Abstract
To overcome the extracellular salt stress, Methanohalophilus portucalensis FDF1T synthesizes the compatible solute betaine through the methylation of glycine, sarcosine, and N,N-dimethylglycine. S-adenosylmethionine (AdoMet) is the methyl donor. The enzyme sarcosine dimethylglycine N-methyltransferase (SDMT) of M. portucalensis, that catalyzes the formation of N,N-dimethylglycine and glycine betaine, has been purified and characterized. SDMT, a monomer of 33 kDa with a pI at 5.03, has a narrow substrate specificity limited to using only sarcosine and dimethylglycine as substrates for the methyl transferase reaction. The K m values for sarcosine and AdoMet were 2.29 and 0.21 mM, respectively, with a V max of 0.83 μmol/mg-min (k cat value of 0.44 s−1). The K m values for dimethylglycine and AdoMet were 3.76 and 0.59 mM, respectively, with a V max of 4.88 μmol/mg-min (k cat of 2.68 s−1). A high concentration of the end product betaine (2.0 M) did not affect the SMT activity, but it slightly inhibited the DMT activity. Both activities were also not affected by potassium or sodium ions in concentrations of 200–1,000 mM. We compared this novel archaeal SDMT enzyme to other similar bacterial transferases as well as to the glycine sarcosine dimethylglycine methyltransferase found also in M. portucalensis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Anthoni U, Christophersen C, Horegard L, Nielsen PH (1991) Quaternary ammonium compounds in the biosphere-an example of a versatile adaptive strategy. Comp Biochem Physiol 99B:1–18
Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS (1979) Methanogens: reevaluation of a unique biological group. Microbiol Rev 43:260–296
Boone DR, Mathrani IM, Liu Y, Menaia JAGF, Mah RA, Boone JE (1993) Isolation and characterization of Methanohalophilus portucalensis sp. nov. and DNA reassociation study of the genus Methanohalophilus. Int J Syst Bact 43:430–437
Csonka LN, Hanson AD (1991) Prokaryotic osmoregulation: genetics and physiology. Annu Rev Microbiol 45:569–606
Galinski EA (1993) Compatible solutes of halophilic eubacteria: molecular principles, water-solute interaction, stress protection. Experientia 49:487–496
Javor B (1989) Hypersaline environments: microbiology and biochemistry. Springer, New York, pp 101–124
Kinne RKH (1993) The role of organic osmolytes in osmoregulation: from bacteria to mammals. J Exp Zoo 265:346–355
Lai MC, Gunsalus RP (1992) Glycine betaine and potassium ion are the major compatible solutes in the extremely halophilic methanogen Methanohalophilus strain Z7302. J Bacteriol 174:7474–7477
Lai MC, Sowers KR, Robertson DE, Roberts MF, Gunsalus RP (1991) Distribution of compatible solutes in the halophilic methanogenic archaebacteria. J Bacteriol 173:5352–5358
Lai MC, Yang DR, Chuang MJ (1999) Regulatory factors associated with synthesis of the osmolyte glycine betaine in the halophilic methanoarchaeon Methanohalophilus portucalensis. Appl Environ Microbiol 65:828–833
Lai MC, Hong TY, Gunsalus RP (2000) Glycine betaine transport in the obligate Halophilic Archaeon—Methanohalophilus portucalensis. J Bacteriol 182:5020–5024
Lai MC, Wang CC, Chuang MJ, Wu YC, Lee YC (2006) Effects of substrate and potassium on the betaine-synthesizing enzyme glycine sarcosine dimethylglycine N-methyltransferase from a halophilic methanoarchaeon Methanohalophilus portucalensis. Res Microbiol 67:1–8
Lu WD, Chi ZM, Su CD (2006) Identification of glycine betaine as compatible solute in Synechococcus sp. WH8102 and characterization of its N-methyltransferase genes involved in betaine synthesis. Arch Microbiol 186:495–506
Martin JL, McMillan FM (2002) SAM (dependent) I AM: the S-adenosyl-methionine-dependent methyltransferase fold. Curr Opin Struct Biol 12:783–793
Mathrani IM, Boone DR, Mah MA (1985) Isolation and characterization of a moderately halophilic methanogen from a solar saltern. Appl Environ Microbiol 50:140–143
Nyyssölä A, Leisola M (2001) Actinopolyspora halophila has two separate pathways to betaine synthesis. Arch Microbiol 176:294–300
Nyyssölä A, Kerovuo J, Kaukinen P, von Weymarn N, Reinikaiuem T (2000) Extreme halophiles synthetic betaine from glycine by methylation. J Biol Chem 275:22196–22201
Nyyssölä A, Reinikaiuem T, Leisola M (2001) Characterization of glycine sarcosine N-methyltransferase and sarcosine dimethylglycine N-methyltransferase. Appl Environ Microbiol 67:2044–2050
Ogawa H, Gomi T, Takusagawa F, Fujioka M (1998) Structure, function and physiological role of glycine N-methyltransferase. Int J Biochem Cell Biol 30:13–26
Ollivier B, Canmette P, Garcia JL, Horiuti Y (1994) Anaerobic bacteria from hypersaline environments. Microbiol Rev 58:27–38
Proctor LM, Lai R, Gunsalus RP (1997) The methanogenic archaeon Methanosarcina thermophila TM-1 possesses a high-affinity glycine betaine transporter involved in osmotic adaptation. Appl Environ Microbiol 63:2252–2257
Roberts MF (2000) Osmoadaptation and osmoregulation in archaea. Front Biosci 5:796–812
Roberts MF, Lai MC, Gunsalus RP (1992) Biosynthetic pathway of the osmolytes N ε-acetyl-β-lysine, β-glutamine, and betaine in Methanohalophilus strain FDF1 suggested by nuclear magnetic resonance analyses. J Bacteriol 174:6688–6693
Roeßler M, Müller V (2001) Osmoadaptation in bacteria and archaea: common principles and differences. Environ Microbiol 3:743–754
Roeßler M, Pflüger K, Flach H, Lienard T, Gottschalk G, Müller V (2002) Identification of a salt-induced primary transporter for glycine betaine in the Methanogen Methanosarcina mazei Gö1. Appl Environ Microbiol 68:2133–2139
Sleator RD, Hill C (2001) Bacterial osmoadaptation: the role of osmolytes in bacterial stress and virulence. FEMS Microbiol Rev 26:49–71
Waditee R, Tanaka Y, Aoki K, Hibino T, Jikuya H, Takano J, Takabe T, Takabe T (2003) Isolation and functional characterization of N-methyltransferases that catalyze betaine synthesis from glycine in a halotolerant photosynthetic organism Aphanothece halophytica. J Biol Chem 278:4932–49432
Acknowledgments
This work was supported by National Science Council (Taiwan, ROC) grant 96-2311-B-005-002 to MCL.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by William Metcalf.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chen, SY., Lai, MC., Lai, SJ. et al. Characterization of osmolyte betaine synthesizing sarcosine dimethylglycine N-methyltransferase from Methanohalophilus portucalensis . Arch Microbiol 191, 735–743 (2009). https://doi.org/10.1007/s00203-009-0501-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-009-0501-z