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Applied and Environmental Microbiology, November 2009, p. 6688-6695, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.00943-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Engineering of Bacillus subtilis 168 for Increased Nisin Resistance{triangledown} ,{dagger}

Mette E. Hansen,1 Romilda Wangari,1 Egon B. Hansen,2 Ivan Mijakovic,3 and Peter R. Jensen1*

Center for Systems Microbiology, Department of Systems Biology, Technical University of Denmark, DK-2800 Lyngby, Denmark,1 Danisco Innovation Copenhagen, DK-1001 Copenhagen, Denmark,2 Microbiologie et Génétique Moléculaire, AgroParisTech-INRA-CNRS, F-78850 Thiverval-Grignon, France3

Received 27 April 2009/ Accepted 7 September 2009

Nisin is a natural bacteriocin produced commercially by Lactococcus lactis and widely used in the food industry as a preservative because of its broad host spectrum. Despite the low productivity and troublesome fermentation of L. lactis, no alternative cost-effective host has yet been found. Bacillus subtilis had been suggested as a potential host for the biosynthesis of nisin but was discarded due to its sensitivity to the lethal action of nisin. In this study, we have reevaluated the potential of B. subtilis as a host organism for the heterologous production of nisin. We applied transcriptome and proteome analyses of B. subtilis and identified eight genes upregulated in the presence of nisin. We demonstrated that the overexpression of some of these genes boosts the natural defenses of B. subtilis, which allows it to sustain higher levels of nisin in the medium. We also attempted to overcome the nisin sensitivity of B. subtilis by introducing the nisin resistance genes nisFEG and nisI from L. lactis under the control of a synthetic promoter library.

* Corresponding author. Mailing address: Center for Systems Microbiology, Department of Systems Biology, Technical University of Denmark, Building 301, DK-2800 Lyngby, Denmark. Phone: 45 45252510. Fax: 45 45932809. E-mail: prj{at}bio.dtu.dk

{triangledown} Published ahead of print on 11 September 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, November 2009, p. 6688-6695, Vol. 75, No. 21
0099-2240/09/$08.00+0     doi:10.1128/AEM.00943-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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