This review summarizes a collection of lactic acid bacteria that are now undergoing genomic sequencing and analysis. Summaries are presented on twenty different species, with each overview discussing the organisms fundamental and practical significance, environmental habitat, and its role in fermentation, bioprocessing, or probiotics. For those projects where genome sequence data were available by March 2002, summaries include 30 a listing of key statistics and interesting genomic features. These efforts will revolutionize our molecular view of Gram-positive bacteria, as up to 15 genomes from the low GC content lactic acid bacteria are expected to be available in the public domain by the end of 2003. Our collective view of the lactic acid bacteria will be fundamentally changed as we rediscover the relationships and capabilities of these organisms through genomics.

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Summary -While catabolism of amino acids is believed to play an important role in cheese f1avor development, the pathways present in cheese microflora are poorly understood. To determine the pathways of aromatic amino acid catabolism in lactococci and effects of Cheddar cheese ripening conditions on catabolic enzymes and products, eight starter lactococcal strains were screened. Cell-free extracts prepared from these strains were found to contain an oe-ketoglutarate-dependent aminotransferase activity with tryptophan, tyrosine and phenylalanine. Tryptophan, tyrosine and phenylalanine aminotransferase specifie activities (Il mol product formed/mg proteinlmin) ranged from 0.30 to 2.8 10-3 , 0.93 to 7.3 10-3 and 1.5 to 7.2 10-3 , respectively. Metabolites produced from tryptophan by a cellfree extract of Lactococcus lactis S3 were indolepyruvic acid, indoleacetic acid and indole-3-aldehyde. Indoleacetic acid and indole-3-aldehyde can form spontaneously from indolepyruvic acid under the conditions employed. A defined medium was used to determine whether the aminotransferase(s) was expressed and which metabolite(s) accumulate under conditions that simulated those of ripening Cheddar cheese in terms of pH, salt, temperature and carbohydrate starvation. The results indicated that the aminotransferase(s) was expressed and stable under these conditions. The tryptophan metabolites that accumulated were determined to be strain-specific. lactococcus / aminotransferase / aromatic amino acid / catabolism / cheese flavor Résumé -Catabolisme des acides aminés aromatiques des lactocoques. Alors que le catabolisme des acides aminés est perçu comme jouant un rôle important dans le développement de la flaveur du fromage; les voies métaboliques liées à la microflore du fromage ne sont que très partiellement éta-blies. Pour déterminer les voies du catabolisme des acides aminés aromatiques dans les lactocoques, et les effets des conditions d'affinage du Cheddar sur les enzymes cataboliques et sur les produits, huit souches de levains lactocoques on été analysées. Des extraits de cellules préparées à partir de ces souches possédaient une activité aminotransférase oe-ketoglutarate dépendante du tryptophane, de la tyrosine et de la phénylalanine. Les activités aminotransférases spécifiques du tryptophane, de la * Oral communication at the IDF Symposium 'Ripening and Quality of Cheeses',

l-Methionine γ-lyase (EC 4.4.1.11 ) was purified to homogeneity from Brevibacterium linens BL2, a coryneform bacterium which has been used successfully as an adjunct bacterium to improve the flavor of Cheddar cheese. The enzyme catalyzes the α,γ elimination of methionine to produce methanethiol, α-ketobutyrate, and ammonia. It is a pyridoxal phosphate-dependent enzyme, with a native molecular mass of approximately 170 kDa, consisting of four identical subunits of 43 kDa each. The purified enzyme had optimum activity at pH 7.5 and was stable at pHs ranging from 6.0 to 8.0 for 24 h. The pure enzyme had its highest activity at 25°C but was active between 5 and 50°C. Activity was inhibited by carbonyl reagents, completely inactivated by dl-propargylglycine, and unaffected by metal-chelating agents. The pure enzyme had catalytic properties similar to those of l-methionine γ-lyase fromPseudomonas putida. Its Km for the catalysis of methionine was 6.12 mM, and its maximum rate of catalysis was 7.0 μmol min−1 mg−1. The enzyme was active under salt and pH conditions found in ripening Cheddar cheese but susceptible to degradation by intracellular proteases.

Methanethiol has been strongly associated with desirable Cheddar cheese flavor and can be formed from the degradation of methionine (Met) via a number of microbial enzymes. Methionine γ-lyase is thought to play a major role in the catabolism of Met and generation of methanethiol in several species of bacteria. Other enzymes that have been reported to be capable of producing methanethiol from Met in lactic acid bacteria include cystathionine β-lyase and cystathionine γ-lyase. The objective of this study was to determine the production, stability, and activities of the enzymes involved in methanethiol generation in bacteria associated with cheese making. Lactococci and lactobacilli were observed to contain high levels of enzymes that acted primarily on cystathionine. Enzyme activity was dependent on the concentration of sulfur amino acids in the growth medium. Met aminotransferase activity was detected in all of the lactic acid bacteria tested and α-ketoglutarate was used as the amino group acceptor. In Lactococcus lactis subsp. cremorisS2, Met aminotransferase was repressed with increasing concentrations of Met in the growth medium. While no Met aminotransferase activity was detected in Brevibacterium linens BL2, it possessed high levels of l-methionine γ-lyase that was induced by addition of Met to the growth medium. Met demethiolation activity at pH 5.2 with 4% NaCl was not detected in cell extracts but was detected in whole cells. These data suggest that Met degradation in Cheddar cheese will depend on the organism used in production, the amount of enzyme released during aging, and the amount of Met in the matrix.