PcRgl4A

En-Rgl0175 

(EC. 4.2.2.23)  Rhamnogalacturonan lyase

CAZy Family: PL4

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~76 kDa)

Figure 1. Electrophoresis analysis of PcRgl4A. 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, PcRgl4A purified from Ni sepharose fast flow column.

2. SPECIFIC ACTIVITY

1.8 U/mg protein (on RG-I-AT from Adenophora tetraphylla(Thunb.)Fisch.) at pH8.0 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

Figure 2. PcRGL4A degrading activity against soybean RG (closed circles) and sugar beet HG (open circles) .

Figure 3. HPAEC (a) and MALDI-TOF MS (b) analyses of the enzymatic products of RG-AG incubated with PcRgl4A.

Figure 4. HILIC-UV (UV235 nm) chromatograms of PcRgl4A oligosaccharides.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 40°C

5. STORAGE CONDITIONS

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

6. REFERENCE

[1] Iwai M, Yamada H, Ikemoto T, et al. Biochemical characterization and overexpression of an endo-rhamnogalacturonan lyase from Penicillium chrysogenum. Molecular Biotechnology, 2015, 57(6): 539-548.