Active site mutants of pyruvate decarboxylase from Zymomonas mobilis - A site-directed mutagenesis study of L112, I472, I476, E473 and N482

European Journal of Biochemistry

Volumn 257, Issue 3, 1998, Pages 538-546

  Pohl, Martina ^a   Siegert, P ^a   Mesch, K ^a   Bruhn, H ^a   Grotzinger J ^b  

^a HEINRICH HEINE UNIVERSITY   (Germany)

^b UNIVERSITY HOSPITAL RWTH AACHEN   (Germany)

Author keywords

2 hydroxy ketone; Phenylacetylcarbinol; Pyruvate decarboxylase; Site directed mutagenesis; Thiamin diphosphate

Indexed keywords

COCARBOXYLASE; MAGNESIUM; METHANOL DERIVATIVE; PYRUVATE DECARBOXYLASE;

ARTICLE; BACTERIUM MUTANT; CARBOXYLATION; CATALYSIS; ENZYME ACTIVE SITE; ENZYME ACTIVITY; ENZYME DEGRADATION; ENZYME STABILITY; NONHUMAN; PRIORITY JOURNAL; SITE DIRECTED MUTAGENESIS; STEREOCHEMISTRY; ZYMOMONAS MOBILIS;

BACTERIA (MICROORGANISMS); ZYMOMONAS MOBILIS;

EID: 0032213506     PISSN: 00142956     EISSN: None     Source Type: Journal    
DOI: 10.1046/j.1432-1327.1998.2570538.x     Document Type: Article

References (31)

1. Harris, T. K. & Washabaugh, M. W. (1995) Distribution of the thiamin diphosphate C(2)-proton during catalysis of acetaldehyde formation by brewers' yeast pyruvate decarboxylase, Biochemistry 34, 13994-14000.
2. Sun, S. X., Duggleby, R. G. & Schowen, R. L. (1995) Linkage of catalysis and regulation in enzyme action - carbon isotope effects, solvent isotope effects, and proton inventories for the unregulated pyruvate decarboxylase of Zymomonas mobilis, J. Am. Chem. Soc. 117, 7317-7322.
3. Alvarez, F. J., Ermer, J., Hübner, G., Schellenberger, A. & Schowen, R. L. (1995) The linkage of catalysis and regulation in enzyme action. Solvent isotope effects as probes of protonic sites in the yeast pyruvate decarboxylase mechanism, J. Am. Chem. Soc. 117, 1678-1683.
4. Kern, D., Kern, G., Neef, H., Tittmann, K., Killenberg-Jabs, M., Wikner, C., Schneider, G. & Hübner, G. (1997) How thiamine diphosphate is activated in enzyme, Science 275, 67-70.
5. Dyda, F., Furey, W., Swaminathan, S., Sax, M., Farrenkopf, B. C. & Jordan, F. (1993) Catalytic center in the thiamin diphosphate dependent enzyme pyruvate decarboxylase at 2.4-Å resolution, Biochemistry 32, 6165-6170.
6. Arjunan, P., Umland, T., Dyda, F., Swaminathan, S., Furey, W., Sax, M., Farrenkopf, B., Gao, Y., Zhang, D. & Jordan, F. (1996) Crystal structure of the thiamin diphosphate-dependent enzyme pyruvate decarboxylase from the yeast Saccharomyces cerevisiae at 2.3 Å resolution, J. Mol. Biol. 256, 590-600.
7. Lu, G., Dobritzsch, D., König, S. & Schneider, G. (1997) Novel tetramer assembly by pyruvate decarboxylase from brewer's yeast observed in a new crystal form, FEBS Lett. 403, 249-253.
8. Jordan, F., Barburina, I., Gao, Y., Kahyaoglu, A., Nemeria, N., Volkov, A., Yi, J. Z., Zhang, D., Machado, R., Guest, J. R., Furey, W., Hohmann, S. (1996) New insights to the regulation of thiamin diphosphate decarboxylases by substrate and ThDP. Mg(II), in Biochemistry and physiology of thiamin diphosphate dependent enzymes (Bisswanger H. & Schellenberger, A., eds) p. 53, A. & C. Intemann, Wissenschaftlicher Verlag, Prien.
9. Schenk, G., Leeper, F. J., England, R., Nixon, P. F. & Duggleby, R. G. (1997) The role of His113 and His114 in pyruvate decarboxylase from Zymomonas mobili, Eur. J. Biochem. 248, 63-71.
10. Harris, T. K. & Washabaugh, M. W. (1995) Solvent-derived protons in catalysis by brewers' yeast pyruvate decarboxylase, Biochemistry 34, 14001-14011.
11. Lobell, M. & Crout, D. H. G. (1996) Pyruvate decarboxylase: a molecular modeling study of pyruvate decarboxylation and acyloin formation, J. Am. Chem. Soc. 118, 1867-1873.
12. Lobell, M. & Crout, D. H. G. (1996) New insight into the pyruvate decarboxylase-catalysed formation of lactaldehyde from H-D exchange experiments: a 'water proof' active site, J. Chem. Soc. Perkin Trans. 1, 1577-1581.
13. Bornemann, S., Crout, D. H. G., Dalton, H., Kren, V., Lobell, M., Dean, G., Thomson, N. & Turner, M. M. (1996) Stereospecific formation of R-aromatic acyloins by Zymomonas mobilis pyruvate decarboxylase, J. Chem. Soc. Perkin Trans. 1, 425-430.
14. Mesch, K. (1997) Rationales Protein-Design an der Pyruvatdecarboxylase aus Zymomonas mobilis, Doctoral thesis, University of Düsseldorf.
15. Reynen, M. & Sahm, H. (1988) Comparison of the structural genes of pyruvate decarboxylases in different Zymomonas mobilis strains, J. Bacteriol. 170, 3310-3313.
16. Bruhn, H., Pohl, M., Grötzinger, J. & Kula, M. R. (1995) The replacement of Trp392 by alanine influences the decarboxylase/carboligase activity and stability of pyruvate decarboxylase from Zymomonas mobilis, Eur. J. Biochem. 234, 650-655.
17. Ho, S. N., Hunt, H. D., Horton, R. M., Pullen, J. K. & Pease, L. R. (1989) Site-directed mutagenesis by overlap extension using the polymerase chain reaction, Gene (Amst.) 77, 51-59.
18. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989) Molecular cloning: a laboratory manual, 2nd edn, Cold Spring Harbor Laboratory, Cold Spring Harbor NY.
19. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriaphage T4, Nature 227, 680-685.
20. Kyse-Anderson, J. (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose, J. Biochem. Biophys. Methods 10, 203-209.
21. Bradford, M. M. (1976) A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of protein-binding dye, Anal. Biochem. 72, 248-254.
22. Pohl, M., Grötzinger, J., Wollmer, A. & Kula, M. R. (1994) Reversible dissociation and unfolding of pyruvate decarboxylase from Zymomonas mobilis, Eur. J. Biochem. 224, 651-661.
23. Pohl, M., Mesch, K., Rodenbrock, A. & Kula, M. R. (1995) Stability investigations on the pyruvate decarboxylase from Zymomonas mobilis, Biotechnol. Appl. Biochem. 22, 95-105.
24. Gröger, G. & Erge, D. (1964) Zur Analytik des Phenylacetylcarbinols, eines Zwischenprodukts bei der Ephedrinsynthese, Pharmazie 20, 92-95.
25. Crout, D. H. G. & Morrey, S. M. (1983) Synthesis of (R)-and (S)-acetoin (3-hydroxybutan-2-one), J. Chem. Soc. Perkin Trans. 1, 2435-2440.
26. Koga, J., Adachi, T. & Hidaka, H. (1991) Molecular cloning of the gene for indolpyruvate decarboxylase from Enterobacter cloacae, Mol. Gen. Genet. 226, 1-6.
27. Tsou, A. Y., Ransom, S. C., Gerlt, J. A., Buechter, D. D., Babbitt, P. C. & Kenyon, G. L. (1990) Mandelate pathway of Pseudomonas putida: sequence relationship involving mandelate racemase, (S)-mandelate dehydrogenase, and benzoylformate decarboxylase and expression of benzoylformate decarboxylase in Escherichia coli, Biochemistry 29, 9856-9862.
28. Hasson, M. S., Muscate, A., Henehan, G. T. M., Guidinger, P. F., Petsko, G. A., Ringe, D. & Kenyon, G. L. (1995) Purification and crystallization of benzoylformate decarboxylase, Protein Sci. 4, 955-959.
29. Iding, H., Siegert, P., Mesch, K. & Pohl., M. (1998) Application of α-keto acid decarboxylases in biotransformations, Biochim. Biophys, Acta 1385, 307-322.
30. Hawkins, C. F., Borges, A. & Perham, R. N. (1989) A common structural motif in thiamin pyrophosphate-hinding enzymes, FEBS Lett. 255, 77-82.
31. Pohl, M. (1997) Protein design on pyruvate decarboxylase (PDC) by site-directed mutagenesis, Adv. Biochem. Eng. Biotechnol. 58, 16-43.