BsAra43A

Endo-Ara0136

(EC.3.2.1.99)endo-α-1,5-L-arabinanase

CAZy Family: GH43

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~53kDa)

Figure 1. Electrophoresis analysis of BsAra43A. 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, BsAra51B purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

72.8 U/mg protein (on Linear α-1,5-L-arabinan) at 37°C, pH 6.6.

One Unit of on Linear α-1,5-L-arabinanf activity is defined as the amount of enzyme required to release 1 mmol of reducing sugarper minute from Linear α-1,5-L-arabinan (0.2% (w/v)) in 200 mM phosphate/100 mM citrate buffer (PC buffer) pH 6.6.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Activity ofBsAra43Aagainst arabinose-containing substrates and biochemical properties

^aActivity was assayed by incubating purified recombinantBsAra43Awith substrates at 37°C, pH 6.6. The data shown are the averages ± standard deviations of three independent experiments, each performed in duplicate measurements.

^bNA, no detectable activity.

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 6.6

Temperature Optima: 50°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]Inácio J M, de Sá-Nogueira I. Characterization of abn2 (yxiA), encoding a Bacillus subtilis GH43 arabinanase, Abn2, and its role in arabino-polysaccharide degradation[J]. Journal of bacteriology, 2008, 190(12): 4272-4280.

[2]de Sanctis D, Inácio J M, Lindley P F, et al. New evidence for the role of calcium in the glycosidase reaction of GH43 arabinanases[J]. The FEBS Journal, 2010, 277(21): 4562-4574.