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Biochem J. Apr 1, 2001; 355(Pt 1): 155–165.
PMCID: PMC1221723

Pectate lyase 10A from Pseudomonas cellulosa is a modular enzyme containing a family 2a carbohydrate-binding module.


Pectate lyase 10A (Pel10A) enzyme from Pseudomonas cellulosa is composed of 649 residues and has a molecular mass of 68.5 kDa. Sequence analysis revealed that Pel10A contained a signal peptide and two serine-rich linker sequences that separate three modules. Sequence similarity was seen between the 9.2 kDa N-terminal module of Pel10A and family 2a carbohydrate-binding modules (CBMs). This N-terminal module of Pel10A was shown to encode an independently functional module with affinity to crystalline cellulose. A high sequence identity of 66% was seen between the 14.2 kDa central module of Pel10A and the functionally uncharacterized central modules of the xylan-degrading enzymes endoxylanase 10B, arabinofuranosidase 62C and esterase 1D, also from P. cellulosa. The 35.8 kDa C-terminal module of Pel10A was shown to have 30 and 36% identities with the family 10 pectate lyases from Azospirillum irakense and an alkaliphilic strain of Bacillus sp. strain KSM-P15, respectively. This His-tagged C-terminal module of the Pel10A was shown to encode an independent catalytic module (Pel10Acm). Pel10Acm was shown to cleave pectate and pectin in an endo-fashion and to have optimal activity at pH 10 and in the presence of 2 mM Ca2+. Highest enzyme activity was detected at 62 degrees C. Pel10Acm was shown to be most active against pectate (i.e. polygalacturonic acid) with progressively less activity against 31, 67 and 89% esterified citrus pectins. These data suggest that Pel10A has a preference for sequences of non-esterified galacturonic acid residues. Significantly, Pel10A and the P. cellulosa rhamnogalacturonan lyase 11A, in the accompanying article [McKie, Vincken, Voragen, van den Broek, Stimson and Gilbert (2001) Biochem. J. 355, 167-177], are the first CBM-containing pectinases described to date.

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Selected References

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  • Black GW, Rixon JE, Clarke JH, Hazlewood GP, Theodorou MK, Morris P, Gilbert HJ. Evidence that linker sequences and cellulose-binding domains enhance the activity of hemicellulases against complex substrates. Biochem J. 1996 Oct 15;319(Pt 2):515–520. [PMC free article] [PubMed]
  • Fuchs A. The trans-eliminative breakdown of Na-polygalacturonate by Pseudomonas fluorescens. Antonie Van Leeuwenhoek. 1965;31(3):323–340. [PubMed]
  • McCarter JD, Withers SG. Mechanisms of enzymatic glycoside hydrolysis. Curr Opin Struct Biol. 1994 Dec;4(6):885–892. [PubMed]
  • Hazlewood GP, Gilbert HJ. Structure and function analysis of Pseudomonas plant cell wall hydrolases. Biochem Soc Trans. 1998 May;26(2):185–190. [PubMed]
  • McKie VA, Vincken JP, Voragen AG, van den Broek LA, Stimson E, Gilbert HJ. A new family of rhamnogalacturonan lyases contains an enzyme that binds to cellulose. Biochem J. 2001 Apr 1;355(Pt 1):167–177. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Moran F, Nasuno S, Starr MP. Extracellular and intracellular polygllacturonic acid trans-eliminases of Erwinia carotovora. Arch Biochem Biophys. 1968 Feb;123(2):298–306. [PubMed]
  • Fraaije BA, Bosveld M, Van den Bulk RW, Rombouts FM. Analysis of conductance responses during depolymerization of pectate by soft rot Erwinia spp. and other pectolytic bacteria isolated from potato tubers. J Appl Microbiol. 1997 Jul;83(1):17–24. [PubMed]
  • Bekri MA, Desair J, Keijers V, Proost P, Searle-van Leeuwen M, Vanderleyden J, Vande Broek A. Azospirillum irakense produces a novel type of pectate lyase. J Bacteriol. 1999 Apr;181(8):2440–2447. [PMC free article] [PubMed]
  • Ferreira LM, Hazlewood GP, Barker PJ, Gilbert HJ. The cellodextrinase from Pseudomonas fluorescens subsp. cellulosa consists of multiple functional domains. Biochem J. 1991 Nov 1;279(Pt 3):793–799. [PMC free article] [PubMed]
  • Hall J, Gilbert HJ. The nucleotide sequence of a carboxymethylcellulase gene from Pseudomonas fluorescens subsp. cellulosa. Mol Gen Genet. 1988 Jul;213(1):112–117. [PubMed]
  • Kellett LE, Poole DM, Ferreira LM, Durrant AJ, Hazlewood GP, Gilbert HJ. Xylanase B and an arabinofuranosidase from Pseudomonas fluorescens subsp. cellulosa contain identical cellulose-binding domains and are encoded by adjacent genes. Biochem J. 1990 Dec 1;272(2):369–376. [PMC free article] [PubMed]
  • Ferreira LM, Wood TM, Williamson G, Faulds C, Hazlewood GP, Black GW, Gilbert HJ. A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain. Biochem J. 1993 Sep 1;294(Pt 2):349–355. [PMC free article] [PubMed]
  • Sawada K, Ogawa A, Ozawa T, Sumitomo N, Hatada Y, Kobayashi T, Ito S. Nucleotide and amino-acid sequences of a new-type pectate lyase from an alkaliphilic strain of Bacillus. Eur J Biochem. 2000 Mar;267(5):1510–1515. [PubMed]
  • Daas PJ, Arisz PW, Schols HA, De Ruiter GA, Voragen AG. Analysis of partially methyl-esterified galacturonic acid oligomers by high-performance anion-exchange chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Anal Biochem. 1998 Mar 15;257(2):195–202. [PubMed]
  • Hall J, Hazlewood GP, Huskisson NS, Durrant AJ, Gilbert HJ. Conserved serine-rich sequences in xylanase and cellulase from Pseudomonas fluorescens subspecies cellulosa: internal signal sequence and unusual protein processing. Mol Microbiol. 1989 Sep;3(9):1211–1219. [PubMed]
  • Gilbert HJ, Hall J, Hazlewood GP, Ferreira LM. The N-terminal region of an endoglucanase from Pseudomonas fluorescens subspecies cellulosa constitutes a cellulose-binding domain that is distinct from the catalytic centre. Mol Microbiol. 1990 May;4(5):759–767. [PubMed]
  • Hall J, Black GW, Ferreira LM, Millward-Sadler SJ, Ali BR, Hazlewood GP, Gilbert HJ. The non-catalytic cellulose-binding domain of a novel cellulase from Pseudomonas fluorescens subsp. cellulosa is important for the efficient hydrolysis of Avicel. Biochem J. 1995 Aug 1;309(Pt 3):749–756. [PMC free article] [PubMed]
  • Poole DM, Hazlewood GP, Huskisson NS, Virden R, Gilbert HJ. The role of conserved tryptophan residues in the interaction of a bacterial cellulose binding domain with its ligand. FEMS Microbiol Lett. 1993 Jan 1;106(1):77–83. [PubMed]
  • Fanutti C, Ponyi T, Black GW, Hazlewood GP, Gilbert HJ. The conserved noncatalytic 40-residue sequence in cellulases and hemicellulases from anaerobic fungi functions as a protein docking domain. J Biol Chem. 1995 Dec 8;270(49):29314–29322. [PubMed]
  • Millward-Sadler SJ, Hall J, Black GW, Hazlewood GP, Gilbert HJ. Evidence that the Piromyces gene family encoding endo-1,4-mannanases arose through gene duplication. FEMS Microbiol Lett. 1996 Aug 1;141(2-3):183–188. [PubMed]
  • Halstead JR, Vercoe PE, Gilbert HJ, Davidson K, Hazlewood GP. A family 26 mannanase produced by Clostridium thermocellum as a component of the cellulosome contains a domain which is conserved in mannanases from anaerobic fungi. Microbiology. 1999 Nov;145(Pt 11):3101–3108. [PubMed]
  • Scavetta RD, Herron SR, Hotchkiss AT, Kita N, Keen NT, Benen JA, Kester HC, Visser J, Jurnak F. Structure of a plant cell wall fragment complexed to pectate lyase C. Plant Cell. 1999 Jun;11(6):1081–1092. [PMC free article] [PubMed]
  • McKie VA, Black GW, Millward-Sadler SJ, Hazlewood GP, Laurie JI, Gilbert HJ. Arabinanase A from Pseudomonas fluorescens subsp. cellulosa exhibits both an endo- and an exo- mode of action. Biochem J. 1997 Apr 15;323(Pt 2):547–555. [PMC free article] [PubMed]
  • Braithwaite KL, Barna T, Spurway TD, Charnock SJ, Black GW, Hughes N, Lakey JH, Virden R, Hazlewood GP, Henrissat B, et al. Evidence that galactanase A from Pseudomonas fluorescens subspecies cellulosa is a retaining family 53 glycosyl hydrolase in which E161 and E270 are the catalytic residues. Biochemistry. 1997 Dec 9;36(49):15489–15500. [PubMed]
  • Tardy F, Nasser W, Robert-Baudouy J, Hugouvieux-Cotte-Pattat N. Comparative analysis of the five major Erwinia chrysanthemi pectate lyases: enzyme characteristics and potential inhibitors. J Bacteriol. 1997 Apr;179(8):2503–2511. [PMC free article] [PubMed]
  • Liu Y, Chatterjee A, Chatterjee AK. Nucleotide sequence and expression of a novel pectate lyase gene (pel-3) and a closely linked endopolygalacturonase gene (peh-1) of Erwinia carotovora subsp. carotovora 71. Appl Environ Microbiol. 1994 Jul;60(7):2545–2552. [PMC free article] [PubMed]

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