BtGlu3A

Ex-Glu0012

(EC.3.2.1.21) exo-β-1,6-Glucosidase

CAZy Family: GH3

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~85 kDa)

Figure 1. Electrophoresis analysis of BtGlu3A. 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, BtGlu3A purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

24 U/mg protein (on pNP-β-glu) at pH 7.0 and 37°C

One Unit of pNP-β-glu activity is defined as the amount of enzyme required to release one μmole of glucose per minute from pNP-β-glu (5 mM) in phosphate buffer (10 mM) pH 7.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1.Kinetic parameters for BtGlu3A on different substrates.

^apNP-β-D-Glc, 4-nitrophenyl β-D-glucopyranoside.

^bND, not determined.

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) pH 7.0. 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. Biol. Chem. 2017, 292(25): 10639 –10650.