BoRgl11B （Rhamnogalacturonan lyase）
BoRgl11B
En-Rgl0183
(EC.4.2.2.23) Rhamnogalacturonan lyases
CAZy Family:PL11
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~75kDa)
Fig1.Electrophoresis analysis of BoRgl11B. M, molecular weight marker(PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of E. coli BL21-pET28a-borgl11b before IPTG induction; lane 2, culture lysate of E. coliBL21-pET28a-borgl11b after IPTG induction; lane 3,BoRgl11B purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
1.1 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 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 ofBoRgl11B
Substrate |
Structural features |
Relative activity (%) |
De-HG |
HG |
15.6±2.1 |
HG (48% Methyl-esterified) |
HG |
7.4±1.9 |
RG-II |
RG-II |
-- |
RG-I-AT |
RG-I |
100.0±1.4 |
RG-I-AT-PP |
RG-I backbone |
150.6±6.8 |
RG-I-P |
RG-I+HG |
50.3±1.3 |
Galactan |
Galactan+RG-I backbone |
1.3±0.2 |
Arabinan |
Arabinan+RG-I backbone |
1.9±0.3 |
4.PHYSICOCHEMICAL PROPERTIES
pH Optima:6.0
pH Stability:5.5-7.5
Temperature Optima:45°C
Temperature Stability: <25°C
5.STORAGE CONDITIONS
The enzymeshould 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.
