Author/Editor		Kraševec, Nada; Milunović, Tatjana; Anžur-Lasnik, Marija; Lukančič, Irena; Komel, Radovan; Gaberc-Porekar, Vladka
Title		Human granulocyte colony stimulating factor (G-CSF) produced in the filamentous fungus Aspergillus niger
Type		članek
Vol. and No.		Letnik 61, št. 4
Publication year		2014
Volume		str. 1-9
ISSN		1318-0207 - Acta chimica Slovenica
Language		eng
Abstract		For the first time, a fungal production system is described for expression and secretion of the medically important hu- man protein G-CSF, in Aspergillus niger . A reliable strategy was chosen with in-frame fusion of G-CSF behind a KEX2 cleavage site downstream of the coding region of the highly secreted homologous glucoamylase. This provided secre- tion levels of 5-10 mg/l culture medium of correctly processed G-CSF, although the majority of the protein (>90%) was biologically inactive. Following denaturation/ concentration and chromatographic separation/ renaturation, the G-CSF proliferation activity increased considerably, and analytical immobilised metal affinity chromatography confirmed the monomeric and correctly folded protein. These data suggest that this human secretory protein secreted into the medium of A. niger was not correctly folded, and that it escaped the endoplasmic reticulum folding control systems. This is com- pared to the folding of G-CSF produced in bacteria and yeast.
Keywords		filamentous fungus Aspergillus niger secretion nitasta gliva Aspergillus niger izločanje