investigation.

[187]  T. Jeffries,et al.  A strong nitrogen source-regulated promoter for controlled expression of foreign genes in the yeast Pichia pastoris. , 1998, Gene.

[188]  S. Padmanabhan,et al.  Expression of human gelatinase B in Pichia pastoris. , 1999, Protein expression and purification.

[189]  V. Berezin,et al.  The First Immunoglobulin-like Neural Cell Adhesion Molecule (NCAM) Domain Is Involved in Double-reciprocal Interaction with the Second Immunoglobulin-like NCAM Domain and in Heparin Binding* , 1997, The Journal of Biological Chemistry.

[190]  P. Gros,et al.  Mutations in either nucleotide-binding site of P-glycoprotein (Mdr3) prevent vanadate trapping of nucleotide at both sites. , 1998, Biochemistry.

[191]  G. Brudvig,et al.  Characterization of the Reductase Domain of Rat Neuronal Nitric Oxide Synthase Generated in the Methylotrophic Yeast Pichia pastoris , 1996, The Journal of Biological Chemistry.

[192]  T. Gingeras,et al.  Isolation of alcohol oxidase and two other methanol regulatable genes from the yeast Pichia pastoris , 1985, Molecular and cellular biology.

[193]  R. Stroud,et al.  Engineering a soluble extracellular erythropoietin receptor (EPObp) in Pichia pastoris to eliminate microheterogeneity, and its complex with erythropoietin. , 1999, Protein engineering.

[194]  J. Kay,et al.  Processing, Activity, and Inhibition of Recombinant Cyprosin, an Aspartic Proteinase from Cardoon (Cynara cardunculus)* , 1999, The Journal of Biological Chemistry.

[195]  C. J. Beall,et al.  Expression of human monocyte chemoattractant protein-1 in the yeast Pichia pastoris. , 1998, Protein expression and purification.

[196]  B. Szente,et al.  Human IFNγ Receptor Cytoplasmic Domain: Expression and Interaction with HuIFNγ , 1998 .

[197]  P. Stougaard,et al.  Hexose Oxidase from the Red Alga Chondrus crispus , 1997, The Journal of Biological Chemistry.

[198]  A. Wu,et al.  Expression, purification, and characterization of a two domain carcinoembryonic antigen minigene (N-A3) in pichia pastoris. The essential role of the N-domain. , 1998, Anticancer research.

[199]  F. Döring,et al.  Expression of the mammalian renal peptide transporter PEPT2 in the yeast Pichia pastoris and applications of the yeast system for functional analysis. , 1998, Molecular membrane biology.

[200]  B. Martin,et al.  Identification and Cloning of Prs a 1, a 32-kDa Endochitinase and Major Allergen of Avocado, and Its Expression in the Yeast Pichia pastoris * , 1998, The Journal of Biological Chemistry.

[201]  M. Mcgrath,et al.  Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex , 1997, Protein science : a publication of the Protein Society.

[202]  F. Castellino,et al.  Characterization of the acidic oligosaccharides assembled on the Pichia pastoris‐expressed recombinant kringle 2 domain of human tissue‐type plasminogen activator , 1997, Biotechnology and applied biochemistry.

[203]  J. de la Fuente,et al.  Large-scale production in Pichia pastoris of the recombinant vaccine Gavac against cattle tick. , 1997, Vaccine.

[204]  D. Crawford,et al.  Cloning of clustered Streptomyces viridosporus T7A lignocellulose catabolism genes encoding peroxidase and endoglucanase and their extracellular expression in Pichia pastoris. , 1998, Canadian journal of microbiology.

[205]  J J Clare,et al.  Production of mouse epidermal growth factor in yeast: high-level secretion using Pichia pastoris strains containing multiple gene copies. , 1991, Gene.

[206]  N. Crawford,et al.  Analysis of Wild-Type and Mutant Plant Nitrate Reductase Expressed in the Methylotrophic Yeast Pichia pastoris , 1997, Plant physiology.

[207]  M. Selkirk,et al.  Cloning, Expression, and Properties of a Nonneuronal Secreted Acetylcholinesterase from the Parasitic Nematode Nippostrongylus brasiliensis * , 1999, The Journal of Biological Chemistry.

[208]  K. Okabayashi,et al.  Secretion of a variant of human single‐chain urokinase‐type plasminogen activator without an N‐glycosylation site in the methylotrophic yeast, Pichia pastoris and characterization of the secreted product , 1996, Yeast.

[209]  J. Hiltunen,et al.  Peroxisomal multifunctional enzyme of beta-oxidation metabolizing D-3-hydroxyacyl-CoA esters in rat liver: molecular cloning, expression and characterization. , 1997, The Biochemical journal.

[210]  L. Smith.  Development of recombinant vaccines for botulinum neurotoxin. , 1998, Toxicon : official journal of the International Society on Toxinology.

[211]  D. Johnson,et al.  Recombinant human mast cell tryptase beta: stable expression in Pichia pastoris and purification of fully active enzyme , 1998, Biotechnology and applied biochemistry.

[212]  R. Potenz,et al.  High-level expression, purification, and characterization of recombinant human tumor necrosis factor synthesized in the methylotrophic yeast Pichia pastoris. , 1989, Biochemistry.

[213]  P. Gros,et al.  Mutations in the nucleotide-binding sites of P-glycoprotein that affect substrate specificity modulate substrate-induced adenosine triphosphatase activity. , 1998, Biochemistry.

[214]  M. Kirkitadze,et al.  Independently melting modules and highly structured intermodular junctions within complement receptor type 1. , 1999, Biochemistry.

[215]  T. P. King,et al.  Expressions of recombinant venom allergen, antigen 5 of yellowjacket (Vespula vulgaris) and paper wasp (Polistes annularis), in bacteria or yeast. , 1999, Protein expression and purification.

[216]  N. Morel,et al.  Expression and processing of vertebrate acetylcholinesterase in the yeast Pichia pastoris. , 1997, The Biochemical journal.

[217]  L. Keegan,et al.  Purification of native and recombinant double-stranded RNA-specific adenosine deaminases. , 1998, Methods.

[218]  R. Ménard,et al.  Synthesis of amidrazones using an engineered papain nitrile hydratase 1 , 1998, FEBS letters.

[219]  C. Scorer,et al.  The intracellular production and secretion of HIV-1 envelope protein in the methylotrophic yeast Pichia pastoris. , 1993, Gene.

[220]  R. Contreras,et al.  Protection of mice against a lethal influenza challenge by immunization with yeast-derived recombinant influenza neuraminidase. , 1997, European journal of biochemistry.

[221]  M. Schlaak,et al.  Comparison of natural and recombinant isoforms of grass pollen allergens , 1997, Electrophoresis.

[222]  R. Armitage,et al.  Expression of trimeric CD40 ligand in Pichia pastoris: use of a rapid method to detect high-level expressing transformants. , 1997, Gene.

[223]  M. Popp,et al.  Functional expression of bovine opsin in the methylotrophic yeast Pichia pastoris. , 1997, Protein expression and purification.

[224]  K. Beyreuther,et al.  Expression of human amyloid precursor protein ectodomains in Pichia pastoris: analysis of culture conditions, purification, and characterization. , 1997, Protein expression and purification.

[225]  R. Brierley,et al.  Secretion of recombinant human insulin-like growth factor I (IGF-I). , 1998, Methods in molecular biology.

[226]  R. Dighe,et al.  Expression of biologically active beta subunit of bovine follicle-stimulating hormone in the methylotrophic yeast Pichia pastoris. , 1997, Protein expression and purification.

[227]  J. Strominger,et al.  Expression of Recombinant HLA-DR2 Molecules , 1996, The Journal of Biological Chemistry.

[228]  C. Raymond.  7 – RECOMBINANT PROTEIN EXPRESSION IN Pichia methanolica , 1999 .

[229]  M. Penichet,et al.  High level expression of the B. microplus Bm86 antigen in the yeast Pichia pastoris forming highly immunogenic particles for cattle. , 1994, Journal of biotechnology.

[230]  P. Pétra,et al.  Heterologous expression of wild type and deglycosylated human sex steroid-binding protein (SBP or SHBG) in the yeast, Pichia pastoris. Characterization of the recombinant proteins , 1999, The Journal of Steroid Biochemistry and Molecular Biology.

[231]  R. O. Morris,et al.  Isolation of a gene encoding a glycosylated cytokinin oxidase from maize. , 1999, Biochemical and biophysical research communications.

[232]  P. Eldin,et al.  In vitro production of recombinant antibody fragments in Pichia pastoris. , 1998, Research in immunology.

[233]  D. Tessier,et al.  High-Level Production of RecombinantGeotrichum candidumLipases in YeastPichia pastoris , 1997 .

[234]  B. Chung,et al.  Glycosylation of human α1‐antitrypsin in Saccharomyces cerevisiae and methylotrophic yeasts , 1998 .

[235]  F. Soubrier,et al.  Drosophila melanogaster angiotensin I-converting enzyme expressed in Pichia pastoris resembles the C domain of the mammalian homologue and does not require glycosylation for secretion and enzymic activity. , 1996, The Biochemical journal.

[236]  C. Scorer,et al.  Foreign gene expression in yeast: a review , 1992, Yeast.

[237]  J. Cregg,et al.  Heterologous protein expression in the methylotrophic yeast Pichia pastoris. , 2000, FEMS microbiology reviews.

[238]  J. Kaper,et al.  Production of Vibrio cholerae accessory cholera enterotoxin (Ace) in the yeast Pichia pastoris , 1997, Infection and immunity.

[239]  M. Schlaak,et al.  Grass group I allergens (β‐expansins) are novel, papain‐relatedproteinases , 1999 .

[240]  R. MacGillivray,et al.  Production and isolation of the recombinant N-lobe of human serum transferrin from the methylotrophic yeast Pichia pastoris. , 1996, Protein expression and purification.

[241]  M. Eckart,et al.  Quality and authenticity of heterologous proteins synthesized in yeast. , 1996, Current opinion in biotechnology.

[242]  J. Cregg,et al.  High–Level Expression and Efficient Assembly of Hepatitis B Surface Antigen in the Methylotrophic Yeast, Pichia Pastoris , 1987, Bio/Technology.

[243]  A. Martínez-Ruíz,et al.  Production and detailed characterization of biologically active olive pollen allergen Ole e 1 secreted by the yeast Pichia pastoris. , 1999, European journal of biochemistry.

[244]  Leonard A. Smith,et al.  Purification, Potency, and Efficacy of the Botulinum Neurotoxin Type A Binding Domain from Pichia pastoris as a Recombinant Vaccine Candidate , 1998, Infection and Immunity.

[245]  J. Almond,et al.  Interaction between echovirus 7 and its receptor, decay-accelerating factor (CD55): evidence for a secondary cellular factor in A-particle formation , 1997 .

[246]  J. Cregg,et al.  Development of yeast transformation systems and construction of methanol-utilization-defective mutants of Pichia pastori by gene disruption , 1987 .

[247]  S. Solapure,et al.  Production of Recombinant Human Bile Salt Stimulated Lipase and Its Variant inPichia pastoris , 1998 .

[248]  K. A. Parker,et al.  Expression of streptokinase in Pichia pastoris yeast , 1989 .

[249]  D. Wishart,et al.  Vl-linker-Vh orientation-dependent expression of single chain Fv-containing an engineered disulfide-stabilized bond in the framework regions. , 1995, Journal of biochemistry.

[250]  F. Castellino,et al.  O‐Mannosylation of Pichia pastoris cellular and recombinant proteins , 1998, Biotechnology and applied biochemistry.

[251]  P. Heeringa,et al.  Recombinant proteinase 3 (Wegener's antigen) expressed in Pichia pastoris is functionally active and is recognized by patient sera , 1997, Clinical and experimental immunology.

[252]  Han,et al.  Sequence polymorphism of the group 1 allergen of Bermuda grass pollen , 1999, Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology.

[253]  G. Casadoro,et al.  Secretion, purification and activity of two recombinant pepper endo‐β‐1,4‐glucanases expressed in the yeast Pichia pastoris , 1998, FEBS letters.

[254]  C. J. Beall,et al.  Inhibition ofAspergillusSerine Proteinase byStreptomycesSubtilisin Inhibitor and High-Level Expression of This Inhibitor inPichia pastoris , 1996 .

[255]  A. Herscovics,et al.  Molecular cloning, chromosomal mapping and tissue-specific expression of a novel human alpha1,2-mannosidase gene involved in N-glycan maturation. , 1998, Glycobiology.

[256]  P. Andreasen,et al.  Binding of urokinase-type plasminogen activator-plasminogen activator inhibitor-1 complex to the endocytosis receptors alpha2-macroglobulin receptor/low-density lipoprotein receptor-related protein and very-low-density lipoprotein receptor involves basic residues in the inhibitor. , 1998, The Biochemical journal.

[257]  M. Schliwa,et al.  Expression of Reticulomyxa filosa tubulins in Pichia pastoris: regulation of tubulin pools , 1997, FEBS letters.

[258]  E. Davie,et al.  The Polymerization Pocket “a” within the Carboxyl-terminal Region of the γ Chain of Human Fibrinogen Is Adjacent to but Independent from the Calcium-binding Site* , 1997, The Journal of Biological Chemistry.

[259]  R. Montesino,et al.  Variation in N-linked oligosaccharide structures on heterologous proteins secreted by the methylotrophic yeast Pichia pastoris. , 1998, Protein expression and purification.

[260]  C. Schmidt-Dannert,et al.  Functional expression of Rhizopus oryzae lipase in Pichia pastoris: high-level production and some properties. , 1998, Journal of biotechnology.

[261]  D. Shotton,et al.  Structural analysis of the CD5 antigen--expression, disulphide bond analysis and physical characterisation of CD5 scavenger receptor superfamily domain 1. , 1998, European journal of biochemistry.

[262]  J. Latgé,et al.  Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus , 1997, Infection and immunity.

[263]  H. Michel,et al.  Comparative biochemical and pharmacological characterization of the mouse 5HT5A 5-hydroxytryptamine receptor and the human beta2-adrenergic receptor produced in the methylotrophic yeast Pichia pastoris. , 1998, The Biochemical journal.

[264]  M. Payne,et al.  Engineering Pichia pastoris for biocatalysis: co-production of two active enzymes. , 1997, Gene.

[265]  M. Ho,et al.  Altered antigenicity of 'a' determinant variants of hepatitis B virus. , 1997, The Journal of general virology.

[266]  K. Moremen,et al.  Cloning, Expression, Purification, and Characterization of the Human Broad Specificity Lysosomal Acid α-Mannosidase* , 1996, The Journal of Biological Chemistry.

[267]  F. Maley,et al.  Structure of oligosaccharides on Saccharomyces SUC2 invertase secreted by the methylotrophic yeast Pichia pastoris. , 1991, The Journal of biological chemistry.

[268]  D. Luo,et al.  Expression of a fusion protein of scFv-biotin mimetic peptide for immunoassay. , 1998, Journal of biotechnology.

[269]  J. Alexander,et al.  Production of the rat complement regulator, Crry, as an active soluble protein in Pichia pastoris. , 1997, Archives of biochemistry and biophysics.

[270]  R. Jiménez-Flores,et al.  Study of Putative Glycosylation Sites in Bovine β-Casein Introduced by PCR-Based Site-Directed Mutagenesis , 1996 .

[271]  A. Kaplan,et al.  Atypical processing of amyloid precursor fusion protein by proteolytic activity in Pichia pastoris. , 1999, Biochemical and biophysical research communications.

[272]  N. Gonatas,et al.  The Golgi sialoglycoprotein MG160, expressed in Pichia pastoris, does not require complex carbohydrates and sialic acid for secretion and basic fibroblast growth factor binding. , 1997, Biochemical and biophysical research communications.

[273]  E. Springman,et al.  Expression and characterization of human tissue kallikrein variants. , 1998, Protein expression and purification.

[274]  A. Beauvais,et al.  Biochemical and Antigenic Characterization of a New Dipeptidyl-Peptidase Isolated from Aspergillus fumigatus* , 1997, The Journal of Biological Chemistry.

[275]  M. Lowe,et al.  Purification and characterization of human procolipase expressed in yeast cells. , 1998, Protein expression and purification.

[276]  D. Higgins.  Overview of Protein Expression in Pichia pastoris , 1995, Current protocols in protein science.

[277]  Expression, crystallization and preliminary X-ray analysis of the two amino-terminal Ig domains of the neural cell adhesion molecule (NCAM). , 1999, Acta crystallographica. Section D, Biological crystallography.

[278]  G. Opdenakker,et al.  Expression of a human mutant monocyte chemotactic protein 3 in Pichia pastoris and characterization as an MCP-3 receptor antagonist. , 1995, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[279]  K. Kragh,et al.  A group of alpha-1,4-glucan lyases and their genes from the red alga Gracilariopsis lemaneiformis: purification, cloning, and heterologous expression. , 1999, Biochimica et biophysica acta.

[280]  M. Hirose,et al.  Complete determination of disulfide forms of purified recombinant human serum albumin, secreted by the yeast Pichia pastoris. , 1997, Analytical chemistry.

[281]  J. Baratti,et al.  Oxidation of Methanol by the Yeast, Pichia pastoris. Purification and Properties of the Alcohol Oxidase , 1980 .

[282]  J. Cregg,et al.  Reconstitution of highly expressed human heart muscle carnitine palmitoyltransferase I. , 1997, Biochemical and biophysical research communications.

[283]  P. Roepstorff,et al.  Overexpression and characterization of Aspergillus awamori wild-type and mutant glucoamylase secreted by the methylotrophic yeast Pichia pastoris: comparison with wild-type recombinant glucoamylase produced using Saccharomyces cerevisiae and Aspergillus niger as hosts. , 1997, Protein expression and purification.

[284]  H. Waterham,et al.  Isolation of the Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase gene and regulation and use of its promoter. , 1997, Gene.

[285]  J. Massoulie,et al.  Cloning and expression of acetylcholinesterase from Electrophorus. Splicing pattern of the 3' exons in vivo and in transfected mammalian cells. , 1997, The Journal of biological chemistry.

[286]  Luis González-Candelas,et al.  Identification of a novel pelD gene expressed uniquely in planta by Fusarium solani f. sp. pisi (Nectria haematococca, mating type VI) and characterization of its protein product as an endo-pectate lyase. , 1996, Archives of biochemistry and biophysics.

[287]  S. Fedosov,et al.  Sequence, S–S Bridges, and Spectra of Bovine Transcobalamin Expressed in Pichia pastoris * , 1999, The Journal of Biological Chemistry.

[288]  H. Waterham,et al.  Functional characterization of mitochondrial carnitine palmitoyltransferases I and II expressed in the yeast Pichia pastoris. , 1997, Biochemistry.

[289]  C. Batt,et al.  High-level expression of bovine beta-lactoglobulin in Pichia pastoris and characterization of its physical properties. , 1997, Protein engineering.