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- Title
Heterologous hyper-expression of a glucansucrase-type glycosyltransferase gene.
- Authors
Swistowska, Anna Maria; Wittrock, Sabine; Collisi, Wera; Hofer, Bernd
- Abstract
Heterologous expression of the large glucansucrase-type glycosyltransferases genes is still a challenge, and typically yields are poor. Therefore, a number of different Escherichia coli systems for the expression of such a gene, encoding the glycosyltransferase R (GtfR) from Streptococcus oralis, were constructed and evaluated. We thereby obtained a strain producing the highest molar yields described so far for this class of enzymes. Cloning of a 5′-terminally truncated version of the gene in the expression vector pET33b(+) yielded, in dissolved form, about 2 μmol (300 mg) of enzyme per liter of culture of an optical density at 600 nm of four. Problems frequently encountered in the heterologous biosynthesis of this class of enzymes, such as formation of a high fraction of insoluble aggregates and/or proteolytic degradation, were not observed in the described system. The over-produced enzyme, devoid of almost its entire variable region, retained its characteristic activities.
- Subjects
GENE expression; GENETIC regulation; GENETIC engineering; BIOCHEMICAL engineering; ENZYMES; TRANSFERASES; BIOSYNTHESIS; ESCHERICHIA coli; BIOCHEMISTRY
- Publication
Applied Microbiology & Biotechnology, 2008, Vol 79, Issue 2, p255
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-008-1435-0