Efficient production from Aspergillus niger of a heterologous protein and an individual protein domain, heavy isotope-labelled, for structure-function analysis.
Aspergillus niger has been used successfully to secrete proteins labelled with 13C and/or 15N to a specific activity of > 99% for high resolution NMR analysis. In the case of a heterologous protein, hen egg-white lysozyme, 15N single-labelled and 13C, 15N double-labelled forms were secreted at yields of 100-200 mg l-1 by optimising the type of carbon source used and the ratio of carbon to nitrogen. Another protein, the glucoamylase starch-binding domain from A. niger, was also produced as the 15N single-labelled form at 20-40 mg l-1.
MacKenzie
D A
DA
Institute of Food Research, Norwich Research Park, Colney, UK.
Spencer
J A
JA
Le Gal-Coëffet
M F
MF
Archer
D B
DB
D001234:Aspergillus niger / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002247:Carbon Isotopes; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D007553:Isotope Labeling; D009682:Magnetic Resonance Spectroscopy; D009113:Muramidase / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D009113:Muramidase; D005087:Glucan 1,4-alpha-Glucosidase