CcSia

Ex-Sia0214

(EC. 3.2.1.18) exo-Sialidase

CAZy Family: GH33

PROPERTIES

1. ELECTROPHORETIC PURITY

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

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

2. SPECIFIC ACTIVITY

38.5 U/mg protein (on 4-MU-NeuAc) at pH 7.0 and 37°C.

One unit of enzyme was defined as the amount of enzyme that generate 1 μmol of 4-MU per min..

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Figure 2. HPLC analysis of the transformed product from gangliosides mixture by recombinant sialidase. Blue dot represents the gangliosides substrate; black solid line represents the transformed product.

Table1 Biotransformation efficiency of gangliosides by recombinant sialidase CcSia.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 5.0

pH Stability: 4.0-11.0

Temperature Optima: 45°C

5. STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in acetate buffer (20 mM) pH 4.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Yuan Y, Zhou YF, et al. High yield preparation of ganglioside GM1 using recombinant sialidase from Cellulosimicrobium cellulans. Process Biochemistry. 2017,58: 92-97.

[2] Ji L, Yuan Y, et al.Preparation of Ganglioside GM1 by Supercritical CO2 Extraction and Immobilized Sialidase. Molecules. 2019,24,3732.