BoRgl11A （Rhamnogalacturonan lyase）
BoRgl11A
En-Rgl0182
(EC. 4.2.2.23) Rhamnogalacturonan lyases
CAZy Family:PL11
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~68 kDa)
Fig 1. Electrophoresis analysis of BoRgl11A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of E. coliBL21-pET28a-borgl11a before IPTG induction; lane 2, culture lysate of E. coliBL21-pET28a-borgl11a after IPTG induction; lane 3, BoRgl11A purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
3.9 U/mg protein (on RG-I-AT from Adenophora tetraphylla(Thunb.)Fisch.) at pH7.0 and 30°C
One unit of RGL activity was defined as the amount of enzyme required to generate 1 mmol of 4,5-unsaturated galacturonic acid in 1 min.
3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1 Substrate specificity ofBoRgl11A
Substrate |
Structural features |
Relative activity (%) |
De-HG |
HG |
16.2±2.2 |
HG (48% Methyl-esterified) |
HG |
-- |
RG-II |
RG-II |
-- |
RG-I-AT |
RG-I |
100.0±2.0 |
RG-I-AT-PP |
RG-I backbone |
135.6±3.5 |
RG-I-P |
RG-I+HG |
35.7±2.8 |
Galactan |
Galactan+RG-I backbone |
30.8±1.6 |
Arabinan |
Arabinan+RG-I backbone |
14.5±0.7 |
4.PHYSICOCHEMICAL PROPERTIES
pH Optima:9.0
pH Stability:7.0-11.0
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.
