Optimization of glucose oxidase production by Aspergillus niger using genetic- and process-engineering techniques.
Wild-type Aspergillus niger NRRL-3 was transformed with multiple copies of the glucose oxidase structural gene (god). The gene was placed under the control of the gpdA promoter of A. nidulans. For more efficient secretion the alpha-amylase signal peptide from A. oryzae was inserted in front of god. Compared to the wild type, the recombinant strain NRRL-3 (GOD3-18) produced up to four times more extracellular glucose oxidase under identical culture conditions. Addition of yeast extract (2 gl-1) to a mineral salts medium containing only glucose as carbon source increased volumetric and specific extracellular glucose oxidase activities by 130% and 50% respectively. With the same medium composition and inoculum size, volumetric and specific extracellular glucose oxidase activities increased more than ten times in bioreactor cultivations compared to shake-flask cultures.
Hellmuth
K
K
GBF Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany.
Pluschkell
S
S
Jung
J K
JK
Ruttkowski
E
E
Rinas
U
U
D001233:Aspergillus nidulans / Q000235:genetics; D001234:Aspergillus niger / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D003470:Culture Media / Q000494:pharmacology*; D004867:Equipment Design; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D005813:Genes, Synthetic; D005949:Glucose Oxidase / Q000096:biosynthesis* / Q000235:genetics; D007218:Industrial Microbiology / Q000295:instrumentation / Q000379:methods*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D003470:Culture Media; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D005949:Glucose Oxidase