PpRgl11B （Rhamnogalacturonan lyase）
PpRgl11B
En-Rgl0188
(EC.4.2.2.23) Rhamnogalacturonan lyase
CAZy Family:PL11
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~88kDa)
Fig1.Electrophoresis analysis of PpRgl11B. M, molecular weight marker(PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of E. coliBL21-pET28a-pprgl11b before IPTG induction; lane 2, culture lysate of E. coliBL21-pET28a-pprgl11b after IPTG induction; lane 3, PpRgl11B purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
6.0 U/mg protein (on RG-I-AT from Adenophora tetraphylla(Thunb.)Fisch.) at pH7.0 and 30oC
One unit of RGL activity was defined as the amountof enzyme required to generate 1 μmol of 4,5-unsaturatedgalacturonic acid in 1 min.
3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1 Substrate specificity of BoRgl11B
Substrate |
Structural features |
Relative activity (%) |
De-HG |
HG |
5.9±0.7 |
HG (48% Methyl-esterified) |
HG |
7.5±0.5 |
RG-II |
RG-II |
-- |
RG-I-AT |
RG-I |
100.0±2.8 |
RG-I-AT-PP |
RG-I backbone |
110.4±2.6 |
RG-I-P |
RG-I+HG |
47.2±1.6 |
Galactan |
Galactan+RG-I backbone |
17.2±0.9 |
Arabinan |
Arabinan+RG-I backbone |
3.6±0.4 |
4.PHYSICOCHEMICAL PROPERTIES
pH Optima:8.0
pH Stability:6.0-9.0
Temperature Optima:30°C
Temperature Stability: <25°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.
