Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Nov;61(11):4037–4042. doi: 10.1128/aem.61.11.4037-4042.1995

Purification and Characterization of Cystathionine (beta)-Lyase from Lactococcus lactis subsp. cremoris B78 and Its Possible Role in Flavor Development in Cheese

A C Alting, W Engels, S van Schalkwijk, F A Exterkate
PMCID: PMC1388602  PMID: 16535166

Abstract

An enzyme that degrades sulfur-containing amino acids was purified from Lactococcus lactis subsp. cremoris B78; this strain was isolated from a mixed-strain, mesophilic starter culture used for the production of Gouda cheese. The enzyme has features of a cystathionine (beta)-lyase (EC 4.4.1.8), a pyridoxal-5(prm1)-phosphate-dependent enzyme involved in the biosynthesis of methionine and catalyzing an (alpha),(beta)-elimination reaction. It is able to catalyze an (alpha),(gamma)-elimination reaction as well, which in the case of methionine, results in the production of methanethiol, a putative precursor of important flavor compounds in cheese. The native enzyme has a molecular mass of approximately 130 to 165 kDa and consists of four identical subunits of 35 to 40 kDa. The enzyme is relatively thermostable and has a pH optimum for activity around 8.0; it is still active under cheese-ripening conditions, viz., pH 5.2 to 5.4 and 4% (wt/vol) NaCl. A possible essential role of the enzyme in flavor development in cheese is suggested.

Full Text

The Full Text of this article is available as a PDF (258.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belfaiza J., Parsot C., Martel A., de la Tour C. B., Margarita D., Cohen G. N., Saint-Girons I. Evolution in biosynthetic pathways: two enzymes catalyzing consecutive steps in methionine biosynthesis originate from a common ancestor and possess a similar regulatory region. Proc Natl Acad Sci U S A. 1986 Feb;83(4):867–871. doi: 10.1073/pnas.83.4.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Burnell J. N., Whatley F. R. Sulphur metabolism in Paracoccus denitrificans. Purification, properties and regulation of serine transacetylase, O-acetylserine sulphydrylase and beta-cystathionase. Biochim Biophys Acta. 1977 Mar 15;481(1):246–265. doi: 10.1016/0005-2744(77)90157-7. [DOI] [PubMed] [Google Scholar]
  4. Chopin A. Organization and regulation of genes for amino acid biosynthesis in lactic acid bacteria. FEMS Microbiol Rev. 1993 Sep;12(1-3):21–37. doi: 10.1111/j.1574-6976.1993.tb00011.x. [DOI] [PubMed] [Google Scholar]
  5. Delorme C., Godon J. J., Ehrlich S. D., Renault P. Gene inactivation in Lactococcus lactis: histidine biosynthesis. J Bacteriol. 1993 Jul;175(14):4391–4399. doi: 10.1128/jb.175.14.4391-4399.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dwivedi C. M., Ragin R. C., Uren J. R. Cloning, purification, and characterization of beta-cystathionase from Escherichia coli. Biochemistry. 1982 Jun 22;21(13):3064–3069. doi: 10.1021/bi00256a005. [DOI] [PubMed] [Google Scholar]
  7. Engels D., Ndoricimpa J., Nahimana S., Gryseels B. Control of Schistosoma mansoni and intestinal helminths: 8-year follow-up of an urban school programme in Bujumbura, Burundi. Acta Trop. 1994 Nov;58(2):127–140. doi: 10.1016/0001-706x(94)90052-3. [DOI] [PubMed] [Google Scholar]
  8. Esaki N., Soda K. L-methionine gamma-lyase from Pseudomonas putida and Aeromonas. Methods Enzymol. 1987;143:459–465. doi: 10.1016/0076-6879(87)43081-4. [DOI] [PubMed] [Google Scholar]
  9. Exterkate F. A. Location of Peptidases Outside and Inside the Membrane of Streptococcus cremoris. Appl Environ Microbiol. 1984 Jan;47(1):177–183. doi: 10.1128/aem.47.1.177-183.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gentry-Weeks C. R., Keith J. M., Thompson J. Toxicity of Bordetella avium beta-cystathionase toward MC3T3-E1 osteogenic cells. J Biol Chem. 1993 Apr 5;268(10):7298–7314. [PubMed] [Google Scholar]
  11. Godon J. J., Delorme C., Bardowski J., Chopin M. C., Ehrlich S. D., Renault P. Gene inactivation in Lactococcus lactis: branched-chain amino acid biosynthesis. J Bacteriol. 1993 Jul;175(14):4383–4390. doi: 10.1128/jb.175.14.4383-4390.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Liu S., Pritchard G. G., Hardman M. J., Pilone G. J. Occurrence of arginine deiminase pathway enzymes in arginine catabolism by wine lactic Acid bacteria. Appl Environ Microbiol. 1995 Jan;61(1):310–316. doi: 10.1128/aem.61.1.310-316.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lubkowski J., Wlodawer A., Ammon H. L., Copeland T. D., Swain A. L. Structural characterization of Pseudomonas 7A glutaminase-asparaginase. Biochemistry. 1994 Aug 30;33(34):10257–10265. doi: 10.1021/bi00200a005. [DOI] [PubMed] [Google Scholar]
  14. Mazelis M., Creveling R. K. Purification and properties of S-alkyl-L-cysteine lyase from seedlings of Acacia farnesiana Willd. Biochem J. 1975 Jun;147(3):485–491. doi: 10.1042/bj1470485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nagasawa T., Kanzaki H., Yamada H. Cystathionine gamma-lyase of Streptomyces phaeochromogenes. The occurrence of cystathionine gamma-lyase in filamentous bacteria and its purification and characterization. J Biol Chem. 1984 Aug 25;259(16):10393–10403. [PubMed] [Google Scholar]
  16. Park Y. M., Stauffer G. V. DNA sequence of the metC gene and its flanking regions from Salmonella typhimurium LT2 and homology with the corresponding sequence of Escherichia coli. Mol Gen Genet. 1989 Mar;216(1):164–169. doi: 10.1007/BF00332246. [DOI] [PubMed] [Google Scholar]
  17. Rollema H. S., Kuipers O. P., Both P., de Vos W. M., Siezen R. J. Improvement of solubility and stability of the antimicrobial peptide nisin by protein engineering. Appl Environ Microbiol. 1995 Aug;61(8):2873–2878. doi: 10.1128/aem.61.8.2873-2878.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Uren J. R. Cystathionine beta-lyase from Escherichia coli. Methods Enzymol. 1987;143:483–486. doi: 10.1016/0076-6879(87)43086-3. [DOI] [PubMed] [Google Scholar]
  19. Uren J. R., Ragin R., Chaykovsky M. Modulation of cysteine metabolism in mice--effects of propargylglycine and L-cyst(e)ine-degrading enzymes. Biochem Pharmacol. 1978;27(24):2807–2814. doi: 10.1016/0006-2952(78)90194-6. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES