The use of highly expressed FTH1 as carrier protein for cytosolic targeting in Hansenula polymorpha.
The iron storage protein ferritin is a member of the non-heme iron protein family. It can store and release iron, therefore it prevents the cell from damage caused by iron-dioxygen reactions as well as it provides iron for biological processing. To study whether the human ferritin heavy chain (FTH1) can be expressed in Hansenula polymorpha, we integrated an expression cassette for FTH1 and analyzed the protein expression. We found very efficient expression of FTH1 and obtained yields up to 1.9 g/L under non-optimized conditions. Based on this result we designed a FTH1-PTH fusion protein to successfully express the parathyroid hormone fragment 1-34 (PTH) for the first time intracellular in H. polymorpha.
ARTES Biotechnology GmbH, Elisabeth-Selbert-Str. 9, Langenfeld, Rheinland, Germany. e.eilert@artes-biotechnology.com
Eva
E
Eilert
Cornelis P
CP
Hollenberg
Michael
M
Piontek
Manfred
M
Suckow
D003001:Cloning, Molecular / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D005285:Fermentation; D005293:Ferritins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007501:Iron / Q000378:metabolism; D010088:Oxidoreductases; D010281:Parathyroid Hormone / Q000235:genetics / Q000378:metabolism; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012995:Solubility
D010281:Parathyroid Hormone; D011993:Recombinant Fusion Proteins; D005293:Ferritins; D007501:Iron; C454461:FTH1 protein, human; D010088:Oxidoreductases