M. Tech. (Chemistry, Faculty of Applied and Computer Sciences), Vaal University of Technology. / Advanced oxidation processes (AOPs) are supposedly effective means for removal of low concentration of organic pollutants from waste water as compared to conventional treatment methods. However, TiO2 metal semiconductor is the most promising photocatalyst for degradation of organic pollutant under heterogeneous photocatalysis as compared to other metal semiconductors. Challenges such as aggregation in solution, low adsorptive ability for non-polar organic contaminants and recycling are limitations in application of TiO2 for commercial purposes. The other limitations of TiO2, is it only utilizes 4-6% of the solar energy reaching the earth's surface which is in the UV region and also rapid electron-hole recombination due its wide band gap. In this work, the limitations are overcome by synthesis of a new photocatalyst material and further applied on degradation of model organic contaminants. The first part of this work focused on preparation and characterization of photocatalyst material. The photocatalyst synthesized were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray analysis (EDAX), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA) and UV-VIS diffuse reflectance spectrophotometer (DRUV-VIS). Supporting characterization techniques revealed partly dispersion of TiO2 within the cavities of dealuminated Clinoptilolite (HCP). TiO2 exist as nanoparticles or clusters on the HCP surface ascribed to lower loading of TiO2. XRD analysis showed that the support material employed was mainly Clinoptilolite and absorption band of prepared photocatalyst was red-shifted into the visible region, with slight reduction in band gap of photocatalyst. The second part focused on adsorption and photocatalytic degradation of methyl orange solution (MO) conducted under UV-irradiation in the presence of TiO2/HCP. The influence of operational parameters on degradation efficiency of photocatalyst material on MO was carried out in this study. Parameters such as initial dye concentration, pH, calcination temperature, inorganic anions and peroxide concentration were varied during degradation activities of MO. Comparative degradation efficiency of TiO2/HCP, TiO2 and HCP were conducted on dye mixture (Methyl orange and Methylene Blue) under UV irradiation. Kinetic analysis employing Langmuir-Hinshelwood model on dependencies of organic contaminants degradation was also conducted at different operational parameters. The adsorption capacity of MO was highest in the presence of TiO2/HCP at lower loading, which is ascribed to good dispersion of TiO2 on HCP and increased surface area of dealuminated Clinoptilolite. The photocatalytic degradation of methyl orange in the presence of TiO2/HCP was optimized at low dye concentration (30 ppm), acidic condition (pH 4), and calcination temperature of 873 K. Nitrate ion of Sodium salt accelerates degradation activities on methyl orange as compared to other inorganic anions. Photocatalytic degradation of methyl orange was greatly enhanced upon addition of oxidant (H2O2) and the photocatalyst possessed good repeatability after 3 cycles. TiO2/HCP exhibit highest degradation activities, followed by HCP as compared to TiO2 during the degradation of dye mixture. The degradation of MO by the photocatalyst fits into pseudo-first order kinetic model, while for comparative analysis of photocatalyst on dye mixtures follows second order kinetic model.

Thesis (MTech (Food Technology))--Cape Peninsula University of Technology, 2018. / Lipase-catalysed chemical transformations are today routinely considered by synthetic organic chemists as economical and competitive “green chemistry” alternatives. Although lipases can effortlessly be produced on a large-scale by fermentation, their industrial application was, until recently, limited to the detergent, oleo-chemistry and dairy industry. However, during the last few decades, the biotechnological application of lipases has expanded significantly, becoming indispensable in the manufacture of pharmaceuticals, pesticides, single cell protein production, biosensor preparations and waste management. Similarly, lipases have become a vital ingredient in the contemporary food processing industry with applications ranging from fruit juice production to baked foods, vegetable fermentations and dairy enrichment. Furthermore, lipases are routinely used as flavour development agents in cheese, butter and margarine products. Lipases are also applied in the leather industry for processing hides and skins and for treatment of activated sludge and other aerobic waste product treatments where its action enhances oxygen transfer. While lipases currently account for less than 21 % of the enzyme market, a growing interest in lipases is reflected by the publication of an average of 1000 research papers per year and the growing number of available lipases since the 1980s. There is a sustained interest to bioprospect for novel lipase enzymes from available unexplored biodiversity. This study aimed to screen for lipase-producing microorganisms resident in olive wastewater biofilms. Lipase activity of positive isolates was subsequently also quantitatively determined to select for the highest producers of true lipases. A Geotrichum candidum isolate from olive mill wastewater biofilms was selected for subsequent studies based on its superior lipase production phenotype. Using a yeast mediated ligation approach the G. candidum GCL1 lipase gene was cloned and heterologously expressed in Saccharomyces cerevisiae as an enzyme production host. The recombinant lipase was purified and analysed in terms of substrate specificity, pH optima, temperature optima and stability as well as organic solvent tolerance. The G. candidum gcl1 lipase presented enhanced thermo- and organic solvent-stability that are highly sought after traits for industrial application.

Thesis (D.Tech.: Biotechnology)-Dept. of Biotechnology, Durban University of Technology, 2008. xxiv, 322 leaves / Activated sludge models now in use worldwide for the design and operation of treatment systems use hypothetical concentrations of active organisms. In order to validate and calibrate model outputs, concentrations and activities of organisms responsible for nitrification and denitrification need to be reflected by actual measurements. This research has been initiated by the observation of an increasing gap of suitable techniques that exist in the direct measurement and separation of active biomass components, responsible for COD removal and denitrification.

Interacting microbial associations capable of utilizing thiosulphate as an energy source were enriched/isolated from activated sludge, landfill site [mal covering soil and soil from an acid mine water drainage site. The isolates were designated Lf-I, Ws-2 and Am-3, respectively. Although hydrogen sulphide was the target molecule for gas biofiltration, thiosulphate, which is a key oxidized intermediate, was used in this study due to the difficulty of working with a toxic gas. Together with thiosulphate oxidation, the microbial associations were assessed for their abilities to oxidize dissolved sulphide to elemental sulphur. Physiological analyses (temperature, pH and substrate concentration optimization) were made with closed and open cultures while morphological characterization and species compositional changes were monitored by light and scanning electron microscopy (SEM). To investigate further functional and structural responses to physiological changes, denaturing-gradient gel electrophoresis (DGGE) separation of PCR-amplified 16S DNA gene fragments and Biolog GN microtitre plates were used. The associations were found to be active metabolically between 0 and 35°C, 15 and 50°C, and 15 and 45°C, with optimum temperatures of 25, 40 and 35°C for Lf-l, Ws-2 and Am-3, respectively. The optimum pH range for microbial association Lf-l was between 3 and 4. The maximum specific growth rates of associations Lf-l , Ws-2 and Am-3 were 0.08, 0.06 and 0.03 h~l , respectively. Components of all three Gram negative rod-dominated associations were motile and displayed anaerobiosis. During open culture cultivation the species complement of Lf-l , as determined by morphological analysis, changed. The same association oxidized sulphide (40 ppm) to sulphur although Ws-2 and Am-3 did not have this capacity. Biolog GN plates detected pH-effected species compositional changes in Lf-l and these were confirmed by DGGE. The same technique showed that enrichment had occurred in the Biolog GN wells. Species composition changes also resulted in response to different pH values (2 to 9), temperatures (5 to 40°C) and dilution rates (0.003 to 0.09 h-1 ), but activity changes were not always accompanied by population profile changes. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2000.

Fermentation is a process by which primary food products are modified biochemically by the action of microorganisms and/or their enzymes. Several societies have, over the years, intentionally carried it out to enhance the taste, aroma, shelf-life, texture, nutritional value and other properties of food. It is used in many parts (lithe world. However, there are regional differences in use and these depend on the availability of raw materials, consumption habits. and other socio-cultural factors. This study was aimed at (comparatively) assessing, local commercial and homemade amahewu with respect to nutritional value, hygiene and other health benefits to the commirn ity. Methods employed were Thin Layer Chromatography (TLC) (mycotoxins), High Perliffmance Liquid Chromatography (HPLC) (mycotoxins, sugars and amino acids), Dumas (proteins), SOxhlet (lipids) and intubation technique (metabolisable energy) to analyse maize meal and amahewu samples from various regions. The regions sampled included mal3heleni (South Coast) and kwaNgcolosi (North Coast) villages. Commercial amahewu was analysed with kind permission from Clover SA. Species from the following genera were isolated and identified from amahewu samples: Lactobacillus, Saccharonivccs, Lcuconostoc, Lactococcus, Panioca, Entcrobacter and kleb•iella. Saccharotnyces was detected in commercial samples only. Gram-negative strains were identified in most of manheleni village samples. No traceable amounts of aflatoxin BI (AFB1), fumonisin B 1 (FBI) and zearalenone (ZEA) were found in Clover SA samples. AFB I was detected in 40% of both maize meal and amahewu samples from maBheleni (range 0.55 — 0.84ng/g and 8.3x10 5 — 9.1x10-5ng/g respectively). From the same village, 100% of the maize meal and 80% of the amahewu samples were contaminated with FBI (range 4.1 47.2ng/g and 1.4 ---- 6.9ng/g respectively). ZEA was detected in all maize meal samples (range 0.9 — 4.3ng/g). None of the amahewu samples contained detectable levels of ZEA. All maize meal and amahewu samples from kwaNgcolosi were contaminated with AF13 1 (range 8.3 — 30.I ng/g and 0.04 - 0.102ng/g respectively). FB I was detected in 75% of both maize meal and amahewu samples from the same village (range 0.5 — 4.1ng/g and 0.04 0.56ng/g respectively). ZEA was also found in all maize meal samples and 75% of amahewu samples (range 3.7 — 16.4ng/g and 0.03 -- 0.06ng/g respectively). MaBheleni, Clover SA and kwaNgcolosi maize meal and amahewu samples contained vitamins B1, 13 2 and B6 with a range of 0.31+0.21 - 4.48±0.81 B 1 ; 0.15±0.14 - 1.67±0.33 B2 and 0.05±0.07 - 0.77±1.45 lig/g B6. Fat levels ranged from 0.28±0.40 to 4.54±0.05 percentage by weight. The levels of proteins varied from 4.02±0.02 to 8.40±0.04 percentage by weight. Starch concentrations ranged from 31.51.5.28 to 75.911.92g/100g. Maize meal samples contained glucose and maltose, while glucose, fructose, sucrose, maltose, M-triose, DP 4 and 5 and DP >15 were detected in amahewu. Apparent and true metabolisable energy for homemade and commercial Freeze-dried amahewu was 13.194 and 13.696MJ/kg (AME N ); and 13.605 and 14.106M.Ekv ( 1 MEN ), respectively. This study has shown that lactic acid maize fermentation reduce' the levels of AF13 1 , FB I and ZEA toxins in maize meal, inhibits the growth of most Gram-negative bacteria, and in some instances, fermentation did improve the nutritional value. Metabolisable energy analysis represents an important tool to assess whether or not compounds ingested are converted to sources of energy in the body and utilised. Amahewu fermentation yielded beneficial products (probiotics: reduced mycotoxins levels and reduced starch). In conclusion, natural lactic acid maize fermentation to produce amahewu will do more good than harm to the consumer, therefore, people need to be advised on how to safely store their maize and also to be encouraged to consume their stored maize in fermented form. / Thesis (M.Med.Sc.)-University of Natal, Durban, 2003.

Microbial fuel cell (MFC) systems capable of both treating wastewaters and recovering energy have the potential for successful scale-up as a low carbon technology. These systems utilize microorganisms residing in biofilms as biocatalytic agents in the conversion of reduced substrates to electrical energy. As such, it is important to understand how MFC anodic biofilms develop over time and also how environmental parameters such as substrate type, temperature, carbon support material, anode architecture and optimized applied potentials also affect electrogenic performance. The type of substrate was found to have a large impact on the acclimation and performance of electrogenic biofilms. Acetate produced the highest power density of 7.2 W m 3 and butyrate the lowest at 0.29 W m"3, but it was also found that biofilm acclimation to these different trophic conditions also determined the MFC response to different substrate types i.e. both acetate and butyrate substrates produced power densities of 1.07 and 1.0 W m"3 respectively in a sucrose enriched reactor. The use of MFCs for wastewater treatment in temperate regions requires the development of reactor systems that are robust to seasonal fluctuations and are energy efficient. As such, system performance was examined at three different operating temperatures (10°C, 20°C and 35°C). At each temperature a maximum steady-state voltage of 0.49 V ± 0.02V was achieved after an operational period of 47 weeks, with the time to reach steady-state voltage being dependent on acclimation temperature. The highest COD removal rates of 2.98g COD L^d * were produced in the 35°C reactor but coulombic efficiencies (CE) were found to be significantly higher at pyschrophilic temperatures. Acclimation at different operating temperatures was found to a have a significant effect on the dynamic selection of psychrophilic, psychrotolerant and mesophilic anode respiring bacteria (ARB) and also influence the development of biofilm biomass, methanogenesis and electrogenic activity. Although start-up times were inversely influenced by temperature the amount of biomass accumulation increased with higher operational temperatures and this had a direct impact on biocatalytic performance. The three dimensional structure and porosity of different carbon anode materials affected anodic performance by determining the levels of surface area available for biofilm growth and the capacity for mass transfer to occur. Novel helical electrode configurations were used to look at the effect of altering turbulent flows to increase mass transfer rates and carbon surface areas available for electrogenic growth. The spiral with the highest amount of carbon veil and the smallest gap produced the highest power production of 11.63 W m"3 . Comparative studies of a logic controlled and un-controlled external load impedance showed that control affected the biocatalyst development and hence MFC performance. The controlled MFC better optimized the electrogenic anodic biofilm for power production, indicating that improved power and substrate conversion can be achieved by ensuring sustainable current demand, applied microbial selection pressures and near-optimal impedance for power transference.

Orientador: Denise Bevilaqua / Banca: Ana Teresa Lombardi / Banca: Paulo Teixeira Lacava / Resumo: A demanda crescente de metais como ferro, cobre, entre outros ocasionou o esgotamento progressivo dos depósitos minerais ; de tal modo que as companhias minera doras desenvolver am tecnologias alternativas aos métodos convencionalmente aplicados para a recuperação e extração de metais valiosos partir de minerais de baixo teor e outras fontes de metais polimetálicos como minerais de ferro que contem metais de base associados . Uma dessa s alternati vas é a biomineração que utiliza principalmente microrganismos procariontes e eucariontes para acelerar a dissolução oxidativa de minerais sulfetados presentes em minér i os de baixo teor produzindo a solubilização de metais associados; esta biotecnologia é atualmente usada apenas para processar minérios reduzidos e rejeitos minerais uma vez que a maioria destes minerais são sulfetados. No entanto, muitos metais de valor econ ômico também são encontrados em minerais que são parcial ou totalmente oxidados como as lat eritas de cobre ou níquel, minerais que não são susceptíveis à dissolução oxidativa; portanto, os minerais de ferro férrico contidos nes ses minérios não podem ser pr ocessados oxidativamente, pois estes compostos são susceptíveis de serem reduzidos por processos biológicos, derivando na solubilização de metais associados . Microrganismos acidófilos tais como Acidithiobacillus ferrooxidans podem catalisar a redução dissimilatória de íons férrico s na ausência de oxigênio para acelerar a solubilização destes metais. Estud os recentes têm mostrado que ess a nova abordagem pode ser utilizada para extrair metais tais como níquel e cobre a partir d e minérios oxidados a uma velocidade mais elevada do que pode ser conseguido por processos oxidativos. Neste trabalho, foram realizados ensaios de redução biológica de íons férrico s acoplado a oxidação anaeróbia de enxofre elementar em um biorreator... / Abstract: The growing demand for metals such as iron, copper, and others has caused the gradual exhaustion of mineral deposits; such a way that the mining companies have developed technologies alternative to the methods conventionally applied for the recovery and e xtraction of valuable metals from low grade minerals and other sources of polymetallic metals such as iron minerals which contains base metals associated . One such alternative is the biomining which uses mainly microorganisms prokaryotes and eukaryotes to accelerate the oxidative dissolution of sulphide minerals present in low grade ores producing solubilization of associated metals; This biotechnology is currently used only to process reduced ores and minerals tailings since most of these minerals are sulp hides. However, many metals of economic value are also found in minerals that are partially or fully oxidized like copper or nickel laterites, minerals that are not susceptible to oxidative dissolution; therefore, the mineral ferric iron contained in these ores can not be oxidatively processed, since these compounds are capable of being reduced by biological processes, deriving the associated solubilizing metals. Acidophilic microorganisms such as Acidithiobacillus ferrooxidans can catalyze the dissimilator y reduction of ferric iron in the absence of oxygen to accelerate the solubilization of these metals. Recent studies have shown that this new approach can be used to extract metals such as copper and nickel from oxide ores at a higher speed than can be ach ieved by oxidative processes. In this work were carried out biological reduction tests of ferric iron coupled anaerobic oxidation of elemental sulfur in an automated bioreactor 2 L with temperature, agitation and pH constant in anaerobic conditions using a n iron ore and a pure culture of facul tative anaerobic bacterium At. ferrooxidans. The sample was provided by the Vale Technology... / Mestre

Please read the abstract in the front section of this document / Dissertation (MSc (Microbiology))--University of Pretoria, 2005. / Microbiology and Plant Pathology / unrestricted

Current energy and environmental challenges are driving the use of cellulosic materials for biofuel production. A major obstacle in this pursuit is poor ethanol tolerance among cellulolytic Clostridium species. The first objective of this work was to establish a potential upper boundary of ethanol tolerance for the cellulosome itself. The hydrolytic function of crude cellulosome extracts from C. cellulolyticum on carboxymethyl cellulose (CMC) with 0, 5, 10, 15, 20 and 25% (v/v) ethanol was determined. Results indicated that the endoglucanase activity of the cellulosome incubated in 5% and 10% ethanol was significantly different from a control without ethanol addition. Furthermore a significant difference was observed in endoglucanase activity for cellulosome incubated in 5%, 10%, 15%, 20% and 25% ethanol in a standalone experiment. Endoglucanase activity continued to be observed for up to 25% ethanol, indicating that cellulosome function in ethanol will not be an impediment to future efforts towards engineering increasing production titers to levels at least as high as the current physiological limits of the most tolerant ethanologenic microbes. The second objective of this work was to study bioethanol production by a microbial co-culture involving Clostridium cellulolyticum and a recombinant Zymomonas mobilis engineered for the utilization of oligodextrans. The recombinant Z. mobilis ZM4 pAA1 and wild type ZM4 were first tested on RM medium (ATCC 1341) containing 2% cellobiose as the carbon source. Ethanol production from the recombinant Z. mobilis was three times that observed from the wild type Z. mobilis. Concomitant with ethanol production was the reduction in OD from 2.00 to 1.580, indicating the consumption of cellobiose. No such change in OD was observed from the wild type. The recombinant ZM4 was then co-cultured with C. cellulolyticum using cellobiose and microcrystalline cellulose respectively as carbon sources. Results indicate that the recombinant ZM4 acted synergistically with C. cellulolyticum to utilize 2.0 g L-1 cellobiose, producing as much as 0.40 mM concentration of ethanol whereas only 0.20 mM ethanol was detected for the wild type ZM4 co-cultured with C. cellulolyticum under the same conditions. A co-culture of the recombinant ZM4 and C. cellulolyticum using 7.5 g L-1 microcrystalline cellulose gave lower ethanol yield than when using cellobiose. In the latter case, the recombinant began producing ethanol in 5 days whereas the wild type required 10 days to produce detectable ethanol. Future efforts will concentrate on identifying the correct concentration of cellulosic substrate at which synergy will be observed using the recombinant ZM4 and other cellulose degrading microorganisms, as well as optimizing medium formulations to better support both organisms.

This master thesis deals with the development of a minibioreactor for microbial biotechnologies. The AlgaTox system – an analytical photobioreactor from BVT Technologies – was the default unit of the new device. The working volume of the new minibioreactor is in the range of 4 to 8 ml. The minibioreactor was composed of a minithermostat, a reaction vessel, an oxygen electrode, a temperature and pH probe, accessories for the supply of air to the liquid and an insertion for the transport of liquid from / to the reactor. The functionality of the assembled device and its characteristics were measured. Furthermore, an operating procedure for the decontamination process using a hydrogen peroxide mist was developed for the new equipment. An operating procedure for culturing microorganisms in a minibioreactor was also set up. Pilot cultivations of Halomonas halophila were demonstrated in the prepared equipment. In these cultivations, three assemblies for air supply to the liquid were tested. However, none of the assemblies was able to provide a sufficient supply of oxygen to cell culture – the dissolved oxygen value always dropped to 0%. For the further development, a new assembly was designed for the supply of air to the liquid - an aeration ring from a membrane tube.