This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bruinenberg, P. G. Articles by Limsowtin, G. Search for Related Content PubMed PubMed Citation Articles by Bruinenberg, P. G. Articles by Limsowtin, G. Agricola Articles by Bruinenberg, P. G. Articles by Limsowtin, G.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Feb 1997, 561-566, Vol 63, No. 2
Copyright © 1997, American Society for Microbiology

Purification and Characterization of Cystathionine (gamma)-Lyase from Lactococcus lactis subsp. cremoris SK11: Possible Role in Flavor Compound Formation during Cheese Maturation

PG Bruinenberg, G De Roo and GKY Limsowtin
Australian Starter Culture Research Centre Ltd., Werribee, Victoria 3030, Australia

A cystathionine (gamma)-lyase (EC 4.4.1.1) ((gamma)-CTL) was purified to homogeneity from a crude cell extract of Lactococcus lactis subsp. cremoris SK11 by a procedure including anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration chromatography. The activity of SK11 (gamma)-CTL is pyridoxal-5(prm1)-phosphate dependent, and the enzyme catalyzes the (alpha),(gamma)-elimination reaction of L-cystathionine to produce L-cysteine, (alpha)-ketobutyrate, and ammonia. The native enzyme has a molecular mass of approximately 120 to 200 kDa and apparently consists of at least six identical subunits of 20 kDa. In this respect, the SK11 enzyme clearly differs from other bacterial cystathionine lyases, which are all tetrameric proteins with identical subunits of approximately 40 kDa. In addition, the specific activity of purified SK11 (gamma)-CTL toward L-cystathionine is relatively low compared with those reported for other bacterial cystathionine lyases. The SK11 enzyme shows a broad substrate specificity. In the case of L-methionine, the action of SK11 (gamma)-CTL results in the formation of methanethiol, a volatile sulfur compound known to be required in flavor development in cheddar cheese. The (alpha),(beta)-elimination reaction of L-cysteine is also efficiently catalyzed by the enzyme, resulting in the formation of hydrogen sulfide. Although the conditions are far from optimal, cystathionine (gamma)-lyase is still active under cheddar cheese-ripening conditions, namely, pH 5.0 to 5.4 and 5% (wt/vol) NaCl. The possible role of the enzyme in cheese flavor development is discussed.

This article has been cited by other articles:

• Liu, M., Nauta, A., Francke, C., Siezen, R. J. (2008). Comparative Genomics of Enzymes in Flavor-Forming Pathways from Amino Acids in Lactic Acid Bacteria. Appl. Environ. Microbiol. 74: 4590-4600 [Full Text]  
• Irmler, S., Raboud, S., Beisert, B., Rauhut, D., Berthoud, H. (2008). Cloning and Characterization of Two Lactobacillus casei Genes Encoding a Cystathionine Lyase. Appl. Environ. Microbiol. 74: 99-106 [Abstract] [Full Text]  
• Lee, W.-J., Banavara, D. S., Hughes, J. E., Christiansen, J. K., Steele, J. L., Broadbent, J. R., Rankin, S. A. (2007). Role of Cystathionine {beta}-Lyase in Catabolism of Amino Acids to Sulfur Volatiles by Genetic Variants of Lactobacillus helveticus CNRZ 32. Appl. Environ. Microbiol. 73: 3034-3039 [Abstract] [Full Text]  
• Wolle, D. D., Banavara, D. S., Rankin, S. A. (2006). Short Communication: Empirical and Mechanistic Evidence for the Role of Pyridoxal-5'-Phosphate in the Generation of Methanethiol from Methionine. J DAIRY SCI 89: 4545-4550 [Abstract] [Full Text]  
• Martinez-Cuesta, M. C., Pelaez, C., Eagles, J., Gasson, M. J., Requena, T., Hanniffy, S. B. (2006). YtjE from Lactococcus lactis IL1403 Is a C-S Lyase with {alpha},{gamma}-Elimination Activity toward Methionine.. Appl. Environ. Microbiol. 72: 4878-4884 [Abstract] [Full Text]  
• Amarita, F., Yvon, M., Nardi, M., Chambellon, E., Delettre, J., Bonnarme, P. (2004). Identification and Functional Analysis of the Gene Encoding Methionine-{gamma}-Lyase in Brevibacterium linens. Appl. Environ. Microbiol. 70: 7348-7354 [Abstract] [Full Text]  
• Aubel, D., Germond, J. E., Gilbert, C., Atlan, D. (2002). Isolation of the patC gene encoding the cystathionine {beta}-lyase of Lactobacillus delbrueckii subsp. bulgaricus and molecular analysis of inter-strain variability in enzyme biosynthesis. Microbiology 148: 2029-2036 [Abstract] [Full Text]  
• Maicas, S., Ferrer, S., Pardo, I. (2002). NAD(P)H regeneration is the key for heterolactic fermentation of hexoses in Oenococcus oeni. Microbiology 148: 325-332 [Abstract] [Full Text]  
• Liu, Y.-H., Chung, Y.-C., Xiong, Y. (2001). Purification and Characterization of a Dimethoate-Degrading Enzyme of Aspergillus niger ZHY256, Isolated from Sewage. Appl. Environ. Microbiol. 67: 3746-3749 [Abstract] [Full Text]  
• Yvon, M., Chambellon, E., Bolotin, A., Roudot-Algaron, F. (2000). Characterization and Role of the Branched-Chain Aminotransferase (BcaT) Isolated from Lactococcus lactis subsp. cremoris NCDO 763. Appl. Environ. Microbiol. 66: 571-577 [Abstract] [Full Text]  
• Savijoki, K., Palva, A. (2000). Purification and Molecular Characterization of a Tripeptidase (PepT) from Lactobacillus helveticus. Appl. Environ. Microbiol. 66: 794-800 [Abstract] [Full Text]  
• Fernández, M., van Doesburg, W., Rutten, G. A. M., Marugg, J. D., Alting, A. C., van Kranenburg, R., Kuipers, O. P. (2000). Molecular and Functional Analyses of the metC Gene of Lactococcus lactis, Encoding Cystathionine beta -Lyase. Appl. Environ. Microbiol. 66: 42-48 [Abstract] [Full Text]  
• Berger, C., Khan, J. A., Molimard, P., Martin, N., Spinnler, H. E. (1999). Production of Sulfur Flavors by Ten Strains of Geotrichum candidum. Appl. Environ. Microbiol. 65: 5510-5514 [Abstract] [Full Text]  
• Rijnen, L., Bonneau, S., Yvon, M. (1999). Genetic Characterization of the Major Lactococcal Aromatic Aminotransferase and Its Involvement in Conversion of Amino Acids to Aroma Compounds. Appl. Environ. Microbiol. 65: 4873-4880 [Abstract] [Full Text]  
• Turner, M. S., Woodberry, T., Hafner, L. M., Giffard, P. M. (1999). The bspA Locus of Lactobacillus fermentum BR11 Encodes an L-Cystine Uptake System. J. Bacteriol. 181: 2192-2198 [Abstract] [Full Text]  
• Gao, S., Mooberry, E. S., Steele, J. L. (1998). Use of 13C Nuclear Magnetic Resonance and Gas Chromatography To Examine Methionine Catabolism by Lactococci. Appl. Environ. Microbiol. 64: 4670-4675 [Abstract] [Full Text]  
• Dias, B., Weimer, B. (1998). Conversion of Methionine to Thiols by Lactococci, Lactobacilli, and Brevibacteria. Appl. Environ. Microbiol. 64: 3320-3326 [Abstract] [Full Text]