Production of pectin lyase from xanthomonas campestris, purification, characterization and fruit juice application

Extracellular pectin lyase (EC 4.2.2.10) was produced by Xanthomonas campestris in solid state fermentation. Solid state fermentation was carried out using 250 mL Erlenmeyer flasks containing 5 g wheat bran, 1% citrus pectin and 10 mL salt mixture that composed of 0.14% (NH4)2SO4 0.02% MgCl2 and 0.02% K2HPO4. Pectin lyase enzyme was purified 43 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 77.5 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was controlled by SDS-PAGE. The optimum pH and temperature of enzyme was determined as pH 9.0 and 50°C, respectively. Pectin lyase was mostly stable 40°C for 24 hours. KM and Vmax were calculated respectively as 0.69 mg/mL and 3.84 |?mol/L?min. Purified pectin lyase was inhibited by Fe+, Zn2+, Ca2+, Co2+ and Hg2+ except for Cu2+ and Mg2+ The purified pectin lyase enzyme was used for getting fruits juices. It was determined that yields of fruits juices improved when it was compared with control

The synthesis, characterization, antimicrobial and antimutagenic activities of hydroxyphenylimino ligands and their metal complexes of usnic acid isolated from Usnea longissima

PubMed ID: 24589530Novel multifunctional hydroxyphenylimino ligands (L1, L2 and L3) were synthesized by the condensation of 2-aminophenol, 3-aminophenol and 4-aminophenol with usnic acid, a lichen metabolite. The synthesized ligands and their Cu(ii), Co(ii), Ni(ii) and Mn(ii) complexes were characterized using FT-IR, UV-Vis, 1H-NMR, 13C-NMR, 1D- and 2D NMR (DEPT, COSY, HMQC and HMBC), LC-MS and TGA. In addition, the metal complexes of the novel ligands were prepared with high yields using Cu(ii), Co(ii), Ni(ii) and Mn(ii) salts and were characterized using the FT-MIR/FAR, UV-Vis, elemental analysis, ICP-OES and TG/DTA techniques. The ligands and their complexes were tested against ten important pathogen microorganisms using the disc diffusion method and the metal complexes of the ligands were more active against all of the microorganisms tested with a broad spectrum than the ligands exhibiting 11–32 mm inhibition zones. On the other hand, a broad spectrum of the strongest antimicrobial activity was determined for the Mn(ii) and Cu(ii) complexes of the hydroxyphenylimino ligand with usnic acid (L3). In addition, the antimutagenic activities of all of the ligands and their metal complexes were determined using the Ames-Salmonella and E. coli WP2 microbial assay systems and they showed varied and strong antimutagenic effects. In general, it has been found that the Co and Mn complexes of the ligands possess potent antimutagenic activity. In view of these results, it can be concluded that some metal complexes can be used as antimicrobial and anticancer agents. © 2014 The Partner Organisations

Production of a novel pectin lyase from Bacillus pumilus (P9), Purification and characterisation and fruit juice application

Extracellular pectin lyase (EC 4.2.2.10) was produced by Bacillus pumilus (P9) in solid state fermentation. Pectin lyase enzyme was purified 36.36 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 25 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was controlled by SDS-PAGE. The optimum pH and temperature of enzyme was determined as pH 6.0 and 60oC, respectively. Pectin lyase was mostly stable at 40°C. Its' activity deceased in 50% for 1h at 60°C and 40% for 4 h at 50°C. Vmax and KM were calculated respectively as 0.298 mg/mL and 132.6 mol/L*min. The presence of 10 mM concentration of Ca2+, Cu2+, Mn2+, Mg2+, Zn2+, Hg2+, Fe2+ and EDTA, L-cystein, ascorbic acid and -mercaptoethanol significantly enhanced the pectin lyase of the purified enzyme. The purified pectin lyase enzyme was used for getting fruits juices. It was determined that yields of fruits juices significantly improved when it was compared with control. © 2010 University of Bucharest

Protective effects of methanol extracts from Cladonia rangiformis and Umbilicaria vellea against known mutagens sodium azide and 9-aminoacridine

PubMed ID: 21427134Lichens and their various extracts have been occasionally used in the treatment of many diseases. Cladonia rangiformis and Umbilicaria vellea are two important species of these lichens and they have several biological activities. In the present study, methanol extracts of these lichens, which are grown in the Eastern Anatolia Region of Turkey, were isolated, and their mutagenic and antimutagenic properties were investigated by using AMES-Salmonella and Zea mays Root Tip Mitotic Index mutagenicity and antimutagenicity assay systems. Known mutagens sodium azide (NaN3) and 9-Aminoacridine (9-AA) were used to determine antimutagenic properties of methanol extracts. The results showed that all methanol extracts, investigated in the present study, can be considered genotoxically safe because they do not have mutagenic activity at the tested concentrations. Besides, all of them have antimutagenic activity against 9-AA known as a model intercalator agent in the AMES-Salmonella test system. The inhibition rates obtained from the antimutagenicity assays ranged from 37.07% (C. rangiformis—5 µg/plate) to 54.39% (C. rangiformis—5 µg/plate). Furthermore, all the methanol extracts have significant antimutagenic activity against NaN3mutagenicity in Z. mays Root Tip Mitotic Index assay system. These activities are valuable towards an extension of the employ of these drugs as new phytotherapeutic or preservative ingredients. © 2011, SAGE Publications. All rights reserved