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Published on: March 15, 2012
Can hTNF-α be successfully produced and secreted in filamentous fungus Aspergillus niger?
Nada Kraševec1, Cees A M J J van den Hondel2, Radovan Komel1
1National Institute of Chemistry, Hajdrihova 19, SI-1000 Ljubljana, Slovenia, , , , , , SI.
Abstract:
A gene-fusion expression strategy was applied for the heterologous expression of hTNF-α in A. niger AB1.13. The TNF-α gene was fused with the A. niger glucoamylase GII form as a carrier-gene, behind its transcription control and secretion signal. The protein was expressed in the cells in the form of a glucoamylase-fusion protein, but was not present in the culture medium. From the expression of two hTNF-α analogues, LK 811 (Cys95/148) and LK 802 (Cys95/148, His107/108) respectively, we concluded that oligomerisation was not the critical point for secretion of hTNF-α in A. niger, but more probably improper folding already at the stage of monomer formation, or even incorrect processing of the molecule during the secretion pathway.
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