BtMan76A

Ex-Man0099

(EC.3.2.1.163)exo-α-1,6-Mannosidase

CAZy Family: GH76

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~48 kDa)

Figure 1. Electrophoresis analysis ofBtMan76A. 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, BtMan125B purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

30 U/mg protein (on pNP-α-Man) at pH 7.0 and 37°C.

One Unit of pNP-α-Man activity is defined as the amount of enzyme required to release 1 nmol of p-nitrophenyl per minute from  pNP-α-Man (5 mM) in PBS buffer (20 mM) pH 7.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtMan76A on different substrates^a.

^aReactions were performed with 5mM substrate, pH 7.0, at 37°C for 10 min.

^bAbsorption caused by releasedp-nitrophenol was measured at 405 nm.

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

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 37°C

5.STORAGE CONDITIONS

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