Design of constructs required for succesful anchorage of human enterokinase into the Golgi apparatus

Authors: Hana Hrutková 1   
1 Prírodovedecká fakulta Univerzity Komenského, Bratislava, Slovensko   
Year: 2019
Section: Biotechnology and Food Technology
Abstract No.: 1831
ISBN: 978-80-972360-4-5

The goal of this study was to create production system based on Candida utilis, which would properly process secretion signals. We designed an expression cassette in which we placed the transmembrane domain (TD) at the C-terminus of human enterokinase for efficient anchoring in the membrane of the Golgi apparatus (GA). In the N-terminus we inserted an invertase signal that was enriched downstream with two cleavage sites for natural proteases Kex2 and Ste13. The secretion signal thus should ensure the correct transfer of human enterokinase into the lumen of GA. Another construct, which is the subject of this study, would serve as detection tool for proper integration of human enterokinase into the GA membrane. As it harbours the mutated α-glukosidase with integrated sequence encoding recognition sequence for enterokinase, it should serve very easily for effective detection of enterokinase anchoring into the membrane of GA.

Keywords: heterologous expression; Candida utilis; enterokinase; secretion; detection

Tento projekt bol pripravený na základe projektu „Príprava biologicky aktívnych látok na báze rekombinantných proteínov“ (BIOREKPROT - ITMS: 26240220048) financovaný zo zdojov ERDF a vďaka projektu APVV-17-0149.
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[3] Simeonov P., Berger-Hoffmann R., Hoffmann R., et al. (2011) Protein Eng, Des and Selec, 24(3), p. 261.