BsGal42A（endo-β-1,4-Galactanase）

BsGal42A

Endo-Gal0058

(EC.3.2.1.89)endo-β-1,4-Galactanase

CAZy Family: GH42


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~74 kDa)

Figure 1. Electrophoresis analysis of BsGal42A. 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, BsGal42A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

0.083 U/mg protein (on pNP-β-gal) at pH 6.5 and 40°C.

One Unit of pNP-β-gal activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-β-gal (4 mM) in ammonium acetate (50 mM) pH 6.5.


3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BsGal42A on different substrates.


4.PHYSICOCHEMICAL PROPERTIES

pH Optima:6.5

pH Stability: 6.0-8.0

Temperature Optima: 40°C

Temperature Stability: <45°C

Figure 2. Biochemical charcterization of BsGal42A. (A) Effect of temperature on the BsGal42A activity carried out at pH 6.5 and 8 mM ofpNP-βGal, (B) Effect of pH on the BsGal42A activity using McIlvaine buffers (pH 5-8) at 40 °C and 8 mM of pNP-βGal. Data points are the average of three independent sets of experiments and the error bars represent the mean ± SD. All assays were performed in triplicate.




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]Lara A, Li Y, Paul D, et al.Characterization of a β-galactosidase from Bacillus subtilis with transgalactosylation activity.International Journal of Biological Macromolecules. 2018,120:279-287.

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BsGal42A（endo-β-1,4-Galactanase）

BsGal42A

Endo-Gal0058

(EC.3.2.1.89)endo-β-1,4-Galactanase

CAZy Family: GH42


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~74 kDa)

Figure 1. Electrophoresis analysis of BsGal42A. 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, BsGal42A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

0.083 U/mg protein (on pNP-β-gal) at pH 6.5 and 40°C.

One Unit of pNP-β-gal activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-β-gal (4 mM) in ammonium acetate (50 mM) pH 6.5.


3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BsGal42A on different substrates.


4.PHYSICOCHEMICAL PROPERTIES

pH Optima:6.5

pH Stability: 6.0-8.0

Temperature Optima: 40°C

Temperature Stability: <45°C

Figure 2. Biochemical charcterization of BsGal42A. (A) Effect of temperature on the BsGal42A activity carried out at pH 6.5 and 8 mM ofpNP-βGal, (B) Effect of pH on the BsGal42A activity using McIlvaine buffers (pH 5-8) at 40 °C and 8 mM of pNP-βGal. Data points are the average of three independent sets of experiments and the error bars represent the mean ± SD. All assays were performed in triplicate.




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]Lara A, Li Y, Paul D, et al.Characterization of a β-galactosidase from Bacillus subtilis with transgalactosylation activity.International Journal of Biological Macromolecules. 2018,120:279-287.