Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
11
pubmed:dateCreated
2006-11-7
pubmed:abstractText
Yeast cell surface display is a powerful tool for expression and immobilization of biocatalytically active proteins on a unicellular eukaryote. Here bacterial carboxylesterase EstA from Burkholderia gladioli was covalently anchored into the cell wall of Saccharomyces cerevisiae by in-frame fusion to the endogenous yeast proteins Kre1p, Cwp2p, and Flo1p. When p-nitrophenyl acetate was used as a substrate, the esterase specific activities of yeast expressing the protein fusions were 103 mU mg(-1) protein for Kre1/EstA/Cwp2p and 72 mU mg(-1) protein for Kre1/EstA/Flo1p. In vivo cell wall targeting was confirmed by esterase solubilization after laminarinase treatment and immunofluorescence microscopy. EstA expression resulted in cell wall-associated esterase activities of 2.72 U mg(-1) protein for Kre1/EstA/Cwp2p and 1.27 U mg(-1) protein for Kre1/EstA/Flo1p. Furthermore, esterase display on the yeast cell surface enabled the cells to effectively grow on the esterase-dependent carbon source glycerol triacetate (Triacetin). In the case of Kre1/EstA/Flo1p, in vivo maturation within the yeast secretory pathway and final incorporation into the wall were further enhanced when there was constitutive activation of the unfolded protein response pathway. Our results demonstrate that esterase cell surface display in yeast, which, as shown here, is remarkably more effective than EstA surface display in Escherichia coli, can be further optimized by activating the protein folding machinery in the eukaryotic secretion pathway.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-11152069, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-11956747, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-12007643, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-12200308, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-12466889, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-12637735, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-12676684, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-12888957, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-14716465, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-2744488, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-6336730, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-8390128, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-8511970, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-8735548, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-8898193, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-8936431, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-9023939, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-9035102, http://linkedlifedata.com/resource/pubmed/commentcorrection/16980424-9684341
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0099-2240
pubmed:author
pubmed:issnType
Print
pubmed:volume
72
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
7140-7
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2006
pubmed:articleTitle
Cell surface expression of bacterial esterase A by Saccharomyces cerevisiae and its enhancement by constitutive activation of the cellular unfolded protein response.
pubmed:affiliation
Angewandte Molekularbiologie, FR 8.3, Gebäude A1 5, Universität des Saarlandes, Postfach 151150, D-66041 Saarbrücken, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Evaluation Studies