Isolation, purification and characterization of inulin and fructooligosaccharides from chicorium intybus and inulinase from aspergillus niger

Inulin is a non-digestible carbohydrate fructan polymer consisting mainly of β (1→2) fructosyl fructose links. Enzymatic hydrolysis of inulin by inulinase results in the production of low D.P (degree of polymerization) oligosaccharides also called fructooligosaccharides(FOS). Isolation of inulin from chicory root (Chicorium intybus) was achieved by first, extraction using deionized water (600C), followed by carbonation (0.1 M Ca(OH)2 and CO2 gas). This was filtered in order to remove the non sugars, thereafter, treated successfully with polyamide 6 powder. A cation exchanger and an anion exchanger were used to further exclude other components such as tannins and pigments. The extracted inulin was quantified using the Somogyi-Nelson colourimetric assay. Chicory root (207 g, 30 % being water) yielded 30 g of the raw extract. A 100 mg of the raw extract was assayed and found to contain 11 % yield of inulin which was 80.2 % in purity and 4 % free fructose. Analysis of the crude and purified inulin extracts on the MALDI TOF spectrometry showed the samples to have a DP of 2 to 22 and 2 to 27 respectively. Maximum inulinase production from Aspergillus niger grown on inulin was observed after 60 hours. The enzyme activity was found to be 1.168 U/ml with a temperature and pH optimum of 30 °C and 7.7 respectively. The enzyme proved to be unstable as it progressively lost its total activity during attempts at purification

Gold mine tailings : a potential source of silica sand for glass making

Abstract:Mining of minerals such as gold, copper, and platinum has been one of several activities sustaining the economy of South Africa. However, the mining sector has contributed significantly to environmental contamination through the improper disposal of mine tailings which covers vast areas of land. Therefore, this study utilised a vitrification process to manufacture glass from gold mine tailings. X-ray fluorescence was used to determine the chemical composition of the tailings while X-ray diffraction was adopted for the mineralogy. The tailings were of granitic composition enriched in potentially toxic elements such as copper, cadmium, zinc, lead, arsenic, and chromium. A representative sample of gold mine wastes of sandy grain size was used in making the glass. Based on composition, the glass was formulated by adding an average 10.0 mass% of CaCO3 and 5.0mass%ofNa2CO3 to35.0mass%ofSiO2,whichresultedintheproductionofagreen-colouredglass

Mobility of Trace Element Contaminants from Abandoned Gold Mine Dump to Stream Waters in an Agricultural Active Area

In this study, the selected streams within the Blesbokspruit located in South Africa were characterised in this study. Because of prolonged mining activities coupled with ineffective management practices, several mine tailing (MT) dumps are widely distributed in this area. Metals and metalloid contamination from these tailing facilities have been reported to be major contributors to environmental hazards such as acid mine drainage (AMD). With increased agricultural activities in this area, an assessment of the general quality of water being utilised for irrigation purposes and feeding of farm animals becomes inevitable. A procedural method was implemented in a bid to identify relations between tailing and stream water contamination. Representative gold tailing sediments and water samples were collected, respectively. With the aid of X-ray fluorescence (XRF) and X-ray diffraction (XRD), the mineralogical characterisation of the tailing sediments was successfully carried out, while acid digestion using inductively coupled plasma-optical emission spectrometry (ICP-OES) was utilised in the determination of trace metal contents. Samples of different water sources were also characterised. There was a clear description of the link between tailings, water contamination and possible implications to animals and humans in the long run

Evaluation of Trace Elemental Levels as Pollution Indicators in an Abandoned Gold Mine Dump in Ekurhuleni Area, South Africa

In the Blesbokspruit area of Ekuhurleni, South Africa, previous gold mining activities resulted in many tailings dump sites. 20 representative soil samples were used in describing the distribution of metals. The soils were very strongly acidic ranging from 3.86 to 4.34 with a low cation exchange capacity (CEC). Based on X-ray fluorescence (XRF) analysis, elemental composition of the soils revealed average values of major elements such as Na2O (0.18%), MgO (0.63%), Al2O3 (6.51%), SiO2 (81.83%), P2O5 (0.04%), SO3 (3.40%), K2O (1.98%), CaO (0.45%), TiO2 (0.51%), Cr2O3 (0.17%), MnO (0.04%), Fe2O3 (3.59%), NiO (0.04%), As2O3 (0.02%), with Rb2O and SrO falling below 0.01%. Trace metals (TM) contamination levels in the soils were evaluated using various pollution indices which revealed that over 60% of the soils were between the high degree and the ultra-high degree of contamination classes. The concentration of various trace metals varies from 860.3–862.6 mg/kg for Cr; 324.9–328.4 mg/kg for Al; 200.9–203.4 mg/kg for As; 130.1–136.2 mg/kg for Fe; 121.9–125.8 mg/kg for Pb; 27.3–30.2 mg/kg for Co; 23.8–26.8 mg/kg for Ni; 7.2–9.2 mg/kg for Ti; 7.1–9.2 mg/kg for Cd; 4.0–5.6 mg/kg for Zn and 0.1–0.6 mg/kg for Cu

Diversity and antimicrobial activity of culturable fungal endophytes in Solanum mauritianum

CITATION: Pelo, S., Mavumengwana, V. & Green, E. 2020. Diversity and Antimicrobial Activity of Culturable Fungal Endophytes in Solanum mauritianum. International Journal of Environmental Research and Public Health, 17(2). doi:10.3390/ijerph17020439The original publication is available at https://www.mdpi.com/journal/ijerphPlant endophytes are microbial sources of bioactive secondary metabolites, which mimic the natural compounds chemistry of their respective host plants in a similar manner. This study explored the isolation and identification of fungal endophytes, and investigated the antibacterial and antimycobacterial activity of their crude extracts. Fungal endophytes were isolated from Solanum mauritianum, identified using morphological traits and internal transcribed spacer ribosomal-deoxyribonucleic acid (ITS-rDNA) sequence analysis. Eight fungal endophytes were identified as Aureobasidium pullulans, Paracamarosporium leucadendri, Cladosporium sp., Collectotrichum boninense, Fusarium sp., Hyalodendriella sp., and Talaromyces sp., while Penicillium chrysogenum was isolated from the leaves and unripe fruits. Good activity was observed for the crude extracts of Paracamarosporium leucadendri inhibiting Mycobacterium bovis, Klebsiella pneumoniae, and Pseudomonas aeruginosa at 6 µg/mL. Crude extracts of Fusarium sp., showed activity at 9 μg/mL against M. bovis, M. smegmatis and K. pneumonia. In general, the crude extracts showed great activity against Gram-negative and Gram-positive bacteria and novel results for two mycobacteria species M. bovis and M. smegmatis. The results provide evidence of diverse fungal endophytes isolated from Solanum mauritianum, and evidence that fungal endophytes are a good source of bioactive compounds with pharmaceutical potential, particularly against Mycobacterium tuberculosis.https://www.mdpi.com/1660-4601/17/2/439Publishers versio

Dataset on assessment of pollution level of selected trace metals in farming area within the proximity of a gold mine dump, Ekuhurleni, South Africa

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Antimycobacterial activity and molecular docking of methanolic extracts and compounds of marine fungi from Saldanha and False Bays, South Africa

The number and diversity of drugs in the tuberculosis (TB) drug development process has increased over the years, yet the attrition rate remains very high, signaling the need for continued research in drug discovery. In this study, crude secondary metabolites from marine fungi associated with ascidians collected from Saldanha and False Bays (South Africa) were investigated for antimycobacterial activity. Isolation of fungi was performed by sectioning thin inner-tissues of ascidians and spreading them over potato dextrose agar (PDA). Solid state fermentation of fungal isolates on PDA was then performed for 28 days to allow production of secondary metabolites. Afterwards, PDA cultures were dried and solid-liquid extraction using methanol was performed to extract fungal metabolites. Profiling of metabolites was performed using untargeted liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS). The broth microdilution method was used to determine antimycobacterial activity against Mycobacterium smegmatis mc2155 and Mycobacterium tuberculosis H37Rv, while in silico flexible docking was performed on selected target proteins from M. tuberculosis. A total of 16 ascidians were sampled and 46 fungi were isolated. Only 32 fungal isolates were sequenced, and their sequences submitted to GenBank to obtain accession numbers. Metabolite profiling of 6 selected fungal extracts resulted in the identification of 65 metabolites. The most interesting extract was that of Clonostachys rogersoniana MGK33 which inhibited Mycobacterium smegmatis mc2155 and Mycobacterium tuberculosis H37Rv growth with minimum inhibitory concentrations (MICs) of 0.125 and 0.2 mg/mL, respectively. These results were in accordance with those from in silico molecular docking studies which showed that bionectin F produced by C. rogersoniana MGK33 is a potential inhibitor of M. tuberculosis β-ketoacyl-acyl carrier protein reductase (MabA, PDB ID = 1UZN), with the docking score observed as −11.17 kcal/mol. These findings provided evidence to conclude that metabolites from marine-derived fungi are potential sources of bioactive metabolites with antimycobacterial activity. Even though in silico studies showed that bionectin F is a potent inhibitor of an essential enzyme, MabA, the results should be validated by performing purification of bionectin F from C. rogersoniana MGK33 and in vitro assays against MabA and whole cells (M. tuberculosis)

Toxic metal implications on agricultural soils, plants, animals, aquatic life and human health

Abstract:Theproblemofenvironmentalpollutionisaglobalconcernasitaffectstheentireecosystem. There is a cyclic revolution of pollutants from industrial waste or anthropogenic sources into the environment, farmlands, plants, livestock and subsequently humans through the food chain. Most of the toxic metal cases in Africa and other developing nations are a result of industrialization coupled with poor effluent disposal and management. Due to widespread mining activities in South Africa, pollution is a common site with devastating consequences on the health of animals and humans likewise. In recent years, talks on toxic metal pollution had taken center stage in most scientific symposiums as a serious health concern. Very high levels of toxic metals have been reported in most parts of South African soils, plants, animals and water bodies due to pollution. Toxic metals such as Zinc (Zn), Lead (Pb), Aluminium (Al), Cadmium (Cd), Nickel (Ni), Iron (Fe), Manganese (Mn) and Arsenic (As) are major mining effluents from tailings which contaminate both the surface and underground water, soil and food, thus affecting biological function, endocrine systems and growth. Environmental toxicity in livestock is traceable to pesticides, agrochemicals and toxic metals. In this review, concerted efforts were made to condense the information contained in literature regarding toxic metal pollution and its implications in soil,water,plants,animals,marine life and human health

Cytotoxic activity of crude extracts from Datura stramonium’s fungal endophytes against A549 lung carcinoma and UMG87 glioblastoma cell lines and LC-QTOF-MS/MS based metabolite profiling

Abstract : Background: Endophytic fungi are a proven source of bioactive secondary metabolites that may provide lead compounds for novel drug discovery. In this study, crude extracts from fungal endophytes isolated from Datura stramonium were evaluated for cytotoxic activity on two human cancer cell lines. Methods: Fungal endophytes were isolated from surface sterilized aerial parts of D. stramonium and identified using molecular, morphological and phylogenetic methods. Ethyl acetate crude extracts from these isolates were evaluated for cytotoxic activity on A549 lung carcinoma and UMG87 glioblastoma cell lines. Metabolite profiling was then performed by liquid chromatography coupled to quadrupole time-of-flight with tandem mass spectrometry (LC-QTOF-MS/MS) for the cytotoxic crude extract. Results: Eleven fungal endophytes were identified from D. stramonium. Significant cytotoxicity was only observed from the crude extract of Alternaria sp. KTDL7 on UMG87 glioblastoma cells (IC50 = 21.49 μg/ml). Metabolite profiling of this crude extract tentatively revealed the presence of the following secondary metabolites: 1,8-dihydroxynaphthalene (1), anserinone B (2), phelligridin B (3), metacytofilin (4), phomopsidin (5) and vermixocin A (6). Compounds 2 and 3 have been shown to be cytotoxic in literature. Conclusion: The findings in this study suggest that the crude extract of Alternaria sp. KTDL7 possesses compound(s) cytotoxic to glioblastoma multiforme cells. Future studies to isolate and characterize the cytotoxic compound(s) from this fungus could result in lead development of a fungal-based drug for glioblastoma multiforme treatment

The Relationship between Cadmium Toxicity and the Modulation of Epigenetic Traits in Plants

Elevated concentrations of heavy metals such as cadmium (Cd) have a negative impact on staple crop production due to their ability to elicit cytotoxic and genotoxic effects on plants. In order to understand the relationship between Cd stress and plants in an effort to improve Cd tolerance, studies have identified genetic mechanisms which could be important for conferring stress tolerance. In recent years epigenetic studies have garnered much attention and hold great potential in both improving the understanding of Cd stress in plants as well as revealing candidate mechanisms for future work. This review describes some of the main epigenetic mechanisms involved in Cd stress responses. We summarize recent literature and data pertaining to chromatin remodeling, DNA methylation, histone acetylation and miRNAs in order to understand the role these epigenetic traits play in cadmium tolerance. The review aims to provide the framework for future studies where these epigenetic traits may be used in plant breeding and molecular studies in order to improve Cd tolerance