Molecular and functional analyses of the metC gene of Lactococcus lactis, encoding cystathionine beta-lyase.
The enzymatic degradation of amino acids in cheese is believed to generate aroma compounds and therefore to be essential for flavor development. Cystathionine beta-lyase (CBL) can convert cystathionine to homocysteine but is also able to catalyze an alpha, gamma elimination. With methionine as a substrate, it produces volatile sulfur compounds which are important for flavor formation in Gouda cheese. The metC gene, which encodes CBL, was cloned from the Lactococcus lactis model strain MG1363 and from strain B78, isolated from a cheese starter culture and known to have a high capacity to produce volatile compounds. The metC gene was found to be cotranscribed with a downstream cysK gene, which encodes a putative cysteine synthase. The MetC proteins of both strains were overproduced in strain MG1363 with the NICE (nisin-controlled expression) system, resulting in a >25-fold increase in cystathionine lyase activity. A disruption of the metC gene was achieved in strain MG1363. Determination of enzymatic activities in the overproducing and knockout strains revealed that MetC is essential for the degradation of cystathionine but that at least one lyase other than CBL contributes to methionine degradation via alpha, gamma elimination to form volatile aroma compounds.
Fernández
M
M
Microbial Ingredients Section, NIZO food research, Ede, The Netherlands.
van Doesburg
W
W
Rutten
G A
GA
Marugg
J D
JD
Alting
A C
AC
van Kranenburg
R
R
Kuipers
O P
OP
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002611:Cheese / Q000382:microbiology; D003001:Cloning, Molecular; D017353:Gene Deletion; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics*; D008190:Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009876:Operon; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D008190:Lyases; C019965:cystathionine beta-lyase