Purification and Properties of the Acidic N-acetyl-&lt;I&gt;β&lt;/I&gt;-D-glucosaminidase from Porcine Semen*

Huang, Xiaohong; Luo, Zhen; Huang, Yi-Fan; Chen, Xinying; Huang, Junping

doi:10.3724/sp.j.1145.2009.00819

Cited by 1 publication

(3 citation statements)

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“…The isoelectric point of the enzyme was determined by IFGE. The result shows that the pI value of the enzyme was approximately 5.54, which is close to that of porcine semen acidic NAGase (Huang et al, 2009), and same as papaya NAGase (Chen et al, 2011). However, this value was different from that of the Bombyx mori alimentary canal (pI 4.86) (Koga et al, 1986) and Penaeus vannamei (pI 4.8) NAGases.…”

Section: Determination Of the Isoelectric Point Of The Enzymementioning

confidence: 58%

“…NAGase A and B are purified from bovine spleen, with NAGase B having only one subunit with a molecular weight of 140 kD (Vehpoorte, 1972). Pig semen alkaline NAGase has only one subunit, and its molecular weight is 58.03 kD (Huang et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

“…The E a was assayed by measuring the maximum velocity of the enzymatic hydrolysis of pNP-NAG, and the value of E a was calculated from the plot of log V m versus 1/T to be 64.19 kJ/mol, which is different for a variety of organisms. It has been reported that the E a values of NAGase from pig semen alkaline (Huang et al, 2008), pig semen acid (Huang et al, 2009), muscovy duck testicle (Wang et al, 2007), hetian chicken testicle and avian broilers testicle (Jin et al, 2009) are 88.73, 41.70, 69.05, 85.74 and 73.47 kJ/mol, respectively. The degree of catalytic reaction was as follows: pig semen alkaline > hetian chicken testicle > avian broilers testicle > muscovy duck testicle > bovine bull testicle > pig semen acid.…”

Section: Enzyme Activation Energymentioning

confidence: 99%

See 2 more Smart Citations

Purification and properties of N-acetyl--D glucosaminidase from bovine testicle

Huang¹,

Jin²,

Huang³

et al. 2013

Afr. J. Biotechnol.

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N-Acetyl-β-D-glucosaminidase (NAGase) from bovine testicle was purified by ammonium sulfate fractionation followed by diethylaminoethyl (DEAE)-cellulose (DEAE-32) and Sephacryl S-300 chromatography. The enzyme was purified to homogeneity as analyzed by polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing gel electrophoresis (IFGE). The specific activity of the purified enzyme was 658.21 U/mg. The enzyme was a single subunit with molecular weight of 68.3 kDa and contained 3.03% sugar. The pI value was calculated to be 5.54 using IFGE. The optimal pH and temperature of the enzyme for hydrolysis of p-Nitrophenyl-N-acetyl-β-D-glucosaminide (pNP-NAG) were found to be pH 5.6 and 50°C, respectively. The kinetics results showed that the enzyme hydrolyzed pNP-NAG following Michaelis-Menten with K m of 0.71 mM and V m of 16.72 M/min at pH 5.6 and 37°C. The enzyme was stable at pH values ranging from 2 to 6.5 and at temperatures below 60°C. The activation energy was determined to be 64.19 kJ/mol.

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Section: Determination Of the Isoelectric Point Of The Enzymementioning

confidence: 58%

Section: Discussionmentioning

confidence: 99%