Can hTNF-alpha be successfully produced and secreted in filamentous fungus Aspergillus niger?
A gene-fusion expression strategy was applied for the heterologous expression of hTNF-alpha in A. niger AB1.13. The TNF-alpha 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-alpha 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-alpha 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.
Krasevec
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N
National Institute of Chemistry, Ljubljana, Slovenia.
van den Hondel
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CA
Komel
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D000595:Amino Acid Sequence / Q000235:genetics; D019677:Artificial Gene Fusion; D001234:Aspergillus niger / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics; D006801:Humans; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism
D014409:Tumor Necrosis Factor-alpha; D005087:Glucan 1,4-alpha-Glucosidase