PpHgl9A (PpPel9a)

Ex-Hgl0310

(EC. 4.2.2.2) exo-Pectate lyase

CAZy Family: PL9

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~45 kDa)

Figure 1. Molecular weight of PpPel9a determined by SDS-PAGE: (M) protein molecular weight markers (PageRuler Prestained Protein Ladder, Thermo Scientific) (1) supernatant of cell lysis from E. coli BL21 (DE3) cells; (2), supernatant of cell lysis from recombinant E. coli BL21 (DE3) cells harboring pET-28a-pppel9a plasmid; (3) recombinant enzyme PpPel9a purified from Ni-NTA agarose column.

2. SPECIFIC ACTIVITY

298.5 U/mg protein (on PGA) at pH 10.0 and 37°C.

One unit of pectate lyase activity was defined as the enzyme needed to produce 1 umol of unsaturated bonds per min.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Degradation of various pectins by PpPel9a.

Figure 2. Analysis of the oligomers released from PGA by ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS). The oligosaccharides separated by UPLC was further analyzed by ESI-MS analysis (small box).

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 10.0

pH Stability: 6.0-11.0

Temperature Optima: 40°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. A Novel PL9 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression, and Its Application in Pectin Degradation. International Journal of Molecular Sciences. 2019, 20, 3060.