CtRgl11A（Rhamnogalacturonan lyase）
CtRgl11A
En-Rgl0192
(EC. 4.2.2.23) Rhamnogalacturonan lyase
CAZy Family: PL11
PROPERTIES
1. ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~80 kDa)
Fig 1. Electrophoresis analysis of CtRgl11A. 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, CtRgl11A purified from Ni sepharose fast flow column.
2. SPECIFIC ACTIVITY
5.4 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 mmol of 4,5-unsaturated galacturonic acid in 1 min.
3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1 Activity of BtRgl11A
Substrate |
Specific activity (U/mg) |
Km (mg) |
kcat (s-1) |
kcat/Km(mg-1s-1) |
Rhamnogalacturonan (Soybean) |
9.8 |
4.8 |
21.2 |
4.41 |
Rhamnogalacturonan (Potato) |
9.1 |
4.9 |
21.0 |
4.28 |
Pectic galactan (Lupin) |
4.7 |
10 |
7.7 |
0.77 |
Galactan (Potato) |
3.8 |
4.7 |
6.4 |
1.36 |
Pectin (25 % methyl-esterified) |
4.3 |
8.2 |
9.8 |
1.19 |
Polygalacturonic acid |
1.2 |
ND |
ND |
ND |
4. PHYSICOCHEMICAL PROPERTIES
pH Optima: 8.5
Temperature Optima: 70°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 8.0. Swirl to mix the enzyme immediately prior to use.
6. REFERENCE
[1] Dhillon A, Fernandes V O, Dias F, et al. A new member of family 11 polysaccharide lyase, rhamnogalacturonan lyase (CtRGLf) from Clostridium thermocellum[J]. Molecular biotechnology, 2016, 58(4): 232-240.
