Erwinia chrysanthemi pectate lyase from

Figure 6. HPLC kinetics of polygalacturonic acid depolymerization by extracellular pectate lyases from crude supernatants of Erwinia chrysanthemi and Lachnospira multiparus cultures. A panels are full scale representations of products found over the reaction time sequence. B panels have expanded ordinates to better demonstrate the kinetics of minor products. Area unit refers to integration from HPLC tracings of product absorbance at 235 nm. Numbers in the panels refer to the degree of polymerization for individual products. Conditions for enzyme assay and product detection were the same as described for Figure 5.

The Family 9 pectate lyase from the plant pathogen Erwinia chrysanthemi has again a structure composed largely of (3-strands, being a 10-turn helix of parallel (3-strands. Analogy with the closely similar PL 1 enzymes suggests that the enzyme binds a catalytic Ca " between enzyme and substrate, in addition to... 

Lietzke, S.E., Yoder, M.D., Keen, N.T. and Jurnak, F. (1994) The three-dimensional structure of pectate lyase E, a plant virulence factor from Erwinia chrysanthemi. Plant Physiol Q6, 849-862. 

The first structures of this kind were reported in 1993 pectate lyase G from Erwinia chrysanthemi (Yoder et al, 1993) and alkaline protease from Pseudomonas aeruginosa (Baumann et al, 1993). Based on consideration of these crystal structures, the term parallel //-helix was introduced for a fold containing three //-strands per coil, and parallel //-roll for a fold with two //-strands per coil (Baumann etal, 1993 Yoder andjurnak, 1995 Yoder et al., 1993). The epithet parallel was intended to emphasize the distinction between these folds and the previously observed helical structure of the antibiotic gramicidin which contains both l- and D-amino acids and... 

Dehdashti, S.J., Doan, C. N., Chao, K. L., and Yoder, M. D. (2003). Effect of mutations in the T1.5 loop of pectate lyase A from Erwinia chrysanthemi EC16. Acta Crystallogr. D Biol. Crystallogr. 59, 1339—1342. 

Jenkins, J., Shevchik, V. E., Hugouvieux-Cotte-Pattat, N., and Pickersgill, R. W. (2004). The crystal structure of pectate lyase Pel9A from Erwinia chrysanthemi. J. Biol. Chem. 279, 9139-9145. 

Figure 5. HPLC profiles of products from reactions catalyzed by secreted pectate lyase activities from Erwinia chrysanthemi after 66 min and 192 min. Reaction mixtures contained 4 units of enzyme activity per ml and 0.1% PGA in 0.05 M Tris-HCl, pH 8.5, 0.2 mM CaClg. Injections of 0.05 ml were made from reaction mixtures with a WISP automatic sample injector and eluted at 2.0 ml/min with a run time of 60 min. Compounds eluting with retention times of 5.36, 6.40, 7.76, and 9.56 min corresponded to unsaturated oligogalacturonate reference standards with DP values of 2, 3, 4, and 5, respectively.

Figure 7. HPLC kinetic profiles of reaction products of pectate lyases obtained from chromatofocused fractions of Erwinia chrysanthemi culture supernatants. Chromatofocused enzymes, eluted at pH 8.6 (A), pH 8.3 (B), pH 6.0 (C), and < pH 6.0 (D), were assayed under conditions similar to those described in Figure 5.

Castang S, Shevchik VE, Hugouvieux-Cotte-Pattat N et al. (2(X)4) Ctystallization of the pectate lyase Pell from Erwinia chrysanthemi and SAD phasing of a gold derivative. Acta Cryst 60 190-... 

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