BtGal35B

Exo-Gal0069

(EC.3.2.1.23) exo-β-1,6-galactosidase

CAZy Family: GH35

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~89kDa)

Figure 1. Electrophoresis analysis of BtGal35B. 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, BtGal35B purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

1.25 U/mg protein (on pNP-β-gal) at pH 7.0 and 37°C.

One Unit ofpNP-β-gal activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-β-gal (5 mM) in phosphate buffer (20 mM) pH 7.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtGal35B on different substrates^a.

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

^bAbsorption caused by releasedp-nitrophenol was measured at 405 nm. The relative activity on pNPβGal was taken as 100%.

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

Figure2. The structure of arabinogalactans, PULs upregulated by the glycans and enzymes that attack these glycans.

4.PHYSICOCHEMICAL PROPERTIES

pH Optima:suggesteduse7.0

Temperature Optima:suggesteduse37°C

5.STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in phosphate buffer (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Cartmell A, Muñoz-Muñoz J, Briggs JA, et al. A surface endogalactanase in Bacteroides thetaiotaomicron confers keystone status for arabinogalactan degradation. Nat Microbiol. 2018, 3(11): 1314-1326.