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Antimicrobial Agents and Chemotherapy, May 2009, p. 1964-1973, Vol. 53, No. 5
0066-4804/09/$08.00+0     doi:10.1128/AAC.01382-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Novel Mechanism for Nisin Resistance via Proteolytic Degradation of Nisin by the Nisin Resistance Protein NSR{triangledown}

Zhizeng Sun,1,2 Jin Zhong,1* Xiaobo Liang,1,{dagger} Jiale Liu,1 Xiuzhu Chen,1 and Liandong Huan1*

State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China,1 Graduate University, Chinese Academy of Sciences, Beijing 100039, China2

Received 15 October 2008/ Returned for modification 8 January 2009/ Accepted 27 February 2009

Nisin is a 34-residue antibacterial peptide produced by Lactococcus lactis that is active against a wide range of gram-positive bacteria. In non-nisin-producing L. lactis, nisin resistance could be conferred by a specific nisin resistance gene (nsr), which encodes a 35-kDa nisin resistance protein (NSR). However, the mechanism underlying NSR-mediated nisin resistance is poorly understood. Here we demonstrated that the protein without the predicted N-terminal signal peptide sequence, i.e., NSRSD, could proteolytically inactivate nisin in vitro by removing six amino acids from the carboxyl "tail" of nisin. The truncated nisin (nisin1-28) displayed a markedly reduced affinity for the cell membrane and showed significantly diminished pore-forming potency in the membrane. A 100-fold reduction of bactericidal activity was detected for nisin1-28 in comparison to that for the intact nisin. In vivo analysis indicated that NSR localized on the cell membrane and endowed host strains with nisin resistance by degrading nisin as NSRSD did in vitro, whereas NSRSD failed to confer resistance upon the host strain. In conclusion, we showed that NSR is a nisin-degrading protease. This NSR-mediated proteolytic cleavage represents a novel mechanism for nisin resistance in non-nisin-producing L. lactis.


* Corresponding author. Mailing address: Institute of Microbiology, Chinese Academy of Sciences, A3 Datun Road, Chaoyang District, Beijing 100101, China. Phone: (86) 10 6480 7439. Fax: (86) 10 6480 7401. E-mail for Jin Zhong: zhongj{at}im.ac.cn. E-mail for Liandong Huan: huanld{at}im.ac.cn

{triangledown} Published ahead of print on 9 March 2009.

{dagger} Present address: Department of Pediatric Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294.


Antimicrobial Agents and Chemotherapy, May 2009, p. 1964-1973, Vol. 53, No. 5
0066-4804/09/$08.00+0     doi:10.1128/AAC.01382-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.