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Applied and Environmental Microbiology, October 2009, p. 6488-6495, Vol. 75, No. 20
0099-2240/09/$08.00+0     doi:10.1128/AEM.01055-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Development and Validation of a Species-Independent Functional Gene Microarray That Targets Lactic Acid Bacteria{triangledown}

Stefan Weckx,1,2* Joke Allemeersch,1 Roel Van der Meulen,2 Gino Vrancken,2 Geert Huys,3 Peter Vandamme,3 Paul Van Hummelen,1 and Luc De Vuyst2

MicroArray Facility (MAF), Flanders Institute for Biotechnology (VIB), Herestraat 49, Box 816, B-3000 Leuven, Belgium,1 Research Group of Industrial Microbiology and Food Biotechnology (IMDO), Faculty of Sciences and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium,2 Laboratory of Microbiology, Department of Biochemistry, Physiology and Microbiology, Ghent University, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium3

Received 8 May 2009/ Accepted 9 August 2009

During the last few years, genome-related information has become available for many microorganisms, including important food-related bacteria. Lactic acid bacteria (LAB) are important industrially in the production of fermented foods such as dairy products, sausages, sourdoughs, and vegetables. Despite their limited metabolic capacity, LAB contribute considerably to important characteristics of fermented foods, such as flavor and texture. In the present study, a species-independent functional gene microarray was developed that targets 406 genes that play key roles in the production of sugar catabolites, bacteriocins, exopolysaccharides, and aromas, in probiotic and biosafety characteristics, and in the stress response. Also, genes linked to negative traits, such as antibiotic resistance and virulence, are represented. As LAB ecosystems contain a variety of species, there was a more global focus on these specific functional properties. Thus, an algorithm was used to design gene-specific oligonucleotides that preferably hybridize with multiple LAB species, thereby allowing controlled cross-hybridization. For proof of concept, the microarray composed of 2,269 30-mer oligonucleotides focused on LAB species that are prevalent in sourdough ecosystems. Validation hybridizations using DNA and RNA from 18 LAB strains, covering 86% of all the oligonucleotides, showed that there were wide ranges in intensity and high reproducibility between microarrays.

* Corresponding author. Mailing address: Research Group of Industrial Microbiology and Food Biotechnology (IMDO), Faculty of Sciences and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium. Phone: 32 2 6293863. Fax: 32 2 6292720. E-mail: stefan.weckx{at}vub.ac.be

{triangledown} Published ahead of print on 14 August 2009.

Applied and Environmental Microbiology, October 2009, p. 6488-6495, Vol. 75, No. 20
0099-2240/09/$08.00+0     doi:10.1128/AEM.01055-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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