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Can hTNF-alpha be successfully produced and secreted in filamentous fungus Aspergillus niger?

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Author: Krasevec, N. · Hondel, C.A.M.J.J. van den · Komel, R.
Type:article
Date:2000
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Pflügers Archiv - European Journal of Physiology, 7 suppl., 439, p. R84-R86
Identifier: 56515
Keywords: Aspergillus niger · Heterologous protein secretion · hTNF-α · hTNF-α analogues · Glucan 1,4 alpha glucosidase · Tumor necrosis factor alpha · Amino acid sequence · Aspergillus niger · Biosynthesis · Gene expression · Gene fusion · Genetics · Human · Metabolism · Secretion · Amino Acid Sequence · Artificial Gene Fusion · Aspergillus niger · Gene Expression · Glucan 1,4-alpha-Glucosidase · Humans · Tumor Necrosis Factor-alpha

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. © Springer Verlag 2000 Chemicals/CAS: Glucan 1,4-alpha-Glucosidase, EC 3.2.1.3; Tumor Necrosis Factor-alpha