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Applied and Environmental Microbiology, March 2007, p. 1544-1552, Vol. 73, No. 5
0099-2240/07/$08.00+0     doi:10.1128/AEM.01870-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Expression of Plant Flavor Genes in Lactococcus lactis{triangledown} ,{dagger}

Igor Hernández,1 Douwe Molenaar,1 Jules Beekwilder,2 Harro Bouwmeester,2 and Johan E. T. van Hylckama Vlieg1*

NIZO food research, P.O. Box 20, 6710 BA Ede, The Netherlands,1 Plant Research International, P.O. Box 16, 6700 AA Wageningen, The Netherlands2

Received 7 August 2006/ Accepted 21 December 2006

Lactic acid bacteria, such as Lactococcus lactis, are attractive hosts for the production of plant-bioactive compounds because of their food grade status, efficient expression, and metabolic engineering tools. Two genes from strawberry (Fragaria x ananassa), encoding an alcohol acyltransferase (SAAT) and a linalool/nerolidol synthase (FaNES), were cloned in L. lactis and actively expressed using the nisin-induced expression system. The specific activity of SAAT could be improved threefold (up to 564 pmol octyl acetate h–1 mg protein–1) by increasing the concentration of tRNA1Arg, which is a rare tRNA molecule in L. lactis. Fermentation tests with GM17 medium and milk with recombinant L. lactis strains expressing SAAT or FaNES resulted in the production of octyl acetate (1.9 µM) and linalool (85 nM) to levels above their odor thresholds in water. The results illustrate the potential of the application of L. lactis as a food grade expression platform for the recombinant production of proteins and bioactive compounds from plants.

* Corresponding author. Mailing address: NIZO food research, P.O. Box 20, 6710 BA Ede, The Netherlands. Phone: 31(0)318659511. Fax: 31(0)318650400. E-mail: Johan.van.Hylckama.Vlieg{at}nizo.nl.

{triangledown} Published ahead of print on 5 January 2007.

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

Applied and Environmental Microbiology, March 2007, p. 1544-1552, Vol. 73, No. 5
0099-2240/07/$08.00+0     doi:10.1128/AEM.01870-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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