TmGH28A

Exo-Gal0206

(EC.3.2.1.82) exo-α-Galacturonosidase

CAZy Family: GH28

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~50 kDa)

Figure 1. Electrophoresis analysis of TmGH28A. 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, TmGH28A purified from Ni sepharose fastflow column.

2. SPECIFIC ACTIVITY

216 U/mg protein (on oligogal-acturonic acid) at pH 6.5 and 80°C.

One Unit of activity is defined as the amount of enzyme required to release 1 μmol of reducing end groups per minute from oligogalacturonic acid (12 mM) in phosphate buffer (100 mM) pH 6.5.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Kinetic parameters of TmGH28A fromT. maritimaon saturatedoligogalacturonates (GalpA) with length n= 2–8.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 6.7

Temperature Optima: 80°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. REFERENCE

[1] Leon D. Kluskens, Gert-Jan W.M. van Alebeek, et al. Characterization and mode of action of an exopolygalacturonase from the hyperthermophilic bacterium Thermotoga maritima. FEBS Lett. 2005 ,272(21) :5464-73.