國家衛生研究院 NHRI:Item 3990099045/1147

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Title:	Combined use of GAP and AOX1 promoter to enhance the expression of human granulocyte-macrophage colony-stimulating factor in Pichia pastoris
Authors:	Wu, JM;Lin, JC;Chieng, LL;Lee, CK;Hsu, TA
Contributors:	Division of Biotechnology and Pharmaceutical Research
Abstract:	Glyceraldehydes-3-phosphate dehydrogenase (GAP) promoter provides a Strong constitutive expression of heterologous proteins at a level comparable to that seen with the methanol-induced alcohol oxidase (AOXI) promoter in Pichia pastoris. In the present study, strain GS115/G was created to constitutively express secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF) using GAP-based expression system. With addition of 1% casamino acids, the cultivation of GS115/G resulted in ca. 20% higher cells concentration and ca. 30% higher level of secreted protein concentration in YPD medium, The secreted proteins consisted of an unglycosylated and glycosylated recombinant hGM-CSF with different extent of glycosylation. Instead of using GAP promoter alone in strain GS115/G, the strain GS115/GA that could constitutively and inducibly express recombinant hGM-CSF was generated by transforming host strain GS115/G with AOXI promoter-controlled hGM-CSF expression cassette. In the presence of methanol, the strain GS115/GA expressed recombinant hGM-CSF at a level about two-fold higher than that constitutively expressed by GS115/G. (C) 2003 Elsevier Inc. All rights reserved.
Keywords:	Biotechnology & Applied Microbiology
Date:	2003-09-10
Relation:	Enzyme and Microbial Technology. 2003 Sep;33(4):453-459.
Link to:	http://dx.doi.org/10.1016/S0141-0229(03)00147-9
JIF/Ranking 2023:	http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=0141-0229&DestApp=IC2JCR
Cited Times(WOS):	https://www.webofscience.com/wos/woscc/full-record/WOS:000185325800018
Cited Times(Scopus):	http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0042932426
Appears in Collections:	[John Tsu-An Hsu] Conference Papers/Meeting Abstract

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