Characterization of recombinant fusion constructs of human b1,4-galactosyltransferase 1 and the lipase pre-propeptide from Staphylococcus hyicus

0313034 - MBÚ 2009 RIV NL eng J - Journal Article
Sauerzapfe, B. - Namdjou, D-J. - Schumacher, T. - Linden, N. - Křenek, Karel - Křen, Vladimír - Elling, L.
Characterization of recombinant fusion constructs of human b1,4-galactosyltransferase 1 and the lipase pre-propeptide from Staphylococcus hyicus.
[Charakterizace rekombinantního fůzního konstruktu lidské b1,4-galactosyltransferasy 1 and the lipasního pre-propeptidu z Staphylococcus hyicus.]
Journal of Molecular Catalysis B-Enzymatic. Roč. 50, 2-4 (2008), s. 128-140. ISSN 1381-1177
R&D Projects: GA AV ČR IAA400200503; GA MŠMT(CZ) LC06010; GA MŠMT OC D25.001
Grant - others:AV ČR(CZ) Bilateral DAAD-AVCR project PPP-D7-CZ 26/04–05D/03/44448
Institutional research plan: CEZ:AV0Z50200510
Keywords : fusion protein * lipase propeptide * staphylococcus hyicus
Subject RIV: CC - Organic Chemistry
Impact factor: 2.015, year: 2008

The question whether proteins fused to b 1,4galactosyltransferase (b4GalT-1) influence the biocatalytic properties of the glycosyltransferase has not been addressed so far. In the present study we have chosen a novel approach to express the gene encoding for human b4GalT-1 from placenta in an N-terminal fusion with the pre-propeptide of the lipase from Staphylococcus hyicus. The pre-propeptide provides a secretion signal in Escherichia coli and was reported to protect fused proteins against proteolytic degradation. Expression of the fusion protein was challenged with an almost full-length version of human b4GalT-1 including parts of the signal anchor and the stem region (propeptide-natb4GalT-1) and the full catalytic domain (His6propeptide-catb4GalT-1), respectively. We demonstrate that the fusion protein in propeptide-natb4GalT-1 is cleaved off during purification using immobilized metal ion chromatography IMAC, most probably catalyzed by the immobilized Zn2+ ions

V této studii je prezentován nový přístup k expresy genu, kódujícímu lidský b4GalT-1 z placenty v N terminalní fůzy s pre-propetidem lipásy z Staphylococcus hyicus
Permanent Link: http://hdl.handle.net/11104/0163959