BtMan130A

Ex-Man0113

(EC.3.2.1.197)exo-β-1,2-Mannosidase

CAZy Family: GH130

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~43 kDa)

Figure 1. Electrophoresis analysis ofBtMan130A. 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, BtMan130A purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtMan130A on different substrates.

^aFor substrates where the two enzymes displayed no activity, assays were carried out in both 20 mM Na-Hepes

and 50 mM sodium phosphate buffers, pH 7.5. and 50 mM sodium phosphate buffers, pH 7.5.

Figure.2 HPAEC and TLC analysis of BtMan130A catalyzed reactions.BtMan130A was incubated with 1mg/mlC. albicans mannan for up to 16 h. Samples were removed at regular intervals and analysed by TLC and HPAEC

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: suggested use 7.5

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, Baslé A, Ladevèze S, et al. The GH130 family of mannoside phosphorylases contains glycoside hydrolases that target β-1, 2-mannosidic linkages in Candida mannan. Journal of Biological Chemistry, 2015, 290(41): 25023-25033.