Purification and characterization of Se‐enriched Grifola frondosa glycoprotein, and evaluating its amelioration effect on As 3+ ‐induced immune toxicity

Using commercial API-ZYM screening kits, highly active α-glucosidase, β-glucosidase, and β-N-acetylglucosaminidase were found in Grifola frondosa, having potential for carbohydrate utilization. Of these, β-N-acetylglucosaminidase, which converts chitin to N-acetylglucosamine, was purified and characterized. The recovery was 24.5%, and the purified enzyme had a specific activity 0.67 U/mg protein. Chitinase activity was confirmed by zymogram analysis. The enzyme was also shown to be β-N-acetylglucosaminidase, as N-acetylglucosamine was the main hydrolysis product from colloidal chitin. Thus, the molecule was named NAG38, to indicate β-N-acetylglucosaminidase activity and a molecular weight of 38 kDa, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Enzymatic activity was optimal at pH 7.0 and 50 °C, with Km and Vmax values of 0.112 mM and 0.570 μmol/min/mg protein against p-nitrophenyl N-acetyl-β-D-glucosaminide. The bioactivity was inhibited by Hg2+, Ag+, Mg2+, Zn2+, Ca2+, and Mn2+, with residual enzyme bioactivity only 11.1% after incubation in Hg2+, but was not substantially inhibited by Ba2+, K+, and Na+.

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Cloning, expression, purification and characterization of HSP105

Cell Biology International ◽

10.1042/cbi034s046a ◽

2010 ◽

Vol 34 (8) ◽

pp. S46-S46

Author(s):

Dong Han ◽

Huang Xu

Keyword(s):

Purification And Characterization

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Cloning and high level expression of Cynodon dactylon (Bermuda grass) pollen profilin (Cyn d 12) in Escherichia coli: purification and characterization of the allergen

Clinical & Experimental Allergy ◽

10.1046/j.1365-2222.1997.1470952.x ◽

1997 ◽

Vol 27 (11) ◽

pp. 1307-1313 ◽

Cited By ~ 2

Author(s):

J. A. ASTURIAS ◽

M. C. ARILLA ◽

N. GOMEZ-BAYON ◽

J. MARTINEZ ◽

A. MARTINEZ ◽

...

Keyword(s):

Escherichia Coli ◽

Grass Pollen ◽

Cynodon Dactylon ◽

Bermuda Grass ◽

Purification And Characterization ◽

High Level Expression ◽

Bermuda Grass Pollen ◽

D 12 ◽

High Level

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Purification and characterization of a (1 → 3)-β-?-glucan-binding protein from horseshoe crab (Tachypleus tridentatus) amoebocytes

Carbohydrate Research ◽

10.1016/s0008-6215(96)00214-5 ◽

1996 ◽

Vol 295 (1-4) ◽

pp. 103-116

Author(s):

H Tamura

Keyword(s):

Horseshoe Crab ◽

Binding Protein ◽

Purification And Characterization ◽

Tachypleus Tridentatus

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Purification and characterization of phosphoglucomutase from peas

Physiologia Plantarum ◽

10.1034/j.1399-3054.1994.920317.x ◽

1994 ◽

Vol 92 (3) ◽

pp. 479-486 ◽

Cited By ~ 1

Author(s):

Cynthia M. Galloway ◽

W. Mack Dugger

Keyword(s):

Purification And Characterization

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Purification and Characterization of Tissue-Type Plasminogen Activator in Maxillary Mucosa with Chronic Inflammation

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657879 ◽

1985 ◽

Vol 54 (02) ◽

pp. 485-489 ◽

Cited By ~ 3

Author(s):

Yukiyoshi Hamaguchi ◽

Masuichi Ohi ◽

Yasuo Sakakura ◽

Yasuro Miyoshi

Keyword(s):

Plasminogen Activator ◽

Chronic Inflammation ◽

Gel Filtration ◽

Potassium Acetate ◽

Tissue Type ◽

Purification And Characterization ◽

Tissue Type Plasminogen Activator ◽

Urokinase Inhibitor ◽

Type Plasminogen Activator

SummaryTissue-type plasminogen activator (TPA) was purified from maxillary mucosa with chronic inflammation and compared with urokinase. Purification procedure consisted of the extraction from delipidated mucosa with 0.3M potassium acetate buffer (pH 4.2), 66% saturation of ammonium sulfate, zinc chelate-Sepharose, concanavalin A-Sepharose and Sephadex G-100 gel filtration chromatographies.The molecular weight of the TPA was approximately 58,000 ± 3,000. Its activity was enhanced in the presence of fibrin and was quenched by placental urokinase inhibitor, but not quenched by anti-urokinase antibody. The TPA made no precipitin line against anti-urokinase antibody, while urokinase did.All these findings indicate that the TPA in maxillary mucosa with chronic inflammation is immunologically dissimilar to urokinase and in its affinity for fibrin.

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