Document Detail


Synthesis and accumulation of cyanophycin in transgenic strains of Saccharomyces cerevisiae.
MedLine Citation:
PMID:  18408064     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cyanophycin [multi-L-arginyl-poly(L-aspartic acid) (CGP)] was, for the first time, produced in yeast. As yeasts are very important production organisms in biotechnology, it was determined if CGP can be produced in two different strains of Saccharomyces cerevisiae. The episomal vector systems pESC (with the galactose-inducible promoter GAL1) and pYEX-BX (with the copper ion-inducible promoter CUP1) were chosen to express the cyanophycin synthetase gene from the cyanobacterium Synechocystis sp. strain PCC 6308 (cphA(6308)) in yeast. Expression experiments with transgenic yeasts revealed that the use of the CUP1 promoter is much more efficient for CGP production than the GAL1 promoter. As observed by electrophoresis of isolated CGP in sodium dodecyl sulfate-polyacrylamide gels, the yeast strains produced two different types of polymer: the water-soluble and the water-insoluble CGP were observed as major and minor forms of the polymer, respectively. A maximum CGP content of 6.9% (wt/wt) was detected in the cells. High-performance liquid chromatography analysis showed that the isolated polymers consisted mainly of the two amino acids aspartic acid and arginine and that, in addition, a minor amount (2 mol%) of lysine was present. Growth of transgenic yeasts in the presence of 15 mM lysine resulted in an incorporation of up to 10 mol% of lysine into CGP. Anti-CGP antibodies generated against CGP isolated from Escherichia coli TOP10 harboring cphA(6308) reacted with insoluble CGP but not with soluble CGP, if applied in Western or dot blots.
Authors:
Anna Steinle; Fred Bernd Oppermann-Sanio; Rudolf Reichelt; Alexander Steinbüchel
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-04-11
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  74     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-05-28     Completed Date:  2008-06-26     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3410-8     Citation Subset:  IM    
Affiliation:
Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität, Münster, Germany.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis
Bacterial Proteins
Chromatography, High Pressure Liquid
Cloning, Molecular
Electrophoresis, Polyacrylamide Gel
Gene Expression
Genetic Vectors
Microscopy, Electron, Transmission
Plant Proteins / biosynthesis*,  chemistry,  genetics,  isolation & purification
Recombinant Proteins / biosynthesis,  genetics,  isolation & purification
Saccharomyces cerevisiae / genetics,  metabolism*,  ultrastructure
Synechocystis / genetics
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Bacterial Proteins; 0/Plant Proteins; 0/Recombinant Proteins; 0/cyanophycin
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