This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Shao, J.
Right arrow Articles by Wang, P. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shao, J.
Right arrow Articles by Wang, P. G.
Agricola
Right arrow Articles by Shao, J.
Right arrow Articles by Wang, P. G.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, September 2003, p. 5238-5242, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5238-5242.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Enhanced Production of {alpha}-Galactosyl Epitopes by Metabolically Engineered Pichia pastoris

Jun Shao,1 Takahisa Hayashi,2 and Peng George Wang1*

Department of Chemistry, Wayne State University, Detroit, Michigan 48202,1 Wood Research Institute, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan2

Received 18 March 2003/ Accepted 9 June 2003

A metabolically engineered Pichia pastoris strain was constructed that harbored three heterologous enzymes: an S11E mutated sucrose synthase from Vigna radiata, a truncated UDP-glucose C4 epimerase from Saccharomyces cerevisiae, and a truncated bovine {alpha}-1,3-galactosyltransferase. Each gene has its own methanol-inducible alcohol oxidase 1 promoter and transcription terminator on the chromosomal DNA of P. pastoris strain GS115. The proteins were coexpressed intracellularly under the induction of methanol. After permeabilization, the whole P. pastoris cells were used to synthesize {alpha}-galactosyl ({alpha}-Gal) trisaccharide (Gal{alpha}1,3Galß1,4Glc) with in situ regeneration of UDP-galactose. Up to 28 mM {alpha}-Gal was accumulated in a 200-ml reaction. The Pichia system described here is simple and flexible. This work demonstrates that recombinant P. pastoris is an excellent alternative to Escherichia coli transformants in large-scale synthesis of oligosaccharides.

* Corresponding author. Mailing address: Department of Chemistry, Wayne State University, Detroit, MI 48202. Phone: (313) 993-6759. Fax: (313) 577-9241. E-mail: pwang{at}chem.wayne.edu.

Applied and Environmental Microbiology, September 2003, p. 5238-5242, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5238-5242.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»