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Applied and Environmental Microbiology, March 2003, p. 1457-1464, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1457-1464.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Use of a Packed-Column Bioreactor for Isolation of Diverse Protease-Producing Bacteria from Antarctic Soil

Nathalie Wery,1* Ursula Gerike,1 Ajay Sharman,2 Julian B. Chaudhuri,3 David W. Hough,1 and Michael J. Danson1

Centre for Extremophile Research, Department of Biology and Biochemistry,1 Centre for Extremophile Research, Department of Chemical Engineering, University of Bath, Bath BA2 7AY,3 Reckitt Benckiser Plc., Hull HU7 8DS, United Kingdom2

Received 2 August 2002/ Accepted 19 December 2002

Seventy-five aerobic heterotrophs have been isolated from a packed-column bioreactor inoculated with soil from Antarctica. The column was maintained at 10°C and continuously fed with a casein-containing medium to enrich protease producers. Twenty-eight isolates were selected for further characterization on the basis of morphology and production of clearing zones on skim milk plates. Phenotypic tests indicated that the strains were mainly psychrotrophs and presented a high morphological and metabolical diversity. The extracellular protease activities tested were optimal at neutral pH and between 30 and 45°C. 16S ribosomal DNA sequence analyses showed that the bioreactor was colonized by a wide variety of taxons, belonging to various bacterial divisions: {alpha}-, ß-, and {gamma}-Proteobacteria; the Flexibacter-Cytophaga-Bacteroides group; and high G+C gram-positive bacteria and low G+C gram-positive bacteria. Some strains represent candidates for new species of the genera Chryseobacterium and Massilia. This diversity demonstrates that the bioreactor is an efficient enrichment tool compared to traditional isolation strategies.

* Corresponding author. Mailing address: Centre for Extremophile Research, Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom. Phone: 44-1225-386509. Fax: 44-1225-386779. E-mail: bssmjd{at}bath.ac.uk.

Applied and Environmental Microbiology, March 2003, p. 1457-1464, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1457-1464.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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