title: Efficient production from Aspergillus niger of a heterologous protein and an individual protein domain, heavy isotope-labelled, for structure-function analysis.
abstract: 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.
authors.affiliation: Institute of Food Research, Norwich Research Park, Colney, UK.
authors.forename: D A
authors.initials: DA
authors.lastname: MacKenzie
authors.forename: J A
authors.initials: JA
authors.lastname: Spencer
authors.forename: M F
authors.initials: MF
authors.lastname: Le Gal-Coëffet
authors.forename: D B
authors.initials: DB
authors.lastname: Archer
mesh_terms: 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
chemical_list: D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011994:Recombinant Proteins; D009113:Muramidase; D005087:Glucan 1,4-alpha-Glucosidase