Three Diverse Alcohol Dehydrogenases Remain Active at Salt Concentrations Greater than 1 M

Mehran Miroliaei^1, , Rasoul Sharifi² and Peter J. Halling³

¹Developmental and Molecular Biology Division, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran

²Department of Biology, Research and Science Branch, Islamic Azad University, Tehran, Iran

³WestCHEM, Department of Pure & Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, UK

Cite this paper:
Mehran Miroliaei, Rasoul Sharifi and Peter J. Halling. Three Diverse Alcohol Dehydrogenases Remain Active at Salt Concentrations Greater than 1 M. Biomedicine and Biotechnology. 2014; 2(2):37-41. doi: 10.12691/bb-2-2-2

Abstract

A comparative study was carried out on the effects of a number of salts on enzyme activity of three representative alcohol dehydrogenases from non-halophilic sources. The enzymes from yeast (YADH), horse liver (HLADH) and Thermoanaerobacter brockii (TBADH) all retain significant activity at concentrations up to 4 M NaCl. In general catalytic activity follows the order NaCl > Na-acetate > Na₂SO₄ > NaNO₃ > NaClO₄. The deviation from the normal Hofmeister series may reflect effects in line with the “law of matching water affinity”, based on anion interactions with the Na⁺ cation in solution or the essential Zn²⁺ in the enzymes. Retention of activity generally follows the order YADH > HLADH > TBADH, which is opposite to the order of thermostability. Protein structural features promoting thermostability, like additional salt bridges, may lead to greater salt sensitivity. Comparison of cations Cs⁺, K⁺, NH⁴⁺ and Na⁺ showed weaker effects, not clearly in line with the Hofmeister series.

Keywords:
alcohol dehydrogenase hofmeister series thermophilic enzyme salts non-halophilic protein

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References:

[1]	Okamoto, D.N., Kondo, M.Y., Santos, J.A.N., Nakajima, S., Hiraga, K., Oda, K., Juliano, L. Juliano, M.A., and Gouvea, I. E., "Kinetic analysis of salting activation of a subtilisin-like halophilic protease". Biochim. Biophys. Acta, 1794, pp 367-373 (2009).
[2]	L. Rao, X. Zhao, F. Pan, Y. Li, Y. Xue, Y. Ma, J.R. Lu, "Solution Behavior and Activity of a Halophilic Esterase under High Salt Concentration". PLoS One, 14, pp e6 980 (2009).
[3]	N. Coquelle, R. Talon, D. H. Juers, E. Girard, R. Kahn, D. Madern, "Gradual Adaptive Changes of a Protein Facing High Salt Concentrations". J. Mol. Biol., 404, pp 493-505 (2010).
[4]	T. Zhang, S. Datta, J. Eichler, N. Ivanova, S.D. Axen, C.A. Kerfeld, F. Chen, N. Kyrpides, P. Hugenholtz, J.F. Cheng, K.L. Sale, B. Simmons, E. Rubin, "Identification of a haloalkaliphilic and thermostable cellulase with improved ionic liquid tolerance". Green Chem., 13, pp 2083-2090 (2011).
[5]	A. Pennati, G. Zanetti, A. Alivertia, G. Gadda, "Effect of salt and pH on the reductive half-reaction of Mycobacterium tuberculosis FprA with NADPH". Biochemistry, 47, pp 3418-3425 (2008).
[6]	E.M.O. Bowers, L. O. Ragland, L. D. Byers, "Salt effects on beta-glucosidase: pH-profile narrowing". Biochim. Biophys. Acta, 1774, pp 1500-1507 (2007).
[7]	Z. Yang, X.J Liu, C. Chen, P.J. Halling, "Hofmeister effects on activity and stability of alkaline phosphatise". Biochim. Biophys. Acta, 1804, pp 821-828 (2010).
[8]	C.N. Liang, Y. F. Xue, M. Fioroni, F. Rodriguez-Ropero, C. Zhou, U. Schwaneberg, Y. H. Ma, "Cloning and characterization of a thermostable and halo-tolerant endoglucanase from Thermoanaerobacter tengcongensis MB4". Appl. Microbiol. Biotechnol., 89, pp 315-326 (2011).
[9]	N. Eisele, D. Linke, M. Nimtz, R. G. Berger, "Heterologous expression, refolding and characterization of a salt activated subtilase from Pleurotus ostreatus". Process Biochem., 46, pp 1840-1846 (2011).
[10]	A. Basso, C. Cantone, C. Ebert, P.J. Halling, L. Gardossi, "Biocatalysis with undissolved solid substrates and products in organic synthesis with enzymes in non-aqueous media". (2008). G. Carrea and S. Riva, Wiley.
[11]	H. Oneda, Y. Muta, K. Inouye, "Substrate-dependent activation of thermolysin by salt". Biosci. Biotechnol. Biochem., 68, pp 1811-1813 (2004).
[12]	I.A.M. Ahmed, E. E. Babiker, N. Mori. "pH stability and influence of salts on activity of a milk-clotting enzyme from Solanum dubium seeds and its enzymatic action on bovine caseins". LWT-Food Science Technol., 43, pp 759-764 (2010).
[13]	A. Salis, D. Bilanicova, B.W. Ninham, M. Monduzzi, "Hofmeister Effects in Enzymatic Activity: Weak and Strong Electrolyte Influences on the Activity of Candida rugosa Lipase". J. Phys. Chem., 111, pp 1149-1156 (2007).
[14]	D. Bilanicova, A. Salis, B. W. Ninham, M. Monduzzi, "Specific anion effects on enzymatic activity in nonaqueous media". J. Phys. Chem., 112, pp 12066-12072 (2008).
[15]	Y. Zhang, P. S. Cremer, "Interactions between macromolecules and ions: the Hofmeister series". Curr. Opin. Chem. Biol., 10, pp 658-663 (2006).
[16]	K. Nakamura, T, Matsuda, "Biocatalytic reduction of carbonyl groups". Curr. Org. Chem., 10, pp 1217-46 (2006).
[17]	M. Miroliaei, M. Nemat-Gorgani, "Effect of organic solvents on stability and activity of two related alcohol dehydrogenases: a comparative study". Int. J. Biochem. Cell Biol., 34, pp 169-175 (2002).
[18]	K. Ikegaya, "Kinetic analysis about the effects of neutral salts on the thermal stability of yeast alcohol dehydrogenase". J. Biochem., 137, pp 349-354 (2005).
[19]	Y. Cao, L. Liao, X. W. Xu, A. Oren, C. Wang, X. F. Zhu, M. Wu, "Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis". Extremophiles, 12, pp 471-476 (2008).
[20]	T.N. Stekhanova, A.V. Mardanov, E.Y. Bezsudnova, V.M. Gumerov, N.V. Ravin, K.G. Skryabin, V.O. Popov, "Characterization of a Thermostable Short-Chain Alcohol Dehydrogenase from the Hyperthermophilic Archaeon Thermococcus sibiricus". Appl. Environ. Microbiol., 76, pp 4096-4098 (2010).
[21]	M. Negoro, I. Wakabayashi, "Enhancement of alcohol dehydrogenase activity in vitro by acetylsalicylic acid". Eur. J. Pharmacol., 523, pp 25-28 (2005).
[22]	K. Tóth, E. Sedlák, M. Sprinzl, G. Žoldák, "Flexibility and enzyme activity of NADH oxidase from Thermus thermophilus in the presence of monovalent cations of Hofmeister series", Biochim. Biophys. Acta, 1784, pp 789-795 (2008).
[23]	M. Nagaoka, T. Shiraishi, F. Furuhata, Y. Uda, "Effects of inorganic anions on the activation of acid sialidases", Biol. Pharm. Bull., 26, pp 295-298 (2003).
[24]	G. Žoldák, M. Sprinzl, E. Sedlák, "Modulation of activity of NADH oxidase from Thermus thermophilus through change in flexibility in the enzyme active site induced by Hofmeister series anions", Eur. J. Biochem., 271, pp 48-57 (2004).
[25]	A. Barzegara, A.A. Moosavi-Movahedi, J.Z. Pedersen, M. Miroliaei, "Comparative thermostability of mesophilic and thermophilic alcohol dehydrogenases: Stability-determining roles of proline residues and loop conformations". Enzyme Microb. Technol., 45, pp 73-79 (2009).
[26]	Y. Korkhin, A.J. Kalb (Gilboa), M. Peretz, O. Bogin, Y. Burstein, F. Frolow, "NADP-dependent bacterial alcohol dehydrogenases: Crystal structure, cofactor binding and cofactor specificity of the ADHs of Clostridium beijerinckii and Thermoanaerobacter brockii". J. Mol. Biol., 278, pp 965-979 (1998).
[27]	K. Korkhin, O. Bogin, M. Peretz, A.J. Kalb (Gilboa), Y. Burstein, F. Frolow, "Oligomeric integrity—The structural key to thermal stability in bacterial alcohol dehydrogenases". Protein Sci., 8, pp 1241-1249 (1999).
[28]	O. Bogin, I. Levin, Y. Hacham, S. Tel-or, M. Petetz, F. Frolow, Y. Bursten, "Structural basis for the enhanced thermal stability of alcohol dehydrogenase mutants from the mesophilic bacterium Clostridium beijerinckii: contribution of salt bridging". Protein Sci., 11, pp 2561-2574 (2002).
[29]	H. Jörnvall, H. Hans Eklund, C.I. Brändën, "Subunit Conformation of Yeast Alcohol Dehydrogenase". J. Biol. Chem., 235, pp 8414-8419 (1978).
[30]	H. Kirino M. Aoki, M. Aoshima, Y. Hayashi, M. Ohba, A. Yamagishi, T. Wakagi, T. Oshima, "Hydrophobic interaction at the subunit interface contributes to the thermostability of 3-isopropylmalate dehydrogenase froman extreme thermophile, Thermus thermophilus". Eur. J. Biochem., 220, pp 275-281(1994).
[31]	P. Bhattacharyya, K. Santhoshkumar, K. Sharma, "A peptide sequence-YSGVCHTDLHAWHGDWPLPVK [40-60]- in yeast alcohol dehydrogenase prevents the aggregation of denatured substrate proteins". Biochem. Biophys. Res. Com., 307, pp1-7 (2003).
[32]	X. De Bolle, C. Vinals, J. Fastrez, E. Feytmans, "Bivalent cations stabilize yeast alcohol dehydrogenase I". Biochem. J., 323, pp 409-413 (1997).
[33]	H. Eklund, B. Nordstrom, E. Zeppezauer, G. Soderlund, I. Ohlsson, T. Boiwe, B.O. Soderberg, O. Tapia, C.I. Branden, "Three-dimensional structure of horse liver alcohol dehydrogenase at 2.4 A Ê resolution". J. Mol. Biol., 102, pp 27-59 (1976).
[34]	P. Anderson, J. Kvassman, A. Lindstorm, B. Olden, G. Petterson, "Effect of NADH on the pKa of zinc-bound water in liver alcohol dehydrogenase". Eur. J. Biochem., 133, pp 425-433 (1981).
[35]	B.L. Vallee, R.J.P. Williams, "Metalloenzymes: the entatic nature of their active sites". Proc. Natl. Acad. Sci. USA, 59, pp 498-505(1968).
[36]	O. Bogin, M. Peretz, Y. Burstein, "Thermoanaerobacter brockii alcohol dehydrogenase: characterization of the active site metal and its ligand amino acids". Protein Sci., 6, pp 450-458 (1997).
[37]	K.D. Collins, "Ions from the Hofmeister series and osmolytes: effects on proteins in solution and in the crystallization process", Methods, 34, pp 300-311 (2004).
[38]	Boström, M. Williams, D.R.M. and Ninham, B.W. "Specific ion effects: why the properties of lysozyme in salt solutions follow a Hofmeister series", Biophys. J., 85, pp 686-694 (2003).