PoAra43C

Ex-Ara00345

(EC.3.2.1.99) endo-α-1,5-Arabinanase

CAZy Family: GH43

PROPERTIES

1. ELECTROPHORETIC PURITY

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

Figure 1. Electrophoresis analysis of PoAra43C. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate before Methanol induction; lane 2-4, culture lysat after Methanol induction; lane 5, PoAra43C purified from Ni sepharose fastflow column.

2. SPECIFIC ACTIVITY

2.71 U/mg protein (on Sugar Beet Linear Arabinan)at pH 5.0 and 50°C ;

0.04 U/mg protein (on Sugar Beet Arabinan) at pH 5.0 and 50°C;

One Unit of arabinanase activity is defined as the amount of enzyme required to release 1 μmol of reducing sugar per minute from Sugar Beet Linear Arabinan(2 mg/ml) in Na-Acetate buffer (20 mM) pH 5.0.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Figure 2. Release of reducing sugar from arabinan substrates by PoAra43C.

Figure 3. HPAEC analyses of the enzymatic hydrolysis products generated from arabino-oligosaccharides and Sugar Beet Linear Arabinan by PoAra43C.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 5.0

pH Stability: 5.0-6.0

Temperature Optima: 50°C

Temperature Stability: 30°C-40°C

Figure 4. Effect of pH on activity (a) and stability (b) of PoAra43C using Sugar Beet Linear Arabinan as substrate. The optimal pH (a) was determined at different pH from 2 to 11. The maximum activity obtained was defined as 100% activity. Thermal stability was determined by incubating the enzyme for 3 h at different pH. The activity of the enzyme before incubation was defined as 100%. Results are presented as means ± standard deviations(n = 3).

Figure 5. Effect of temperature on activity (a) and stability (b) of PoAra43C using Sugar Beet Linear Arabinan as substrate. The optimal temperature (a) was determined at different temperatures from 20 to 90℃. The maximum activity obtained was defined as 100% activity. Thermal stability was determined by incubating the enzyme for 3 h at different temperatures. The activity of the enzyme before incubation was defined as 100%. Results are presented as means ± standard deviations(n = 3).

5. STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in acetate buffer (20 mM) pH 5.0. Swirl to mix the enzyme immediately prior to use.