BtMan76C

Endo-Man0101

(EC.3.2.1.101)endo-α-1,6-Mananase

CAZy Family: GH76

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~45 kDa)

Figure 1. Electrophoresis analysis of BtMan76C. 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, BtMan76C purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Figure.2 a, b, BT3792 and BtMan76C, respectively, were incubated with α-1,6-mannotetraose. 

Figure.3 BT3792 and BtMan76C were incubated with unbranched yeast mannan. The yeast mannan at 0.1% was incubated with the two GH76 α-1,6-mannanases for 1 h at 37 ℃ in 50 mM sodium phosphate buffer, pH 7.0. The limit products were analysed by TLC. α-1,6-Mannooligosaccharides are identified by their degree of polymerization (M1, mannose; M2, mannobiose; M3, mannotriose; M4, mannotetraose).

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.