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Applied and Environmental Microbiology, August 2008, p. 4889-4897, Vol. 74, No. 15
0099-2240/08/$08.00+0     doi:10.1128/AEM.00393-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Short Peptide Induces an "Uncultivable" Microorganism To Grow In Vitro {triangledown}

D. Nichols,1,2 K. Lewis,1 J. Orjala,3 S. Mo,3 R. Ortenberg,1,2 P. O'Connor,4 C. Zhao,5 P. Vouros,6 T. Kaeberlein,1,2 and S. S. Epstein1,2*

Department of Biology, Northeastern University, Boston, Massachusetts 02115,1 Marine Science Center, Northeastern University, Nahant, Massachusetts 01908,2 Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 900 South Ashland Ave., Chicago, Illinois 60607,3 Biochemistry Department, Boston University School of Medicine, 670 Albany St., 507, Boston, Massachusetts 02118,4 Dept. 09A1, Bldg. AP20, Core R&D Analytical Chemistry, Abbott Laboratories, 100 Abbott Park Rd., Abbott Park, Illinois 60064,5 Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts 021156

Received 15 February 2008/ Accepted 16 May 2008

Microorganisms comprise the bulk of biodiversity, but only a small fraction of this diversity grows on artificial media. This phenomenon was noticed almost a century ago, repeatedly confirmed, and termed the "great plate count anomaly." Advances in microbial cultivation improved microbial recovery but failed to explain why most microbial species do not grow in vitro. Here we show that at least some of such species can form domesticated variants capable of growth on artificial media. We also present evidence that small signaling molecules, such as short peptides, may be essential factors in initiating growth of nongrowing cells. We identified one 5-amino-acid peptide, LQPEV, that at 3.5 nM induces the otherwise "uncultivable" strain Psychrobacter sp. strain MSC33 to grow on standard media. This demonstrates that the restriction preventing microbial in vitro growth may be different from those offered to date to explain the "great plate count anomaly," such as deficiencies in nutrient composition and concentrations in standard media, medium toxicity, and inappropriate incubation time. Growth induction of MSC33 illustrates that some microorganisms do not grow in vitro because they are removed from their native communities and the signals produced therein. "Uncultivable" species represent the largest source of unexplored biodiversity, and provide remarkable opportunities for both basic and applied research. Access to cultures of some of these species should be possible through identification of the signaling compounds necessary for growth, their addition to standard medium formulations, and eventual domestication.

* Corresponding author. Mailing address: Department of Biology, Northeastern University, 360 Huntington Avenue, Boston, MA 02115. Phone: (617) 373-4048. Fax: (617) 373-3724. E-mail: s.epstein{at}neu.edu

{triangledown} Published ahead of print on 30 May 2008.

Applied and Environmental Microbiology, August 2008, p. 4889-4897, Vol. 74, No. 15
0099-2240/08/$08.00+0     doi:10.1128/AEM.00393-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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