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Applied and Environmental Microbiology, July 1999, p. 3251-3257, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Determination of Total Protein Content of Bacterial Cells by SYPRO Staining and Flow Cytometry

Mikhail V. Zubkov,1,2,* Bernhard M. Fuchs,2 Heike Eilers,2 Peter H. Burkill,1 and Rudolf Amann2

Plymouth Marine Laboratory, Plymouth PL1 3DH, United Kingdom,1 and Max-Planck-Institut für Marine Mikrobiologie, D-28359 Bremen, Germany2

Received 22 December 1998/Accepted 20 April 1999

An assay has been developed for measuring protein biomass of marine planktonic bacteria by flow cytometry. The method was calibrated by using five species of Bacteria (an Arcobacter sp., a Cytophaga sp., an Oceanospirillum sp., a Pseudoalteromonas sp., and a Vibrio sp.) recently isolated from seawater samples and grown in culture at different temperatures. The intensity of SYPRO-protein fluorescence of these bacteria strongly correlated with their total protein content, measured by the bicinchoninic acid method to be in the range of 60 to 330 fg of protein cell-1 (r2 = 0.93, n = 34). According to the calibration, the mean biomass of planktonic bacteria from the North Sea in August 1998 was 24 fg of protein cell-1.

* Corresponding author. Mailing address: Plymouth Marine Laboratory, Prospect Place, West Hoe, Plymouth PL1 3DH, United Kingdom. Phone: 44 1752 633422. Fax: 44 1752 633101. E-mail: mvz{at}wpo.nerc.ac.uk.

Applied and Environmental Microbiology, July 1999, p. 3251-3257, Vol. 65, No. 7
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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