Expression of hepatitis B surface antigen in the methylotrophic yeast Pichia pastoris using the GAP promoter

Vassileva, Ana; Chugh, Dipti A.; Swaminathan, Sathyamangalam; Khanna, Navin

doi:10.1016/s0168-1656(01)00254-1

Cited by 132 publications

(104 citation statements)

References 23 publications

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“…Increasing the number of copies of the expression cassette generally has the effect of increasing the amount of protein expressed [25,90,112].…”

Section: Gene Dosagementioning

confidence: 99%

“…Multicopy clones containing up to four copies of the HbsAg gene were found to have a four-fold higher yield of HbsAg than those containing a single copy of the gene. No limitation at the transcriptional level was identified [112]. Thus, for this construct the relationship between copy number and product level was very simple.…”

Section: Gene Dosagementioning

confidence: 99%

“…However, an increase in the expression level of this protein was observed using the AOX1 promoter when the gene copy number was increased to two, but levels of expression fell upon further copy number increases. A number of researchers have had significant success with increasing gene dosage utilizing both the AOX1 and GAP promoters [25,70,112], therefore, before considering increasing the gene copy number as an optimization strategy for recombinant protein production from P. pastoris, the identity of the promoter must be considered in advance.…”

Section: Gene Dosagementioning

confidence: 99%

“…Unfortunately, trial and error experiments are often required to find the optimum secretion signal for a specific protein [21]. In proteins such as the hepatitis B surface antigen, which has multiple transmembrane domains, and is likely to be sequestered in the membrane, the use of the α-factor secretion signal did not significantly improve recovery [112].…”

Section: Post-translational Modification Of Secreted Proteinsmentioning

confidence: 99%

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Heterologous protein production using the Pichia pastoris expression system

Macauley-Patrick

Fazenda

McNeil

et al. 2005

Yeast

1,096

633

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The Pichia pastoris expression system is being used successfully for the production of various recombinant heterologous proteins. Recent developments with respect to the Pichia expression system have had an impact on not only the expression levels that can be achieved, but also the bioactivity of various heterologous proteins. We review here some of these recent developments, as well as strategies for reducing proteolytic degradation of the expressed recombinant protein at cultivation, cellular and protein levels. The problems associated with post-translational modifications performed on recombinant proteins by P. pastoris are discussed, including the effects on bioactivity and function of these proteins, and some engineering strategies for minimizing unwanted glycosylations. We pay particular attention to the importance of optimizing the physicochemical environment for efficient and maximal recombinant protein production in bioreactors and the role of process control in optimizing protein production is reviewed. Finally, future aspects of the use of the P. pastoris expression system are discussed with regard to the production of complex membrane proteins, such as G protein-coupled receptors, and the industrial and clinical importance of these proteins.

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“…Increasing the number of copies of the expression cassette generally has the effect of increasing the amount of protein expressed [25,90,112].…”

Section: Gene Dosagementioning

confidence: 99%

Section: Gene Dosagementioning

confidence: 99%

Section: Gene Dosagementioning

confidence: 99%

Section: Post-translational Modification Of Secreted Proteinsmentioning

confidence: 99%

See 2 more Smart Citations

Heterologous protein production using the Pichia pastoris expression system

Macauley-Patrick

Fazenda

McNeil

et al. 2005

Yeast

1,096

633

View full text Add to dashboard Cite

show abstract

“…P. pastoris is a desirable expression system because it has a highly efficient and inducible GAP (glyceraldehyde-3-phosphate dehydrogenase) promoter for high-level expression of proteins. [9][10][11] Hence P. pastoris was used for the high-level production of recombinant OVA.…”

mentioning

confidence: 99%

Structural Characteristics of Hen Egg Ovalbumin Expressed in YeastPichia pastoris

Ito

Matsudomi

2005

Bioscience, Biotechnology, and Biochemistry

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Expression of Recombinant Genes in the Yeast Pichia pastoris

Cregg

Tolstorukov

Kusari

et al. 2010

CP Essential Lab Tech

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The synthesis of specific recombinant proteins using single‐celled organisms from bacteria to mammalian tissue culture cells has become a major source of biopharmaceutical products for the industry and a source of a wide variety of proteins for academic research. A range of organisms are utilized for this purpose. One of the newest and most promising of these is the yeast Pichia pastoris. This article provides detailed basic protocols for the expression of heterologous genes and the synthesis of recombinant proteins utilizing this yeast. Specifically provided are protocols for the insertion of foreign vector DNAs into the yeast by electroporation, amplification of vector sequences by the post‐translational vector amplification (PTVA) method, and growth and expression of foreign genes in shake flask cultures. Curr. Protoc. Essential Lab. Tech. 4:13.2.1‐13.2.14. © 2010 by John Wiley & Sons, Inc.

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Expression of hepatitis B surface antigen in the methylotrophic yeast Pichia pastoris using the GAP promoter

Cited by 132 publications

References 23 publications

Heterologous protein production using the Pichia pastoris expression system

Heterologous protein production using the Pichia pastoris expression system

Structural Characteristics of Hen Egg Ovalbumin Expressed in YeastPichia pastoris

Expression of Recombinant Genes in the Yeast Pichia pastoris

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