Document Detail


Efficient secretion of Bacillus subtilis lipase A in Saccharomyces cerevisiae by translational fusion to the Pir4 cell wall protein.
MedLine Citation:
PMID:  18626643     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Both the secretion and the cell surface display of Bacillus subtilis lipase A (Lip A) in Saccharomyces cerevisiae was investigated using different domains of the cell wall protein Pir4 as translational fusion partners. LipA gene minus its leader peptide was fused inframe in two places of PIR4 to achieve cell wall targeting, or substituting most of the PIR4 sequence, after the signal peptide and the Kex2 processed subunit I of Pir4 to achieve secretion to the growth medium. Expression of the recombinant fusion proteins was investigated in a standard and a glycosylation-deficient strain of S. cerevisiae, grown in selective or rich medium. Fusion proteins intended to be retained at the cell wall were secreted to the growth medium, most likely as result of the degradation of the Pir4 moiety containing the cell wall retention domain, giving low levels of lipase activity. However, the fusion intended for secretion was efficiently secreted in a percentage of close to 90% and remained stable even in rich medium at high cell density cultures, yielding values of over 400 IU of lipase activity per milliliter of cell supernatant. This is, to our knowledge, the first report of the efficient production, as a secreted protein, of lipase A of B. subtilis in baker's yeast.
Authors:
María Mormeneo; Isabel Andrés; Cristina Bofill; Pilar Díaz; Jesús Zueco
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-07-15
Journal Detail:
Title:  Applied microbiology and biotechnology     Volume:  80     ISSN:  1432-0614     ISO Abbreviation:  Appl. Microbiol. Biotechnol.     Publication Date:  2008 Sep 
Date Detail:
Created Date:  2008-08-20     Completed Date:  2008-11-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8406612     Medline TA:  Appl Microbiol Biotechnol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  437-45     Citation Subset:  IM    
Affiliation:
Unidad de Microbiología. Facultad de Farmacia, Burjassot, Valencia, Spain.
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MeSH Terms
Descriptor/Qualifier:
Bacillus subtilis / enzymology*
Bacterial Proteins / genetics,  metabolism*
Cell Wall / genetics,  metabolism*
Lipase / genetics,  metabolism*
Protein Transport
Recombinant Fusion Proteins / genetics,  metabolism
Saccharomyces cerevisiae / genetics,  metabolism
Saccharomyces cerevisiae Proteins / genetics,  metabolism*
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Recombinant Fusion Proteins; 0/Saccharomyces cerevisiae Proteins; 0/ccw5 protein, S cerevisiae; EC 3.1.1.3/Lipase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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