CjAra43A (CjAra2A)

Ex-Ara0129

(EC.3.2.1)exo-α-1,2-Arabinofuranosidase

CAZy Family: GH43

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~34 kDa)

Figure 1. Electrophoresis analysis ofCjAra43A. 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,CjAra43Apurified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

2.66 U/mg protein (on pNP-α-Araf) at pH 7.0and 37^oC

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

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity ofCjAra43Aon 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 pNPαAraf was taken as 100%.

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

Table 2.Kinetic properties of CjAra43A.

The enzymes were assayed in 50 mM sodium phosphate buffer, pH 7.0, at 25 and 37°C.

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 25°C

5.STORAGE CONDITIONS

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

6. REFERENCES

[1] Cartmell A, McKee L S, Peña M J, et al. The structure and function of an arabinan-specific α-1, 2-arabinofuranosidase identified from screening the activities of bacterial GH43 glycoside hydrolases[J]. Journal of Biological Chemistry, 2011, 286(17): 15483-15495.