Purification and Characterization of Pyrophosphatase from Dorsal Muscle in Silver Carp (Hypophthalmichthys molitrix)

doi:10.7506/spkx1002-6630-201613023

Abstract

Abstract:

Pyrophosphatase (PPase) was extracted and purified from silver carp (Hypophthalmichthys molitrix) dorsal
muscle by crude extraction, 50%–80% saturated ammonium sulfate precipitation and DE-52 anion exchange column
chromatography. The purified enzyme migrated as a single band with a molecular mass of 40 kD on SDS-polyacrylamide
gel electrophoresis (SDS-PAGE). The enzymatic characterization showed that the purified enzyme could hydrolyze
pyrophosphates, and the time range for its initial velocity was 0–15 min. The optimum temperature for the purified PPase
was 45 ℃ and the optimum pH was 7.5. Mg2+ was necessary for PPase activation and the activity reached up to the
maximum value at Mg2+concentration of 5 mmol/L. The activity of PPase was strongly inhibited by 5 mmol/L Ca2+, Zn2+,
EDTA-Na2, EDTA-Na4, KIO3 or 1 mmol/L NaF. The kinetic constants vmax and Km of the purified PPase for tetrasodium
pyrophosphate as substrate were 0.051 U/mg and 0.54 mmol/L, respectively.

Key words: pyrophosphatase, purification, enzymatic characterization, dorsal muscle, silver carp

CLC Number:

TS201.2

LIU Wei, XU Meng, ZHANG Yawei, ZHOU Li, MA Zhifang, WANG Fulong, PENG Zengqi. Purification and Characterization of Pyrophosphatase from Dorsal Muscle in Silver Carp (Hypophthalmichthys molitrix)[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201613023.

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