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Applied and Environmental Microbiology, January 2003, p. 252-257, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.252-257.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Large-Scale Cultivation of Acidophilic Hyperthermophiles for Recovery of Secreted Proteins

Penny Worthington, Paul Blum,* Francisco Perez-Pomares, and Tom Elthon

School of Biological Sciences, University of Nebraska—Lincoln, Lincoln, Nebraska

Received 25 July 2002/ Accepted 4 October 2002

An electric water heater was modified for large-scale cultivation of aerobic acidophilic hyperthermophiles to enable recovery of secreted proteins. Critical changes included thermostat replacement, redesign of the temperature control circuit, and removal of the cathodic anticorrosion system. These alterations provided accurate temperature and pH control. The bioreactor was used to cultivate selected strains of the archaeon Sulfolobus solfataricus and other species within this genus. Reformulation of a basal salts medium facilitated preparation of large culture volumes and eliminated sterilization-induced precipitation of medium components. Substrate induction of synthesis of the S. solfataricus-secreted alpha-amylase during growth in a defined medium supported the utility of the bioreactor for studies of physiologically regulated processes. An improved purification strategy was developed by using strong cation-exchange chromatography for recovery of the alpha-amylase and the processing of large sample volumes of acidic culture supernatant. These findings should simplify efforts to study acidophilic hyperthermophilic microbes and their secreted proteins.

* Corresponding author. Mailing address: E234 Beadle Center for Genetics, University of Nebraska, Lincoln, NE 68588-0666. Phone: (402) 472-2769. Fax: (402) 472-8722. E-mail: pblum{at}biocomp.unl.edu.

Applied and Environmental Microbiology, January 2003, p. 252-257, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.252-257.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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