PRELIMINARY STUDIES ON ISOLATION, PURIFICATION AND SOME PROPERTIES OF THE β-N-ACETYL-D-GLUCOSAMINIDASE FROM ERIOCHEIR SINENSIS

HUANG Xiao-Hong; CHEN Hong-Hui; HUANG Yi-Fan

Volume 31 Issue 4

Aug. 2014

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ACTA HYDROBIOLOGICA SINICA > 2007 > 31(4): 563-569.

HUANG Xiao-Hong, CHEN Hong-Hui, HUANG Yi-Fan. PRELIMINARY STUDIES ON ISOLATION, PURIFICATION AND SOME PROPERTIES OF THE β-N-ACETYL-D-GLUCOSAMINIDASE FROM ERIOCHEIR SINENSIS[J]. ACTA HYDROBIOLOGICA SINICA, 2007, 31(4): 563-569.

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PRELIMINARY STUDIES ON ISOLATION, PURIFICATION AND SOME PROPERTIES OF THE β-N-ACETYL-D-GLUCOSAMINIDASE FROM ERIOCHEIR SINENSIS

Institute of Animal Health, College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002

Received Date: June 20, 2006
Rev Recd Date: March 16, 2007
Published Date: July 24, 2007

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Abstract

Abstract

A β-N-Acetyl-D-glucosaminidase (EC3.2.1.30) was purified from the viscera of Edocheir sinensis by ammonium sulfate fractionation,chromatography on DEAE-32, Sephadex G-100 and DEAE-32. The purified enzyme preparation was homogeneous as judged by polyacrylamide gel electrophoresis. The specific activity of the enzyme was 4490.79U/rag. The molecular weight of the subunits was determined to be 121.21,98.63 and 73.48kD,respectively. The pI value was calculated to be 4.5 by isoelectric focusing. The optimum temperature and pH of the enzyme for the hydrolysis of p-nitrophenyl-N-acetyl-β-D-glucosaminide (pNP-NAG) were determined to be at 45℃and at pHS.5,respectively. The enzyme was stable in the pH ranges of 4.9 to 9.3 under 37℃ and at temperatures below 40℃. The enzyme follows typical Michaelis-Menten kinetics for the hydrolysis of pNP-β-D-GlcNAc.The Km and Vm values were determined to he 0.357mmo/L and 10.41μmol/L min at pH5.6 and 37℃, respectively.The activation energy of the enzyme for the hydrolysis of pNP-β-D-GlcNAc was to be 76.50kJ/mol.The effects of metal ions on the enzyme were studied. Mg2+, Ca2+ and Ba2+ had not influenced the enzyme activity. Na+ activated the enzyme, while, Li+, K+, Zn2+, Hg2+, Pb2+, Cu2+, and Al3+ showed various degrees of inhibitory effects on the enzyme.
- Eriocheir sinensis,
- B-N-Acety-l D-glucosaminidase,
- Isolation and purification,
- Kinetics,
- Stability

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PRELIMINARY STUDIES ON ISOLATION, PURIFICATION AND SOME PROPERTIES OF THE β-N-ACETYL-D-GLUCOSAMINIDASE FROM ERIOCHEIR SINENSIS

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