Title
Intracellular and extracellular production of proteins in Aspergillus under the control of expression signals of the highly expressed Aspergillus nidulans gpdA gene
Author
Punt, P.J.
Zegers, N.D.
Busscher, M.
Pouwels, P.H.
van den Hondel, C.A.M.J.J.
Medisch Biologisch Laboratorium TNO
Publication year
1991
Abstract
The expression in Aspergillus is described of genes, coding for intracellular and extracellular proteins controlled by the promoter region of the constitutively and efficiently expressed glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) of Aspergillus nidulans. Both the homologous gpdA and the heterologous Escherichia coli β-galactosidase (lacZ) and β-glucuronidase (uidA) genes could be expressed intracellularly at levels as high as 10-25% of total soluble protein. Efficient extracellular production of A. niger glucoamylase could be achieved with a fusion-gene containing the region of the glucoamylase gene coding for the mature protein preceded by a synthetic fungal signal sequence. Extracellular production of a heterologous protein, E. coli β-glucuronidase, with such a fusion was much less efficient. Only very low levels of β-glucuronidase were detected in the culture fluid, whereas considerable enzyme activity was detected in the mycelium. Chemicals/CAS: Bacterial Proteins; beta-Galactosidase, EC 3.2.1.23; Fungal Proteins; Genetic Vectors; Glucan 1,4-alpha-Glucosidase, EC 3.2.1.3; Glucosephosphate Dehydrogenase, EC 1.1.1.49; Glucuronidase, EC 3.2.1.31; Recombinant Fusion Proteins
Subject
E. coli β-galactosidase
E. coli β-glucuronidase
Filamentous fungi
Glucoamylase
Recombinant DNA
Signal sequence Glyceraldehyde-3-phosphate dehydrogenase
Aspergillus nidulans
Gene control
Gene expression
Nonhuman
Priority journal
Protein secretion
Amino Acid Sequence
Aspergillus nidulans
Aspergillus niger
Bacterial Proteins
Base Sequence
Beta-Galactosidase
Comparative Study
Enzyme Induction
Escherichia coli
Fungal Proteins
Gene Expression Regulation, Fungal
Genes, Fungal
Genetic Vectors
Glucan 1,4-alpha-Glucosidase
Glucosephosphate Dehydrogenase
Glucuronidase
Molecular Sequence Data
Promoter Regions (Genetics)
Recombinant Fusion Proteins
Aspergillus
Aspergillus niger
Emericella nidulans
Escherichia coli
Fungi
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DOI
https://doi.org/10.1016/0168-1656(91)90024-p
TNO identifier
231448
ISSN
0168-1656
Source
Journal of Biotechnology, 17 (1), 19-33
Bibliographical note
Correspondence Address: Punt, P.J.; TNO Medical Biological Lab., PO Box 45, 2280 AA Rijswijk, Netherlands
Document type
article