Hen egg white lysozyme expressed in, and secreted from, Aspergillus niger is correctly processed and folded

We transformed Aspergillus niger with the full length cDNA gene encoding hen eggwhite lysozyme (HEWL) and its secretion signal sequence. Lysozyme levels up to 12 mg/l were secreted when expression was controlled by the A. awamori glucoamylase (GAM) promoter and 1 mg/l when controlled by the A. nidul...

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Bibliographic Details
Published in:Bio/Technology 1990-08, Vol.8 (8), p.741-745
Main Authors: Archer, D.B. (Agricultural and Food Research Council Institute of Food Research, Norwich, U.K.), Jeenes, D.J, MacKenzie, D.A, Brightwell, G, Lambert, N, Lowe, G, Radford, S.E, Dobson, C.M
Format: Article
Language:English
Subjects:
Agriculture
ALBUMEN D'OEUF
ALBUMINA DE HUEVO
Amino Acid Sequence
Animals
ASPERGILLUS
Aspergillus niger - enzymology
Aspergillus niger - genetics
Bioinformatics
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Chromatography, High Pressure Liquid
Cloning, Molecular - methods
EGG ALBUMEN
Electrophoresis, Polyacrylamide Gel
GENE
Gene Expression
GENES
GENETIC TRANSFORMATION
Glucan 1,4-alpha-Glucosidase - metabolism
Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism
Life Sciences
LISOZIMA
LYSOZYME
Magnetic Resonance Spectroscopy
Molecular Sequence Data
Muramidase - biosynthesis
Muramidase - genetics
Muramidase - secretion
Ovum - enzymology
Plasmids
Protein Conformation
Protein Processing, Post-Translational
PROTEINAS
PROTEINE
PROTEINS
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - secretion
research-paper
Transfection - genetics
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
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Description
Summary:We transformed Aspergillus niger with the full length cDNA gene encoding hen eggwhite lysozyme (HEWL) and its secretion signal sequence. Lysozyme levels up to 12 mg/l were secreted when expression was controlled by the A. awamori glucoamylase (GAM) promoter and 1 mg/l when controlled by the A. nidulans glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter. N-terminal sequence analysis of the recombinant protein indicated that the signal peptide was correctly processed by the A. niger secretory apparatus. The specific catalytic activity of the recombinant protein was identical to that of authentic hen lysozyme. The recombinant HEWL was examined by 2D 1 H-NMR spectroscopy and shown to have a spectrum identical to that of authentic HEWL indicating that the protein was correctly folded.
ISSN:0733-222X
1087-0156
2331-3684
1546-1696
DOI:10.1038/nbt0890-741