Expression Pichia pastoris

R)- and ( -selective HNLs. A number of recombinant HNLs have also been expressed in E. coli, Saccharomyces cerevisiae, and Pichia pastoris. Recently, protein engineering has been successfully applied to the development of a tailor-made HNL for large-scale production of specific cyanohydrins [69,70]. 

Macauley-Patrick, S., Fazenda, M.L., McNeil, B. and Harvey, L.M. (2005) Heterologous protein production using the Pichia pastoris expression system. Yeast, 22, 249-270. 

Chen, Y.-R., Huang, H.-H., Cheng, Y.-F. et al. (2006) Expression of a cholesterol oxidase gene from Arthrobacter simplex in Escherichia coli and Pichia pastoris. Enzyme and Microbial Technology, 39, 854-860. 

Guoa, M., Hang, H. and Zhua, T. (2008) Effect of glycosylation on biochemical characterization of recombinant phytase expressed in Pichia pastoris. Enzyme and Microbial Technology, 42, 340-345. 

Sletta, H., Aune, R., Nedal, A. etal. (2007) The presence of N-terminal secretion signal sequences leads to strong stimulation of the total expression levels of three tested medically important proteins during high-cell-density cultivations of Pichia pastoris. Applied and Environmental Microbiology, 73 (3), 906-912. 

Baneyx, F. I 491 Rcconibi nail protein expression in Pichia pastoris. Current Opinion in Biotechnology, 10(5), 411—421. 

Sorensen, H.P. and Mortensen, K.K. (2005) Advanced genetic strategies for recombinant protein expression in Pichia pastoris. Journal of Biotechnology, 115 (2), 113-128. 

Cereghino, J.L. and Cregg, J.M. (2000) Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiology Reviews, 24 (1), 45-66. 

Gellissen, G., Kunze, G., Gaillardin, C. et al. (2005) New yeast expression platforms based on methylotrophic Hansenula polymorpha and Pichia pastoris and on dimorphic Arxula adeninivorans and Yarrowia lipolytica - a comparison. FEMS Yeast Research, 5 (11), 1079-1096. 

Hermann, M., Kietzmann, M.U., Ivancic, M. et al. (2008) Alternative pig liver esterase (APLE) - cloning, identification and functional expression in Pichia pastoris of a versatile new biocatalyst. Journal of Biotechnology, 133 (3), 301-310. 

Doring, F., S. Theis, and H. Daniel. Expression and functional characterization of the mammalian intestinal peptide transporter PepTl in the methylotropic yeast Pichia pastoris. Biochem. Biophys. Res. Commun. 1997, 232, 656-662. 

Although proteins can be expressed in many heterologous production systems, including bacteria such as Proteus mirabilis [1], fungi such as Pichia pastoris [2, 3] and Aspergillus awamori [4] and insect cells [5, 6], the pharmaceutical industry has narrowed down process development to a small number of platform technologies ... 

ANDERSEN, M.D., M0LLER, B.L., Cytochromes P450 from Cassava (Manihot esculenta Crantz) catalyzing the first steps in the biosynthesis of the cyanogenic glucosides linamarin and lotaustralin cloning, functional expression in Pichia pastoris and substrate specificity of the isolated recombinant enzymes, J. Biol. Chem., 2000,275, 1966-1975. 

Kristensen AK, Brunstedt J, Nielsen JE, Mikkelsen JD, Roepsstorff P, Nielsen KK. Processing, disulfide pattern and biological activity of sugar beet defensin AX2, expression in Pichia pastoris. Protein Expr Purif 1999 16 377-387. 

Fig. 18.—C-l Regions of 13C-N.m.r. Spectra of a,)3-D-Mannopyranans from Pichia pastoris (A) and Citeromyces matritensis (B), and Those of Oligosaccharides Formed by Partial Acetolysis (C and D, Respectively). (Solvent, DjO temperature, 70° chemical shifts expressed as 8C, relative to external tetramethylsilane.)...

Pichia pastoris High GPCR expression levels Selection procedure required... 

Eukaryotic expression yeast Pichia pastoris] Rapid growth with ease of scale-up and Glycosylated product differs from... 

There are two yeast expression hosts that have an established track record for high-level production of heterologous proteins, namely Saccharomyces cerevisiae and Pichia pastoris. HTP expression screening using microplate formats has been reported for both these yeasts by Lang and coworkers (Holz et ah, 2002, 2003 Boettner et ah, 2002). In both cases standard protocols have been miniaturized with cells cultured in either 1.5 ml cultures in 96-deep-well plates for S. cerevisiae or 2 ml cultures in 24-deep-well plates for P. pastoris. Soluble... 

Boettner, M., Prinz, B., Holz, C., Stahl, U. and Lang, C. (2002). High-throughput screening for expression of heterologous proteins in the yeast Pichia pastoris. J. Biotechnol 99,51-62. 

ImHNL is a nonglycosylated homodimer (84 kDa) which catalyzes the reversible cleavage of aliphatic (R)-cyanohydrins [34]. This HNL does not require complex protein modification after protein biosynthesis. Thus, expression in prokaryotic Escherichia coli) and eukaryotic hosts Pichia pastoris) is possible [35-37]. However, initial trials to express IwHNL in E. coli were hampered by formation of inclusion bodies [36]. 

RK is cloned and expressed in SF9 cells [17,23,32] cDNA encoding RK is characterized and sequenced, 561 amino acids protein [22] RK gene expression in baculovirus-infected Sf21 cells [25,30] RK gene is cloned and expressed in Pichia pastoris GS115, COS-1 cells and HEK-293 stable cell line, best in COS-1 cells with correct posttranslational modifications [28] expression of RK and mutants in COS-7 cells [33]) [17, 22, 23, 25, 28, 30, 32, 33] <4> (cDNA encoding enzyme is cloned and expressed in COS-7 cells, sequence of the 561 amino acids protein [13]) [13]... 

H. Lehrach, and D. Cahill, A system for dual protein expression in Pichia pastoris, Prot. Expr. Purif. 2000, 20, 372-378. 

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