EcRgl4A

En-Rgl0179

(EC. 4.2.2.23) Rhamnogalacturonan lyase

CAZy Family: PL4

PROPERTIES

1.  ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~61 kDa)

Figure 1. Electrophoresis analysis of EcRgl4A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate before IPTG induction; lane 2, culture lysate after IPTG induction; lane 3, EcRgl4A purified from Ni sepharose fast flow column.

2. SPECIFIC ACTIVITY

4.3 U/mg protein (on RG-I-AT from Adenophora tetraphylla(Thunb.)Fisch.) at pH6.0 and 37°C.

One unit of RGL activity was defined as the amount of enzyme required to generate 1 μmol of 4,5-unsaturated galacturonic acid in 1 min.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Several substrates (polygalacturonate, pectin, RG-I, and arabinogalactan) in the presence of EcRgl4A. We could detect activity only with RG-I, confirming that EcRgl4A is a rhamnogalacturonate lyase which cleaves the RG-I backbone between a rhamnose and a galacturonate and creates an unsaturated bond between carbons 4 and 5 of the galacturonate residue.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 6.0

Temperature Optima: 37°C

5. STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in phosphate buffer (50 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.

6. References

[1] Laatu M, Condemine G. Rhamnogalacturonate lyase RhiE is secreted by the out system in Erwinia chrysanthemi. Journal of bacteriology, 2003, 185(5): 1642-1649.