This Article Full Text Full Text (PDF) Supplemental material Other Versions of this Article: AEM.00558-07v1 73/16/5292    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Tominaga, T. Articles by Hatakeyama, Y. Search for Related Content PubMed PubMed Citation Articles by Tominaga, T. Articles by Hatakeyama, Y. Agricola Articles by Tominaga, T. Articles by Hatakeyama, Y.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2007, p. 5292-5299, Vol. 73, No. 16
0099-2240/07/$08.00+0     doi:10.1128/AEM.00558-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development of Innovative Pediocin PA-1 by DNA Shuffling among Class IIa Bacteriocins ^,

Saitama Industrial Technology Center North Institute, 2-133 Suehiro, Kumagaya, Saitama 360-0031,¹ Laboratory of Applied Entomology, Department of Plant Science and Resources, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan²

Received 10 March 2007/ Accepted 19 June 2007

Pediocin PA-1 is a member of the class IIa bacteriocins, which show antimicrobial effects against lactic acid bacteria. To develop an improved version of pediocin PA-1, reciprocal chimeras between pediocin PA-1 and enterocin A, another class IIa bacteriocin, were constructed. Chimera EP, which consisted of the C-terminal half of pediocin PA-1 fused to the N-terminal half of enterocin A, showed increased activity against a strain of Leuconostoc lactis isolated from a sour-spoiled dairy product. To develop an even more effective version of this chimera, a DNA-shuffling library was constructed, wherein four specific regions within the N-terminal half of pediocin PA-1 were shuffled with the corresponding sequences from 10 other class IIa bacteriocins. Activity screening indicated that 63 out of 280 shuffled mutants had antimicrobial activity. A colony overlay activity assay showed that one of the mutants (designated B1) produced a >7.8-mm growth inhibition circle on L. lactis, whereas the parent pediocin PA-1 did not produce any circle. Furthermore, the active shuffled mutants showed increased activity against various species of Lactobacillus, Pediococcus, and Carnobacterium. Sequence analysis revealed that the active mutants had novel N-terminal sequences; in active mutant B1, for example, the parental pediocin PA-1 sequence (KYYGNGVTCGKHSC) was changed to TKYYGNGVSCTKSGC. These new and improved DNA-shuffled bacteriocins could prove useful as food additives for inhibiting sour spoilage of dairy products.

Published ahead of print on 25 June 2007.

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