Fermentation of a yeast producing A. niger glucose oxidase: scale-up, purification and characterization of the recombinant enzyme.
We have developed a fermentation process to produce up to 3 grams per liter of active, secreted glucose oxidase from a recombinant Saccharomyces cerevisiae. Real-time size-exclusion HPLC analysis is used to monitor enzyme production during fermentation, and purification to more than 95 percent is obtained using only filtration methods. The recombinant enzyme is stable to higher temperatures and a wider pH range than the native Aspergillus niger enzyme, and is free of contaminating amylase, cellulase and catalase.
De Baetselier
A
A
Chiron Research Labs, Chiron Corporation, Emeryville, CA 94608.
Vasavada
A
A
Dohet
P
P
Ha-Thi
V
V
De Beukelaer
M
M
Erpicum
T
T
De Clerck
L
L
Hanotier
J
J
Rosenberg
S
S
D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics; D001709:Biotechnology; D004355:Drug Stability; D005285:Fermentation; D005949:Glucose Oxidase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D010957:Plasmids; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics
D011994:Recombinant Proteins; D005949:Glucose Oxidase