ZgGlc16A（endo-β-1,3-Glucanase）
ZgGlc16A
Ex-Glu0312
(EC.3.2.1.39) endo-β-1,3-Glucanase
CAZy Family: GH16
PROPERTIES
1. ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~34kDa)
Figure 1. Electrophoresis analysis of ZgGlc16A. 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, ZgGlc16A purified from Ni sepharose fastflow column.
2. SPECIFIC ACTIVITY
42.3 U/mg protein (on Curdlan) at pH 7.0 and 40°C
One Unit of β-glucan activity is defined as the amount of enzyme required to release 1 μmol of glucose from Curdlan (10 mg/mL) in phosphate buffer(200 mM) pH7.0.
3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of ZgGlc16A on different substrates.
Substrate |
Linkage |
ZgGlc16A |
|
Specific activit（U/mg） |
Relative activity (%) |
||
curdlan |
β-1,3 |
42.3 |
100 |
ganoderan |
β-1,3 |
27.5 |
65 |
barley polysaccharide |
β-1,3/1,4 |
14.2 |
34 |
Cellulose polysaccharide |
β-1,4 |
0 |
0 |
Shier polysaccharide |
β-1,6 |
0 |
0 |
starch polysaccharide |
α-1,4 |
0 |
0 |
Reactions were performed with 10 mg/ml (polysaccharides) substrate, pH 7.0, at 40°C for 5min.
Fig.2. HPAEC detection results of ZgGlc16A action site study. The experiment used gel polysaccharide as substrate. a, monosaccharide. b, disaccharide. c, trisaccharide.
Fig.3. HPAEC diagram of barley polysaccharide hydrolyzed by ZgGlc16A.G1, monosaccharide.G2, Linear glucobiose.G3, Linear glucotriose.G4, Linear glucotetraose.
Fig.4. HPAEC diagram of ZgGlc16A after hydrolysis of Ganoderma lucidum polysaccharides. G1, monosaccharide.G2, Linear glucobiose.G3, Linear glucotriose.G4, Linear glucotetraose.
4. PHYSICOCHEMICAL PROPERTIES
pH Optima: 6.0
pH Stability: 6.0-8.0
Temperature Optima: 40°C
Temperature Stability: <30°C
5. STORAGE CONDITIONS
The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in phosphate buffer (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.
6. REFERENCES
[1]邓艳楠. 食半乳聚糖卓贝尔氏黄杆菌中两种新的β-1,3-葡聚糖酶的重组表达及功能研究[D]. 东北师范大学, 2023.
