Hydrogen/deuterium exchange study of subtilisin carlsberg during prolonged exposure to organic solvents

Biotechnology and Bioengineering

Volumn 102, Issue 4, 2009, Pages 1025-1032

  Fasoli, Ezio ^a   Ferrer, Amaris ^a   Barletta, Gabriel L ^a  

^a UNIVERSITY OF PUERTO RICO   (Puerto Rico)

Author keywords

Biocatalysis in organic solvents; Conformational flexibility; Hydrogen deuterium exchange; Subtilisin carlsberg

Indexed keywords

ACTIVITY LOSS; APPARENT RATE CONSTANTS; BIOCATALYSIS IN ORGANIC SOLVENTS; CHANGING DYNAMICS; CHEMICALLY MODIFIED; CONFORMATIONAL FLEXIBILITY; EXCHANGEABLE PROTONS; EXPERIMENTAL CONDITIONS; H/D EXCHANGES; HYDROGEN/DEUTERIUM EXCHANGE; MOBILE PROTONS; PHYSICO-CHEMICAL PROPERTIES; SERINE PROTEASE; SUBTILISIN CARLSBERG; WATER ACTIVITIES;

ACETONITRILE; AMINO ACIDS; ATOMS; CROWN ETHERS; DEUTERIUM; DYNAMICS; ENANTIOSELECTIVITY; ENZYME KINETICS; ENZYMES; ETHERS; ETHYLENE; ETHYLENE GLYCOL; GENE ENCODING; HYDROGEN; ORGANIC SOLVENTS; PROTONS; RATE CONSTANTS; STRUCTURAL DYNAMICS;

ENZYME ACTIVITY;

ACETONITRILE; DIOXANE; MACROGOL; ORGANIC SOLVENT; SERINE PROTEINASE; SUBTILISIN; SUBTILISIN CARLSBERG; UNCLASSIFIED DRUG; 1,4-DIOXANE; ACETONITRILE DERIVATIVE; DEUTERIUM; DIOXANE DERIVATIVE; ENZYME INHIBITOR; HYDROGEN; SOLVENT;

ARTICLE; CATALYST; CHEMICAL MODIFICATION; DEUTERIUM HYDROGEN EXCHANGE; DEUTERON NUCLEAR MAGNETIC RESONANCE; ENANTIOSELECTIVITY; ENZYME ACTIVITY; FREEZE DRYING; CHEMISTRY; KINETICS; METABOLISM; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; UNITED STATES;

ACETONITRILES; DEUTERIUM; DIOXANES; ENZYME INHIBITORS; HYDROGEN; KINETICS; MAGNETIC RESONANCE SPECTROSCOPY; SOLVENTS; SUBTILISINS; UNITED STATES;

EID: 60549109856     PISSN: 00063592     EISSN: 10970290     Source Type: Journal    
DOI: 10.1002/bit.22147     Document Type: Article

References (38)

1. Affleck R, Haynes CA, Clark DS. 1992a. Solvent dielectric effects on protein dynamics. Proc Natl Acad Sci USA 89(11):5167-5170.
2. Affleck R, Xu ZF, Suzawa V, Focht K, Clark DS, Dordick JS. 1992b. Enzymatic catalysis and dynamics in low-water environments. Proc Natl Acad Sci USA 89(3):1100-1104.
3. Broos J, Visser AJWG, Engbersen JFJ, Verboom W, van Hoek A, Reinhoudt DN. 1995. Flexibilityofenzymes suspended in organic solvents probed by time-resolved fluorescence anisotropy. Evidence that enzyme activity and enantioselectivity are directly related to enzyme flexibility. J Am Chem Soc 117(51):12658-12662.
4. Burke PA, Griffin RG, Klibanov A. 1992. Solid-state NMR assessment of enzyme active centers structure under non-aquous conditions. J Biol Chem 267(28):20057-20064.
5. Castillo B, Bansal V, Ganesan A, Halling P, Secundo F, Ferrer A, Griebenow K, Barletta G. 2006. On the relationship between the activity and structure of PEG-alpha-chymotrypsin conjugates in organic solvents. BMC Biotechnol 6:51.
6. Castillo B, Sola RJ, Ferrer A, Barletta G, Griebenow K. 2008. Effect of PEG modification on subtilisin Carlsberg activity, enantioselectivity, and structural dynamics in 1, 4-dioxane. Biotechnol Bioeng 99(1):9-17.
7. Dempsey CE. 1995. Hydrogen bond stabilities in the isolated alamethicin helix: pH-dependent amide exchange measurements in methanol. J Am Chem Soc 117(28):7526-7536.
8. Desai UR, Klibanov AM. 1995. Assessing the structural integrity ofa lyophilized protein in organic solvents. J Am Chem Soc 117(14):3940-3945.
9. Eisenmesser EZ, Bosco DA, Akke M, Kern D. 2002. Enzyme dynamics during catalysis. Science 295(5559):1520-1523.
10. Fasoli E, Castillo B, Santos A, Silva E, Ferrer A, Rosario E, Griebenow K, Secundo F, Barletta GL. 2006. Activation of subtilisin Carlsberg in organic solvents by methyl-β-cyclodextrin: Lyoprotection versus substrate and product-complex effect. J Mol Catalysis B Enzymatic 42(1-2):20-26.
11. Fitzpatrick PA, Klibanov AM. 1991. How can the solvent affect enzyme enantioselectivity? J Am Chem Soc 113(8):3166-3171.
12. Fitzpatrick PA, Ringe D, Klibanov AM. 1992. Computer-assisted modeling of subtilisin enantioselectivity in organic solvents. Biotechnol Bioeng 40(6):735-742.
13. Gekko K, Obu N, Li J, Lee JC. 2004. A linear correlation between the energetics of allosteric communication and protein flexibility in the Escherichia coli cyclic AMP receptor protein revealed by mutation- induced changes in compressibility and amide hydrogen-deuterium exchange. Biochemistry 43(13):3844-3852.
14. Gonzalez S, Alvira E, Cordero LV, Ferrer A, Montales CI, Barletta G. 2002. High initial activity but low storage stability observed for several preparations of subtilisin Carslberg suspended in organic solvents. Biotechnol Prog 18:1462-1466.
15. Griebenow K, Laureano YD, Santos AM, Clemente IM, Rodriguez L, Vidal MW, Barletta G. 1999. Improved enzyme activity and enantioselectivity in organic solvents by methyl-β-cyclodextrin. J Am Chem Soc 121(36):8157-8163.
16. Habeeb AF. 1966 Determination of free amino groups in proteins by trinitrobenzenesulfonic acid. Anal Biochem 14(3):328-336.
17. Henzler-Wildman K, Kern D. 2007. Dynamic personalities of proteins. Nature 450(7172):964-972.
18. Hutcheon GA, Parker MC, Moore BD. 2000. Measuring enzyme motility in organic media using novel H-D exchange methodology. Biotechnol Bioeng 70(3):262-269.
19. Kay LE. 2005. NMR studies of protein structure and dynamics. J Magn Reson 173(2):193-207.
20. Klibanov AM. 2001. Improving enzymes by using them in organic solvents. Nature 409(6817):241-246.
21. Lindorff-Larsen K, Best RB, Depristo MA, Dobson CM, Vendruscolo M. 2005. Simultaneous determination ofprotein structure and dynamics. Nature 433(7022):128-132.
22. Morgan JA, Clark DS. 2004. Salt-activation ofnon-hydrolase enzymes for use in organic solvents. Biotechnol Bioeng 85:456.
23. Partridge J, Dennison PR, Halling PJ, Moore BD. 1998. Subtilisin mobility in hydrophobic organic media measured by proton NMR relaxation: Hydration is more important than solvent dielectric. Biochim Biophys Acta 1386:79-89.
24. Secundo F, Barletta G, Mazzola G. 2008. Role of methoxypolyethylen(glycol) on the hydration, activity, conformation and dynamic properties of a lipase in a dry film. Biotechnol Bioeng 101(2):252-262.
25. Secundo F, Barletta GL, Dumitriu E, Carrea G. 2007. Can an inactivating agent increase enzyme activity in organic solvent? Effects of 18-crown-6 on lipase activity, enantioselectivity, and conformation. Biotechnol Bioeng 97(1):12-18.
26. Smith EL, DeLange RJ, Evans WH, Landon M, Markland FS. 1968. Subtilisin Carlsberg. V. The complete sequence; comparison with subtilisin BPN'; evolutionary relationships. J Biol Chem 243(9):2184-2191.
27. Soares CM, Teixeira VH, Baptista AM. 2003. Protein structure and dynamics in nonaqueous solvents: Insights from molecular dynamics simulation studies. Biophys J 84(3):1628-1641.
28. Sola RJ, Griebenow K. 2006. Chemical glycosylation: New insights on the interrelation between protein structural mobility, thermodynamic stability, and catalysis. FEBS Lett 580(6):1685-1690.
29. Valivety RH, Halling PJ, Macrae AR. 1992. Rhizomucor Miehei lipase remains highly-active at water activity below 0.001. FEBS Lett 301(3):258-260.
30. Valivety RH, Halling PJ, Peilow AD, Macrae AR. 1994. Relationship between water activity and catalytic activity of lipases in organic media.
31. Effects of supports, loading and enzyme preparation. Eur J Biochem 222(2):461-466.
32. van Unen Dirk-Jan, Engbersen JFJ, Reinhoudt DN. 2001. Why do crown ethers activate enzymes in organic solvents? Biotechnol Bioeng 77(3):248-255.
33. Wales TE, Engen JR. 2006. Hydrogen exchange mass spectrometry for the analysis of protein dynamics. Mass Spectrom Rev 25(1):158-170.
34. Watanabe K, Yoshida T, Ueji S. 2001. Optimum conformational flexibility of subtilisin to maximixe the enantioselectivity for subtilisin-catalysed transesterification in an organic solvent with an addition of dimethyl sulfoxide. Chem Commun 14:1260-1261.
35. Wescott CR, Klibanov AM. 1994. The solvent dependence of enzyme specificity. Biochim Biophys Acta 1206(1):1-9.
36. Wu J, Gorenstein DG. 1993. Structure and dynamics of cytochrome c in nonaqueous solvents by 2D NH-exchange NMR spectroscopy. J Am Chem Soc 115(15):6843-6850.
37. Zaks A, Klibanov AM. 1988. The effect of water on enzyme action in organic media. J Biol Chem 263(17):8017-8021.
38. Zavodszky P, Kardos J, Svingor A, Petsko GA. 1998. Adjustment of conformational flexibility is a key event in the thermal adaptation of proteins. Proc Natl Acad Sci USA 95(13):7406-7411.