BtGlu30A

Ex-Glu0011

(EC.3.2.1.75)endo-β-1,6-Glucanase

CAZy Family: GH30

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~55kDa)

Figure 1. Electrophoresis analysis of BtGlu30A. 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, BtGlu30A purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

235 U/mg protein (on yeast β-glucan) at pH 7.5 and 37°C

One Unit of β-glucan activity is defined as the amount of enzyme required to release 1 μg of glucose from yeast β-glucan (10mg/mL) in 60 min.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Kinetic parameters for BtGlu30A on different substrates.

^aThe kinetic parameter is mg ml^-1min^-1for pustulan and M^-1min^-1for glucooligosaccharides.

Figure 2. HPAEC-PAD analysis of BtGlu30A.BtGlu30A(50 nM) was incubated with 2 mg ml^-1 1, 6-β-glucan at 37 °C. Samples were taken at the indicated time points and analyzed by HPAEC-PAD.

4.PHYSICOCHEMICAL PROPERTIES

Recommend pH:7.0

Recommend Temperature:37°C

5.STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in phosphate buffer (10 mM) pH7.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1]Max J. Temple, Fiona Cuskin, Arnaud Baslé, et al. A Bacteroidetes locus dedicated to fungal 1,6-β-glucan degradation: Unique substrate conformation drives specificity of the key endo-1,6-β-glucanase[J].J.Biol.Chem. 2017,292(25):10639 –10650.