BtMan92K

Ex-Man0276

(EC3.2.1)exo-α-1,3-mannosidase

CAZy Family: GH92

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~83 kDa)

Figure 1. Electrophoresis analysis of BtMan92K. 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, BtMan92K purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

6.46 U/mg protein (on pNP-α-man) at pH 7 and 37°C.

One Unit of pNP-α-manactivity is defined as the amount of enzyme required to release 1 μmol of pNP per minute from pNP-α-man (5 mM) in PBS buffer (10 mM) pH 7.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtMan92K on different substrates^a.

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

^bAbsorption caused by released p-nitrophenol or reducing sugars was measured at 405 nm. The relative activity on pNP-α-Man was taken as 100%.

4.STORAGE CONDITIONS

The enzyme is supplied in PBS buffer and should be stored at -20°C. For assay, this enzyme should be diluted in PBS buffer pH 7.0. Swirl to mix the enzyme immediately prior to use.