BtMan92G

Ex-Man0095

(EC.3.2.1.77)exo-α-1,2/3-Mannosidases

CAZy Family: GH92

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~87kDa)

Figure 1. Electrophoresis analysis of BtMan92G. 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, BtMan92Gpurified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

1.76 U/mg protein (on pNP-α-Man) at pH 7.0 and 37^oC.

One Unit of pNP-α-Man activity is defined as the amount of enzyme required to release 1 umol 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 BtMan92G on different substrates^a.

^aReactions were performed with 5 mM 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.

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: suggested use 7.0

Temperature Optima: suggested use 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.