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Polysaccharide Lyase Family 20

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Revision as of 04:09, 7 August 2014 by Naotake Konno (talk | contribs)
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Polysaccharide Lyase Family PL20
Mechanism β-elimination
Metal Cofactor calcium
Active site residues unknown
CAZy DB link
http://www.cazy.org/PL20.html

Substrate specificities

Polysaccharide lyases of family 20 cleave β-1,4 linkages in polyglucuronate (β-1,4-glucuronan lyase; EC 4.2.2.14). The first PL20 enzyme was cloned from filamentous fungus Trichoderma reesei (TrGL) [1]. TrGL was highly specific for β-1,4-glucuronan prepared from regenerated cellulose by 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO) mediated oxidation (cellouronate).

Kinetics and Mechanism

A PL20 enzyme, TrGL, has been characterized as a β-1,4-glucuronan lyase. TrGL catalyzed depolymerization of β-1,4-glucuronan endolytically by β-elimination [1] The enzyme was most active at pH 6.5 and 50 °C, and its activity and thermostability increased in the presence of calcium ions.

Catalytic Residues

There are approximately 40 completely conserved amino-acid residues in the PL20 members [2] Possible catalytic residues has been predicted based on structural comparison between TrGL and PL7 alginate lyase A1–II’ [2] [3]. It seemed to be that the charge neutralizer, the catalytic base and the catalytic acid in the TrGL are Gln91, His53 and Tyr200, respectively. However, in order to clarify the substrate recognition mechanism and the identity of the catalytic residues of PL20, further studies will be required.

Three-dimensional structures

The structure of the TrGL was the first PL20 structure to be reported [2]. TrGL has a typical β-jelly roll fold (2ZZJ, 1.8 Å).

Family Firsts

First First catalytic activity
TrGl from Trichoderma reesei [1]
First 3-D structure
TrGl from Trichoderma reesei [2]

References

  1. Konno N, Igarashi K, Habu N, Samejima M, and Isogai A. (2009). Cloning of the Trichoderma reesei cDNA encoding a glucuronan lyase belonging to a novel polysaccharide lyase family. Appl Environ Microbiol. 2009;75(1):101-7. DOI:10.1128/AEM.01749-08 | PubMed ID:18978091 [Konno2009a]
  2. Konno N, Ishida T, Igarashi K, Fushinobu S, Habu N, Samejima M, and Isogai A. (2009). Crystal structure of polysaccharide lyase family 20 endo-beta-1,4-glucuronan lyase from the filamentous fungus Trichoderma reesei. FEBS Lett. 2009;583(8):1323-6. DOI:10.1016/j.febslet.2009.03.034 | PubMed ID:19306878 [Konno2009b]
  3. Ogura K, Yamasaki M, Mikami B, Hashimoto W, and Murata K. (2008). Substrate recognition by family 7 alginate lyase from Sphingomonas sp. A1. J Mol Biol. 2008;380(2):373-85. DOI:10.1016/j.jmb.2008.05.008 | PubMed ID:18514736 [Ogura2008]

All Medline abstracts: PubMed