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)

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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^a.

Substrate	Linkage	ZgGlc16A
Substrate	Linkage	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

^a Reactions were performed with 10 mg/ml (polysaccharides) substrate, pH 7.0, at 40°C for 5min.

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Fig.2. HPAEC detection results of ZgGlc16A action site study. The experiment used gel polysaccharide as substrate. a, monosaccharide. b, disaccharide. c, trisaccharide.

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Fig.3. HPAEC diagram of barley polysaccharide hydrolyzed by ZgGlc16A.G1, monosaccharide.G2, Linear glucobiose.G3, Linear glucotriose.G4, Linear glucotetraose.

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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.

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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)

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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^a.

Substrate	Linkage	ZgGlc16A
Substrate	Linkage	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

^a Reactions were performed with 10 mg/ml (polysaccharides) substrate, pH 7.0, at 40°C for 5min.

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Fig.2. HPAEC detection results of ZgGlc16A action site study. The experiment used gel polysaccharide as substrate. a, monosaccharide. b, disaccharide. c, trisaccharide.

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Fig.3. HPAEC diagram of barley polysaccharide hydrolyzed by ZgGlc16A.G1, monosaccharide.G2, Linear glucobiose.G3, Linear glucotriose.G4, Linear glucotetraose.

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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.