CcGlu1B

Ex-Glu0009

(EC.3.2.1.195) exo-β-Glucosidase

CAZy Family: GH1

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~57 kDa)

Figure 1. Electrophoresis analysis of CcGlu1B. 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, CcGlu1B purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

1.1 U/mg protein (on pNP-β-glu) at pH 6.0 and 37°C

One Unit of pNP-β-glu activity is defined as the amount of enzyme required to release 1 μmol of glucose per minute from pNP-β-glu (5 mM) in phosphate buffer (50 mM) pH 6.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of CcGlu1B on different substrates^a.

^aReactions were performed with 1 mM (p-nitrophenyl glycosides and disaccharides) or 5 mg/ml (polysaccharides) substrate, pH 6.0, at 37°C for 10 min.

^bThe relative activity on pNPbGlc and cellobiose were defined as 100%.

^cThe data are reported as means ± SD from the mean for three independent experiments.

^dNot detected.

Figure 2. IC analysis of hydrolytic products released from G2(a), G3(b), G4(c) and G5(d) by purifiedCcGlu1B. Reactions (500 μl) containing 1 mg/ml substrate were incubated at 30^oC in 50 mM phosphate buffer (pH 6.0) with 0.02 mg/ml CcBgl1B for the indicated times.

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 6.0

pH Stability: 5.5-10.0

Temperature Optima: 5°C

Temperature Stability:<40°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 7.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Yuan Y, Xu F, Yao J, et al. Cloning, expression and biochemical characterization of a GH1 β-glucosidase from Cellulosimicrobium cellulans. Biocatalysis and Biotransformation, 2018, 36(5): 1-10.