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Applied and Environmental Microbiology, April 2009, p. 2326-2332, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.02417-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Heterologous Production of Methionine-{gamma}-Lyase from Brevibacterium linens in Lactococcus lactis and Formation of Volatile Sulfur Compounds{triangledown} ,{dagger}

Sean B. Hanniffy,1 Mark Philo,1 Carmen Peláez,2 Michael J. Gasson,1 Teresa Requena,2 and M. C. Martínez-Cuesta2*

Food Safety Science Division, Institute of Food Research, Norwich Research Park, Colney Lane, Norwich, Norfolk NR4 7UA, United Kingdom,1 Department of Dairy Science and Technology, Instituto del Frío (CSIC), José Antonio Novais 10, 28420 Madrid, Spain2

Received 21 October 2008/ Accepted 16 February 2009

The conversion of methionine to volatile sulfur compounds (VSCs) is of great importance in flavor formation during cheese ripening and is the focus of biotechnological approaches toward flavor improvement. A synthetic mgl gene encoding methionine-{gamma}-lyase (MGL) from Brevibacterium linens BL2 was cloned into a Lactococcus lactis expression plasmid under the control of the nisin-inducible promoter PnisA. When expressed in L. lactis and purified as a recombinant protein, MGL was shown to degrade L-methionine as well as other sulfur-containing compounds such as L-cysteine, L-cystathionine, and L-cystine. Overproduction of MGL in recombinant L. lactis also resulted in an increase in the degradation of these compounds compared to the wild-type strain. Importantly, gas chromatography-mass spectrometry analysis identified considerably higher formation of methanethiol (and its oxidized derivatives dimethyl disulfide and dimethyl trisulfide) in reactions containing either purified protein, whole cells, or cell extracts from the heterologous L. lactis strain. This is the first report of production of MGL from B. linens in L. lactis. Given their significance in cheese flavor development, the use of lactic acid bacteria with enhanced VSC-producing abilities could be an efficient way to enhance cheese flavor development.

* Corresponding author. Mailing address: Dept. of Dairy Science and Technology, Instituto del Frío, C/José Antonio Novais 10, Ciudad Universitaria, 28040 Madrid, Spain. Phone: 34915492300. Fax: 34915493627. E-mail: cmc{at}if.csic.es

{triangledown} Published ahead of print on 27 February 2009.

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

Applied and Environmental Microbiology, April 2009, p. 2326-2332, Vol. 75, No. 8
0099-2240/09/$08.00+0     doi:10.1128/AEM.02417-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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