Activity Staining and Inhibition Characterization of Dipeptidylpeptidase-III Enzyme from Goat Brain

Dipeptidylpeptidase-III (DPP-III) from goat brain was purified and characterized using Arginyl-Arginyl-4-methoxy-β-naphthylamide (Arg-Arg-4mβNA) substrate. This enzyme retained its activity in native 10% polyacrylamide gel when stained using Arg-Arg-4mβNA. The activity was significantly increased by 100 mM chloride. Studies for its inhibition with some peptides and chemical inhibitors revealed that Leu-Trp-Met-Arg-Phe-Ala was most potent inhibitor followed by Arg-Phe-Ala and Gly-Phe-Leu. All the studied chemical inhibitors caused 40–50% inhibition at 1 mM. Metal ions helped to regain activity of EDTA pretreated enzyme. ZnCl2 at 50 μM almost completely restored the enzyme activity. Further ZnCl2 and CoCl2 exerted protective effects on EDTA pretreated enzyme for its susceptibility to DTNB inhibition. Therefore, DPP-III is a metalloprotease with the involvement of cysteine residues either located at the catalytic site or involved in regulation.

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Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity

Environmental Toxicology and Pharmacology ◽

10.1016/j.etap.2016.04.011 ◽

2016 ◽

Vol 44 ◽

pp. 134-139 ◽

Cited By ~ 75

Author(s):

Murat Kucuk ◽

İlhami Gulcin

Keyword(s):

Enzyme Activity ◽

Carbonic Anhydrase ◽

Metal Ions ◽

Black Sea ◽

Salmo Trutta ◽

Purification And Characterization ◽

Sea Trout ◽

Carbonic Anhydrase Enzyme

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Purification and characterization of the carbonic anhydrase enzyme from horse mackerel (Trachurus trachurus) muscle and the impact of some metal ions and pesticides on enzyme activity

Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology ◽

10.1016/j.cbpc.2019.108605 ◽

2019 ◽

Vol 226 ◽

pp. 108605 ◽

Cited By ~ 13

Author(s):

Cuneyt Caglayan ◽

Parham Taslimi ◽

Cebrahil Türk ◽

Fatih Mehmet Kandemir ◽

Yeliz Demir ◽

...

Keyword(s):

Enzyme Activity ◽

Carbonic Anhydrase ◽

Metal Ions ◽

Horse Mackerel ◽

Trachurus Trachurus ◽

Purification And Characterization ◽

The Impact ◽

Carbonic Anhydrase Enzyme

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Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14

Marine Drugs ◽

10.3390/md17120681 ◽

2019 ◽

Vol 17 (12) ◽

pp. 681

Author(s):

Junhao Sun ◽

Xu Han ◽

Guanrui Song ◽

Qianhong Gong ◽

Wengong Yu

Keyword(s):

Enzyme Activity ◽

Metal Ions ◽

Animal Experiments ◽

Basic Research ◽

Facilitated Diffusion ◽

Divalent Metal Ions ◽

Polysaccharide Lyase ◽

Functional Studies ◽

Dose Dependent

Glycosaminoglycan (GAG) lyase is an effective tool for the structural and functional studies of glycosaminoglycans and preparation of functional oligosaccharides. A new GAG lyase from Microbacterium sp. H14 was cloned, expressed, purified, and characterized, with a molecular weight of approximately 85.9 kDa. The deduced lyase HCLaseM belonged to the polysaccharide lyase (PL) family 8. Based on the phylogenetic tree, HCLaseM could not be classified into the existing three subfamilies of this family. HCLaseM showed almost the same enzyme activity towards hyaluronan (HA), chondroitin sulfate A (CS-A), CS-B, CS-C, and CS-D, which was different from reported GAG lyases. HCLaseM exhibited the highest activities to both HA and CS-A at its optimal temperature (35 °C) and pH (pH 7.0). HCLaseM was stable in the range of pH 5.0–8.0 and temperature below 30 °C. The enzyme activity was independent of divalent metal ions and was not obviously affected by most metal ions. HCLaseM is an endo-type enzyme yielding unsaturated disaccharides as the end products. The facilitated diffusion effect of HCLaseM is dose-dependent in animal experiments. These properties make it a candidate for further basic research and application.

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Occurrence of an endo-1,3-β-glucanase in culture fluids of the yeast Candida utilis. Purification and characterization of the enzyme activity

Biochemical Journal ◽

10.1042/bj1770107 ◽

1979 ◽

Vol 177 (1) ◽

pp. 107-114 ◽

Cited By ~ 16

Author(s):

T G Villa ◽

V Notario ◽

J R Villanueva

Keyword(s):

Enzyme Activity ◽

Gel Electrophoresis ◽

Candida Utilis ◽

Culture Medium ◽

Polyacrylamide Gel Electrophoresis ◽

Gel Filtration ◽

Polyacrylamide Gel ◽

Beta Glucanase ◽

Hydrolysis Of

The endo-1,3-beta-glucanase (EC 3.2.1.6) secreted into the culture medium by cells of Candida utilis was isolated and purified to homogeneity on polyacrylamide-gel electrophoresis and in ultracentrifugation studies (s20,w = 1.97S). The purified enzyme represented only 0.001% of the total 1,3-beta-glucanase activity, the remainder being due to an exo-1,3-beta-glucanase enzyme, and behaved as an acidic glycoprotein (pI 3.3) in isoelectric-focusing experiments. The mol.wt. was estimated to be 21 000 by gel filtration and polyacrylamide-gel electrophoresis. Studies on the hydrolysis of different substrates showed that the enzyme was only able to break down (1 leads to 3)-beta-linkages, by an endo-splitting mechanism. Glucono-delta-lactone, D-glucoronolactone and heavy metal ions such as Hg2+ were inhibitors of the enzyme activity. The function of this endo-beta-glucanase in C. utilis is discussed.

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