BvRgl11A （Rhamnogalacturonan lyase）
BvRgl11A
En-Rgl0184
(EC.4.2.2.23) Rhamnogalacturonan lyases
CAZy Family:PL11
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~68kDa)
Fig1.Electrophoresis analysis of BvRgl11A. M, molecular weight marker(PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of E. coliBL21-pET28a-bvrgl11a before IPTG induction; lane 2, culture lysate of E. coliBL21-pET28a-bvrgl11a after IPTG induction; lane 3,BvRgl11A purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
4.2U/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 amount of enzyme required to generate 1 mmol 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 |
7.2±0.7 |
HG (48% Methyl-esterified) |
HG |
9.0±0.1 |
RG-II |
RG-II |
1.3±0.2 |
RG-I-AT |
RG-I |
100.0±3.2 |
RG-I-AT-PP |
RG-I backbone |
125.6±5.3 |
RG-I-P |
RG-I+HG |
61.6±5.5 |
Galactan |
Galactan+RG-I backbone |
20.5±2.3 |
Arabinan |
Arabinan+RG-I backbone |
2.9±1.5 |
4.PHYSICOCHEMICAL PROPERTIES
pH Optima:9.0
pH Stability:7.0-10.0
Temperature Optima:35°C
Temperature Stability: <30°C
5.STORAGE CONDITIONS
The enzyme should be stored at-20°C. For assay, this enzyme should be diluted inphosphate buffer (50 mM) pH7.0. Swirl to mix the enzyme immediately prior to use.
