High level expression of heterologous proteins in methylotrophic yeast Pichia pastoris - PubMed (original) (raw)
High level expression of heterologous proteins in methylotrophic yeast Pichia pastoris
K Sreekrishna et al. J Basic Microbiol. 1988.
Abstract
Expression of human tumor necrosis factor-alpha (TNF) and four different TNF analogs has been studied in Pichia pastoris by utilizing the alcohol oxidase gene promoter. TNF expression level in certain transformants accounted for as much as 36% of the soluble protein. TNF expression was stably maintained during high cell density fermentation (100 g dry cell weight/liter) resulting in a TNF production level of 6-10 g/liter. TNF contained in P. pastoris cell lysates was biologically active as determined by its cytotoxic effect on murine L-929 fibroblast cells and the bioactivity was retained for at least 6 months in the lysates stored frozen at -20 degrees C in the presence of protease inhibitor PMSF. TNF expressed in P. pastoris was recognized by monoclonal antibodies prepared against recombinant Escherichia coli derived TNF. TNF purified from P. pastoris has the expected N-terminal amino acid sequence and specific activity of 10(7) units/mg protein. TNF analogs were also expressed at levels comparable to that of native TNF. Three of the four analogs were insoluble when produced in P. pastoris.
Similar articles
- High-level expression, purification, and characterization of recombinant human tumor necrosis factor synthesized in the methylotrophic yeast Pichia pastoris.
Sreekrishna K, Nelles L, Potenz R, Cruze J, Mazzaferro P, Fish W, Fuke M, Holden K, Phelps D, Wood P, et al. Sreekrishna K, et al. Biochemistry. 1989 May 2;28(9):4117-25. doi: 10.1021/bi00435a074. Biochemistry. 1989. PMID: 2752013 - Rat tumour necrosis factor-alpha: expression in recombinant Pichia pastoris, purification, characterization and development of a novel ELISA.
Rees GS, Gee CK, Ward HL, Ball C, Tarrant GM, Poole S, Bristow AF. Rees GS, et al. Eur Cytokine Netw. 1999 Sep;10(3):383-92. Eur Cytokine Netw. 1999. PMID: 10477395 - Production and purification of recombinant hirudin expressed in the methylotrophic yeast Pichia pastoris.
Rosenfeld SA, Nadeau D, Tirado J, Hollis GF, Knabb RM, Jia S. Rosenfeld SA, et al. Protein Expr Purif. 1996 Dec;8(4):476-82. doi: 10.1006/prep.1996.0127. Protein Expr Purif. 1996. PMID: 8954896 - The methylotrophic yeast Pichia pastoris as a host for the expression and production of thermostable xylanase from the bacterium Rhodothermus marinus.
Ramchuran SO, Mateus B, Holst O, Karlsson EN. Ramchuran SO, et al. FEMS Yeast Res. 2005 Jun;5(9):839-50. doi: 10.1016/j.femsyr.2004.12.011. Epub 2005 Mar 3. FEMS Yeast Res. 2005. PMID: 15925312 - Pichia pastoris as a host system for transformations.
Cregg JM, Barringer KJ, Hessler AY, Madden KR. Cregg JM, et al. Mol Cell Biol. 1985 Dec;5(12):3376-85. doi: 10.1128/mcb.5.12.3376-3385.1985. Mol Cell Biol. 1985. PMID: 3915774 Free PMC article.
Cited by
- Biochemical and Molecular Characterization of Pichia pastoris Cells Expressing Multiple TMOF Genes (_tmf_A) for Mosquito Larval Control.
Borovsky D, Nauwelaers S, Shatters R Jr. Borovsky D, et al. Front Physiol. 2020 May 26;11:527. doi: 10.3389/fphys.2020.00527. eCollection 2020. Front Physiol. 2020. PMID: 32528316 Free PMC article. - Expression of enzymatically active, recombinant barley alpha-glucosidase in yeast and immunological detection of alpha-glucosidase from seed tissue.
Tibbot BK, Henson CA, Skadsen RW. Tibbot BK, et al. Plant Mol Biol. 1998 Oct;38(3):379-91. doi: 10.1023/a:1006006203372. Plant Mol Biol. 1998. PMID: 9747846 - Overexpression of active Syrian golden hamster prion protein PrPc as a glutathione S-transferase fusion in heterologous systems.
Weiss S, Famulok M, Edenhofer F, Wang YH, Jones IM, Groschup M, Winnacker EL. Weiss S, et al. J Virol. 1995 Aug;69(8):4776-83. doi: 10.1128/JVI.69.8.4776-4783.1995. J Virol. 1995. PMID: 7609044 Free PMC article. - A high-capacity RNA affinity column for the purification of human IRP1 and IRP2 overexpressed in Pichia pastoris.
Allerson CR, Martinez A, Yikilmaz E, Rouault TA. Allerson CR, et al. RNA. 2003 Mar;9(3):364-74. doi: 10.1261/rna.2143303. RNA. 2003. PMID: 12592010 Free PMC article. - Isolation of a novel lipase gene from Serratia liquefaciens S33 DB-1, functional expression in Pichia pastoris and its properties.
Yao H, Yu S, Zhang L, Zuo K, Ling H, Zhang F, Tang K. Yao H, et al. Mol Biotechnol. 2008 Feb;38(2):99-107. doi: 10.1007/s12033-007-9007-6. Epub 2007 Oct 10. Mol Biotechnol. 2008. PMID: 18219590
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources