Functional characterization of a thermostable methionine adenosyltransferase from Thermus thermophilus HB27

Abstract

MATTt (a thermostable methionine adenosyltransferase from Thermus thermophilus HB27) was overexpressed in Escherchia coli and purified using Ni-NTA affinity column. The enzymatic activity of MATTt was investigated in a temperature range from 30 °C to 90 °C, showing that MATTt exhibited a high enzymatic activity and good thermostability at 80 °C. Circular dichroism spectra reveals that MATTt contains high portion of β-sheet structures contributing to the thermostability of MATTt. The kinetic parameter, K_m is 4.19 mmol/L and 1.2 mmol/L for ATP and methionine, respectively. MATTt exhibits the highest enzymatic activity at pH 8. Cobalt (Co²⁺) and zinc ion (Zn²⁺) enhances remarkably the activity of MATTt compared to the magnesium ion (Mg²⁺). All these results indicated that the thermostable MATTt has great potential for industry applications.

Key words： ion-preference; methionine adenosyltransferase; secondary structure; thermostability; Thermus thermophilus

Acknowledgement

This work was financially supported by the National 973 Basic Research Program of China (2014CB745100), National Natural Science Foundation of China (Grant Nos. 21376024 and 21390202).

Electronic Supplementary Material

ƒSupplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s11705-016-1566-2 and is accessible for authorized users.

References

1 Catoni G L. S-Adenosylmethionine: A new intermediate formed enzymatically from L-methionine and adenosinetriphosphate. Journal of Biological Chemistry, 1953, 204: 403–416

2 Fontecave M, Atta M, Mulliez E. S-adenosylmethionine: Nothing goes to waste. Trends in Biochemical Sciences, 2004, 29(5): 243–249

3 Momparler R L, Bovenzi V. DNA methylation and cancer. Journal of Cellular Physiology, 2000, 183: 145–154

4 Dording C M, Mischoulon D, Shyu I. SAMe and sexual functioning. European Psychiatry, 2012, 27(6): 451–454

5 Friedel H A, Goa K L, Benfield P. S-adenosyl-L-methionine: A review of its pharmacological properties and therapeutic potential in liver dysfunction and affective disorders in relation to its physiological role in cell metabolism. Drugs, 1989, 38: 389–416

6 Lieber C S. S-adenosyl-L-methionine: Its role in the treatment of liver disorders. American Journal of Clinical Nutrition, 2002, 76: 1183S–1187S

7 Lu Z J, Markham G D. Enzymatic properties of S-adenosylmethionine synthetase from the archaeon Methanococcus jannaschii. Journal of Biological Chemistry, 2002, 277: 16624–16631

8 Komoto J, Yamada T, Takata Y. Crystal structure of the S-adenosylmethionine synthetase ternary complex: A novel catalytic mechanism of S-adenosylmethionine synthesis from ATP and Met. Biochemistry, 2004, 4: 1821–1831

9 Porcelli M, Caccilapuoti G, Carteni-farina M, Gambacorta A. S-denosylmethionine synthetase in the thermophilic archaebacterium Sulfolobus solfataricus. Purification and characterization of two isoforms. European Journal of Biochemistry, 1988, 177: 273–280

10 Garrido F, Estrela S, Alves S. Refolding and characterization of methionine adenosyltransferase from Euglena gracilis. Protein Expression and Purification, 2011, 79: 128–136

11 Markham G D, Deparsis J, Gatmaitan J. The sequence of metK, the structural gene for S-adenosylmethionine synthetase in Escherichia coli. Journal of Biological Chemistry, 1984, 259: 14505–14507

12 Yoon G S, Ko K H, Kang H W. Characterization of S-adenosylmethionine synthetase from Streptomyces avermitilis NRRL8165 and its effect on antibiotic production. Enzyme and Microbial Technology, 2006, 39(3): 466–473

13 Slapeta J, Stejskal F, Keithly J S. Characterization of s-adenosylmethionine synthetase in Cryptosporidium parvum (apicomplexa). FEMS Microbiology Letters, 2003, 225(2): 271–277

14 Kamarthapu V, Rao K V, Srinivas P N B S, Reddy G B, Reddy V D. Structural and kinetic properties of Bacillus subtilis s-adenosylmethionine synthetase expressed in Escherichia coli. Biochimica et Biophysica Acta, 2008, 1784(12): 1949–1958

15 Burk M J, Van Dien S. Biotechnology for chemical production: Challenges and opportunities. Trends in Biotechnology, 2015, 34(3): 187–190

16 Oshima T, Imahori K. Description of Thermus thermophilus (Yoshida and Oshima) comb. nov., a nonsporulating thermophilic bacterium from a Japanese thermal spa. International Journal of Systematic Bacteriology, 1974, 24: 102–112

17 Liu Y H, Song J N, Tan T W, Liu L. Production of fumaric acid from l-malic acid by solvent engineering using a recombinant thermostable fumarase from Thermus thermophilus HB8. Applied Biochemistry and Biotechnology, 2015, 175(6): 2823–2831

18 Henne A, Brüggemann H, Raasch C. The genome sequence of the extreme thermophile Thermus thermophilus. Nature Biotechnology, 2004, 22: 547–553

19 Chang Y L, Hsieh C L, Huang Y M. Modified method for determination of sulfur metabolites in plant tissues by stable isotope dilution-based liquid chromatography-electrospray ionization-tandem mass spectrometry. Analytical Biochemistry, 2013, 442: 24–33

20 Carolina P, Miguel A A. K2D2: Estimation of protein secondary structure from circular dichroism spectra. BMC Structural Biology, 2008, 8: 25

21 Whitmore L, Wallace B A. Protein secondary structure analyses from circular dichroism spectroscopy: Methods and reference databases. Biopolymers, 2008, 89: 392–400

22 Thompson J D, Gibson T J, Plewniak F. The CLUSTAL_X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 1997, 25: 4876–4882

23 Page R D. TreeView: An application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences: CABIOS, 1996, 12(4): 357–358

24 Deckert G, Warren P V, Gaasterland T. The complete genome of the hyperthermophilic bacterium Aquifex aeolicus. Nature, 1998, 392: 353–358

25 Graham D E, Bock C L, Schalk-Hihi C, Lu Z J, Markham G D. Identification of a highly diverged class of S-adenosylmethionine synthetases in the archaea. Journal of Biological Chemistry, 2000, 275: 4055–4059

26 Markham G D, Hafner E W, Tabor C W. S-Adenosylmethionine synthetase from Escherichia coli. Journal of Biological Chemistry, 1980, 255: 9082–9092

27 Kotb M, Kredich N M. S-Adenosylmethionine synthetase from human lymphocytes. Purification and characterization. Journal of Biological Chemistry, 1985, 260: 3923–3930

28 Takusagawa F, Kamitori S, Misaki S. Crystal structure of S-adenosylmethionine synthetase. Journal of Biological Chemistry, 1996, 271: 136–147

29 Luo Y, Yuan Z, Luo G. Expression of secreted His-tagged S-adenosylmethionine synthetase in the methylotrophic yeast <?Pub Caret?>Pichia pastoris and its characterization, one-step purification, and immobilization. Biotechnology Progress, 2008, 24: 214–220

30 Markham G D, Pajares M A. Structure-function relationships in methionine adenosyltransferases. Cellular and Molecular Life Sciences, 2009, 66: 636–648