BtMan125B

Ex-Man0097

(EC.3.2.1.163)exo-α-1,6-Mannosidase

CAZy Family: GH125

PROPERTIES

1.ELECTROPHORETIC PURITY

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

Figure 1. Electrophoresis analysis of BtMan125B. 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

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtMan125B 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 was measured at 405 nm.

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

Figure.2 BT2632 and BtMan125B at 100 nM wereincubated with 1 mg/ml of the debranched mannan for 1 h in the buffer. TLC analysis (a) and HPAEC traces (b) of thereactions areshown.

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.

6. REFERENCES

[1]Cuskin F, Lowe E C, Temple M J, et al. Human gut Bacteroidetes can utilize yeast mannan through a selfish mechanism[J]. Nature, 2015, 517(7533): 165-169.