Overproduction and single-step purification ofBacillus stearothermophilusperoxidase inEscherichia coli

✍ Scribed by Suvit Loprasert; Itaru Urabe; Hirosuke Okada

• Publisher: Springer
• Year: 1990
• Tongue: English
• Weight: 332 KB
• Volume: 32
• Category: Article
• ISSN: 1432-0614
• DOI: 10.1007/bf00164741

✦ Synopsis

The cloned peroxidase gene from Bacillus stearothermophilus was highly expressed in Escherichia coli. Using the high copy n u m b e r plasmid which is tem- perature-sensitive and its own strong promoter, this thermostable peroxidase was p r o d u c e d at 28% o f the total cell proteins when the cells were grown at 42 ° C. The enzyme could be easily purified from E. coli by heat treatment and single-column Sephadex G-200 chromatography. From a 200 ml culture, 30 mg o f purified enzyme was obtained. The peroxidase p r o d u c e d by E. coli s h o w e d a thermostability, haem type and content identical with those o f the peroxidase p r o d u c e d by B. stearothermophilus.