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Applied and Environmental Microbiology, August 2000, p. 3566-3573, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Differentiation of Chitinase-Active and Non-Chitinase-Active Subpopulations of a Marine Bacterium during Chitin Degradation

Ace M. Baty III,1,2 Callie C. Eastburn,1,2 Zhenjun Diwu,3 Somkiet Techkarnjanaruk,2 Amanda E. Goodman,4 and Gill G. Geesey1,2,*

Department of Microbiology1 and Center for Biofilm Engineering,2 Montana State University, Bozeman, Montana 59717, Molecular Probes, Inc., Eugene, Oregon 97402,3 and School of Biological Sciences, The Flinders University of South Australia, Adelaide, South Australia 5001, Australia4

Received 30 December 1999/Accepted 4 May 2000

The ability of marine bacteria to adhere to detrital particulate organic matter and rapidly switch on metabolic genes in an effort to reproduce is an important response for bacterial survival in the pelagic marine environment. The goal of this investigation was to evaluate the relationship between chitinolytic gene expression and extracellular chitinase activity in individual cells of the marine bacterium Pseudoalteromonas sp. strain S91 attached to solid chitin. A green fluorescent protein reporter gene under the control of the chiA promoter was used to evaluate chiA gene expression, and a precipitating enzyme-linked fluorescent probe, ELF-97-N-acetyl-beta -D-glucosaminide, was used to evaluate extracellular chitinase activity among cells in the bacterial population. Evaluation of chiA expression and ELF-97 crystal location at the single-cell level revealed two physiologically distinct subpopulations of S91 on the chitin surface: one that was chitinase active and remained associated with the surface and another that was non-chitinase active and released daughter cells into the bulk aqueous phase. It is hypothesized that the surface-associated, non-chitinase-active population is utilizing chitin degradation products that were released by the adjacent chitinase-active population for cell replication and dissemination into the bulk aqueous phase.

* Corresponding author. Mailing address: Center for Biofilm Engineering, 366 EPS Building, Montana State University, Bozeman, MT 59717. Phone: (406) 994-3820. Fax: (406) 994-6098. E-mail: gill_g{at}erc.montana.edu.

Applied and Environmental Microbiology, August 2000, p. 3566-3573, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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