Open AccessA truncated glucoamylase gene fusion for heterologous protein secretion from Aspergillus niger
Author(s) -
Jeenes David J.,
Marczinke Beate,
MacKenzie Donald A.,
Archer David B.
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06041.x
Subject(s) - aspergillus niger , biochemistry , signal peptide , fusion protein , biology , complementary dna , gene , coding region , peptide sequence , recombinant dna
The secreted yield of hen egg‐white lysozyme (HEWL) from the filamentous fungus Aspergillus niger was increased 10–20‐fold by constructing a novel gene fusion. The cDNA sequence encoding mature HEWL was fused in frame to part of the native A. niger gene encoding glucoamylase ( gla A), separated by a proteolytic cleavage site for in vivo processing. Using this construct, peak secreted HEWL yields of 1 g/l were obtained in A. niger shake flask cultures compared to about 50 mg/l when using an expression cassette lacking any gla A coding sequence. The portion of gla A used in the gene fusion encoded the first 498 amino acids of glucoamylase (G498) and comprised its secretion signal, the catalytic domain and most of the O‐glycosylated linker region which, in the entire glucoamylase molecule, spatially separates and links the catalytic and starch‐binding domains.