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Applied and Environmental Microbiology, October 2004, p. 5769-5777, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.5769-5777.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Riboflavin Production in Lactococcus lactis: Potential for In Situ Production of Vitamin-Enriched Foods

Catherine Burgess,1 Mary O'Connell-Motherway,1,2 Wilbert Sybesma,3 Jeroen Hugenholtz,3 and Douwe van Sinderen1,2*

Department of Microbiology,1 Alimentary Pharmabiotic Centre, Biosciences Institute, National University of Ireland Cork, Cork, Ireland,2 Wageningen Centre for Food Sciences, Wageningen, The Netherlands3

Received 5 February 2004/ Accepted 9 June 2004

This study describes the genetic analysis of the riboflavin (vitamin B2) biosynthetic (rib) operon in the lactic acid bacterium Lactococcus lactis subsp. cremoris strain NZ9000. Functional analysis of the genes of the L. lactis rib operon was performed by using complementation studies, as well as by deletion analysis. In addition, gene-specific genetic engineering was used to examine which genes of the rib operon need to be overexpressed in order to effect riboflavin overproduction. Transcriptional regulation of the L. lactis riboflavin biosynthetic process was investigated by using Northern hybridization and primer extension, as well as the analysis of roseoflavin-induced riboflavin-overproducing L. lactis isolates. The latter analysis revealed the presence of both nucleotide replacements and deletions in the regulatory region of the rib operon. The results presented here are an important step toward the development of fermented foods containing increased levels of riboflavin, produced in situ, thus negating the need for vitamin fortification.

* Corresponding author. Mailing address: Alimentary Pharmabiotic Centre, Department of Microbiology and Biosciences Institute, National University of Ireland Cork, Western Rd., Cork, Ireland. Phone: 353214901365. Fax: 353214903101. E-mail: d.vansinderen{at}ucc.ie.

Applied and Environmental Microbiology, October 2004, p. 5769-5777, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.5769-5777.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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