Biochemical, Immunological and Physiological Studies on some Plant Pathogenic Fungi

Not availabl

Anticancer Activity of L-Asparaginase Produced from Vigna Unguiculata

The present study aimed to production of purified L-asparaginase from Vignaunguiculata. Different physiological parameters, such as pH, temperature and incubation period, were optimized for growth and maximum L-asparaginase production. The optimum parameters were 37°C, 30 min and pH 8.5. Maximum L-asparaginase was 886.4U/ml with a specific activity of 1140.7 U/ml (31 fold purification with 28 %yield) were obtained at optimum conditions. The purified L-asparaginase produced from Vignaunguiculata was used for characterization and general properties. The effect of pH and temperature on L-asparaginase activity as well as stability at different pH and temperature were determined. The optimum pH 8.5 and 37ºC temperature on L-asparaginase showed 100% residual activity. Stability of pH around 8.5 and temperature 70ºC showed 90 and 78 % residual activity at 30 and 60 min respectively. The L-asparaginase showed high stability at alkaline pH (pH 8.5) when incubated for up to 60h.The molecular weight of the produced L-asparaginase was close to 68.5 kDa. Cytotoxic activity of L-asparaginase was examined in vitro using four carcinoma cell lines. L-aspargenase has higher effective in growth inhibition against HEPG2 and HCT-116 but lower against HELLA and MCF7 carcinoma cell lines. The data show that L-aspargenase has a higher cytotoxic activity against HEPG2 and HCT116, revealed higher percentage of cell death, indicating antitumor properties, and demonstrate direct effect on cancer cell proliferation of HEPG2 and HCT116. Therefore, Vignaunguiculata was considered to be a suitable source for production of L-asparaginase has higher activity and good stability. Purified L-asparaginase obtained from Vignaunguiculata could be employed in drug chemotherapy and treatment of cancer

Biosorption technology for removal of metallic pollutants-An overview

The main sources of metallic pollutants to the environment are the diffuse sources such as forests and agricultural soils as well as industrial and municipal wastes, which are either discharged directly or transported in to the environment. Various conventional technologies such as chemical precipitation, solvent extraction, ion exchange, membrane separation, electrochemical treatment etc. have been employed to remove metal pollutants from aqueous solution. The exploration of new technologies involves the removal of toxic metals from wastewater with the use of biological adsorption technology. The biosorption is the selective appropriate process for removal of metal ions uptake that may involve the contribution of diffusion, adsorption, chelation, complexation, coordination, or micro-precipitation mechanisms, depending on the specific substrate (biomass). In this overview, the use of the various low cost, easily available and eco-friendly biosorbents used for removal of metallic pollutants from contaminated water and their mechanism are discussed

Studies on Phytohormone Producing Plant Pathogenic Fungi

Not availabl

Synthesis, detection and quantification of inulooligosaccharides and fructooligosaccharides by extracellular and intracellular inulinase and fructosyltransferase enzymes isolated from coprophilous fungi.

Masters Degree. University of KwaZulu-Natal, Durban.Exploration of fungal biodiversity capable of producing fructosyltransferase and inulinase enzymes in significant amounts is crucial for the production of oligofructans. Indigenous coprophilous fungi are predominantly sustainable bioresources, harbouring novel enzymes with potential industrial and biotechnological applications. Fructosyltransferase (Ftase) and inulinase are gaining considerable attention due to their capability to synthesise biofunctional nutraceuticals with low calories and health benefits when ingested in recommended dosages. Hence, due to several health benefits associated with prebiotics, bioprospecting for coprophilous fungi as unique bioresources of fructosyltransferase and inulinase was imperative. The present study therefore focused on the collection of herbivore dung from various terrestrial habitats in KwaZulu-Natal Province, South Africa whereby sixty-one (61) indigenous coprophilous fungal strains were isolated after repeated purification to monoculture. The axenic fungal strains were identified using morpho-taxonomic keys and molecular identification by 18S rDNA sequencing where Neocosmospora spp, Trichoderma spp., Aspergillus spp and Fusarium spp. were dominant. The fungal strains were subsequently assessed for their ability to produce extracellular and intracellular Ftase and inulinase enzymes. During the preliminary screening, the culture filtrate was examined for transfructosylating and hydrolytic activity using 2,3,5-triphenyl tetrazolium chloride (TTC) as a chromogenic marker and Lugol’s iodine solution, respectively. Zones of hydrolysis on 30 fungal isolates were observed on the TTC assay plates in diameters ranging from 15 mm to 30 mm, representing high extracellular Ftase activity. The formation of clear zones following addition of iodine solution on inulin rich media indicated the presence of inulinolytic activity. Secondary screening involved DNS assays of eight (8) isolates that secreted high concentrations of Ftase while six (6) different fungal strains showed <50 % inulinase: invertase ratio. The final screening step was tertiary screening where products of biocatalysis were qualitatively detected by thin layer chromatography to visualize saccharide spots of fructooligosaccharides and inulooligosaccharides. HPLC analysis of Ftase and inulinase reaction products revealed and further confirmed that coprophilous fungi harbour fructosyltransferase and inulinase enzymes. The crude extracellular fructosyltransferase enzyme was partially purified by 9.3-fold with a yield of 7.3 % and a specific activity of 2465.5 U mg-1 after a three-step procedure involving (NH4)2SO4 fractionation, dialysis and ion exchange chromatography. The apparent molecular weight of this Ftase was estimated by SDS-PAGE to be approximately 70 kDa. Zymogram analysis under non-reducing conditions showed the enzyme migrating as a polydisperse aggregate yielding broad band of approximately 100 kDa. The enzyme further exhibited an enhanced activity at a broad pH range of 4.0 – 8.0 and optimal activity at a temperature range of 40 °C – 80 °C, while the enzyme was stable at pH 8.0 and between 40 °C – 60 °C, respectively. Under these conditions, the enzyme remained stable retaining 95 % residual activity after incubation for 6 h. The presence of metal ions such as Hg2+ and Ag2+ inhibited Ftase activity while, Ca2+, Mg2+ and K+ at 1 mM increased the enzyme activity, with stabilization observed with Na+, Zn2+ and Cu2+. With sucrose as the substrate, the enzyme kinetics fitted the Michaelis-Menten model. The Km, Vmax and kcat values were 2.076 mM, 4.717 μmole min-1, and 4.7 min-1, respectively with a catalytic efficiency of 2.265 μmole min-1. In vitro antioxidant potential of FOS by 1,1 - diphenyl-2-picryl hydroxyl (DPPH) assay, ferric reducing antioxidant power (FRAP) assay and nitric oxide (NO) radical inhibition yielded IC50 of 6.71 μg/ml, 1.76 μg/ml and IC25 of 0.27 μg/ml, respectively. Free radical scavenging and inhibition activities showed a concentration-dependent antioxidant activity with no significant differences with oligosaccharide standards (p < 0.01). However, vitamin C was significant in FRAP and NO assays. These results clearly demonstrated that an indigenous coprophilous fungus is a potential new reservoir of salient biotechnological enzymes that can be exploited for the production of prebiotics for subsequent biotechnological applications

Statement complementing the EFSA Scientific Opinion on application (EFSA-GMO-UK-2006-34) for authorisation of food and feed containing, consisting of and produced from genetically modified maize 3272

Following a request from the European Commission, the GMO Panel assessed additional information related to the application for authorisation of food and feed containing, consisting of and produced from genetically modified (GM) maize 3272 (EFSA-GMO-UK-2006-34). The applicant conducted new agronomic, phenotypic and compositional analysis studies on maize 3272 and assessed the allergenic potential of AMY797E protein, addressing elements that remained inconclusive from previous EFSA opinion issued in 2013. The GMO Panel is of the opinion that the agronomic and phenotypic characteristics as well as forage and grain composition of maize 3272 do not give rise to food and feed safety, and nutritional concerns when compared to non-GM maize. Considering the scope of this application and the characteristics of the trait introduced in this GM maize, the effect of processing and potential safety implications of specific food or feed products remain to be further investigated. Regarding the allergenic potential of AMY797E protein and considering all possible food and feed uses of maize 3272, the Panel concludes that the information provided does not fully address the concerns previously raised by the Panel in 2013. Owing to the nature and the knowledge available on this protein family, it is still unclear whether under specific circumstances the alpha-amylase AMY797E has the capacity to sensitise certain individuals and to cause adverse effects. To further support the safety of specific products of maize 3272, the applicant provided thorough information relevant for the allergenicity assessment of dried distiller grains with solubles (DDGS), which is the main product of interest for importation into the EU. Having considered the information provided on this product, the Panel is of the opinion that under the specific conditions of use described by the applicant, DDGS produced from maize 3272 does not raise concerns when compared to DDGS from non-GM maize

Statement complementing the EFSA Scientific Opinion on application (EFSA‐GMO‐UK‐2006‐34) for authorisation of food and feed containing, consisting of and produced from genetically modified maize 3272

Following a request from the European Commission, the GMO Panel assessed additional information related to the application for authorisation of food and feed containing, consisting of and produced from genetically modified (GM) maize 3272 (EFSA‐GMO‐UK‐2006‐34). The applicant conducted new agronomic, phenotypic and compositional analysis studies on maize 3272 and assessed the allergenic potential of AMY797E protein, addressing elements that remained inconclusive from previous EFSA opinion issued in 2013. The GMO Panel is of the opinion that the agronomic and phenotypic characteristics as well as forage and grain composition of maize 3272 do not give rise to food and feed safety, and nutritional concerns when compared to non‐GM maize. Considering the scope of this application and the characteristics of the trait introduced in this GM maize, the effect of processing and potential safety implications of specific food or feed products remain to be further investigated. Regarding the allergenic potential of AMY797E protein and considering all possible food and feed uses of maize 3272, the Panel concludes that the information provided does not fully address the concerns previously raised by the Panel in 2013. Owing to the nature and the knowledge available on this protein family, it is still unclear whether under specific circumstances the alpha‐amylase AMY797E has the capacity to sensitise certain individuals and to cause adverse effects. To further support the safety of specific products of maize 3272, the applicant provided thorough information relevant for the allergenicity assessment of dried distiller grains with solubles (DDGS), which is the main product of interest for importation into the EU. Having considered the information provided on this product, the Panel is of the opinion that under the specific conditions of use described by the applicant, DDGS produced from maize 3272 does not raise concerns when compared to DDGS from non‐GM maize

Isolation and Screening of Glutaminase & Urease Free Novel Fungal Strains for the Production of L-Asparaginase

L - Asparaginase is an amidohydrolase that catalyzes the hydrolysis of amino acid L - aspar a gine in to aspartic acid and ammonia. It is used in the treatment of Acute L ymphoblastic Leukemia (ALL ) and some other malignant lymphoid abnormalities . It is also used in food industry to prevent the formation of a crylamide , a carcinogenic substance in carbohydrate rich fried and baked foods. Naturally L - Asparaginase is present i n plants, animals and microbes but microorganisms such as bacteria, yeast and fungi are generally used for the production of L - Asparaginase as it is difficult to obtain the same from plants and animals. It is found that the L - Asparaginase from bacteria causes side effects in humans including anaphylaxis and serious allergic reactions which can be fatal in some cases . To overcome this, eukaryotic organisms such as fungi can be used for the production of L - Asparaginase . Bu t sometimes the fungi produces L - glutaminase and urease enzymes along with L - Asparaginase which is difficult to remove in the purification stage. In order to prevent this fungal strains which can produce L - Asparaginase fre e of L - glutaminase and urease are isolated from different sources using standard protocols