Thesis (Ph.D. (Microbiology)) -- University of Limpopo, 2021 / Biofuel production from lignocellulose material is an attractive alternative to fossil fuel. The use of lignocellulose material for biofuel production is imperative because of the numerous advantages that it offers. Biofuel is environmentally friendly and in developing countries such as South Africa, it has the potential to reduce the use of imported fuel and create jobs. Currently, several constraints are affecting the implementation of biofuel. One of the constraints is the cost-effectiveness and the efficiency of the enzymes involved in the enzymatic degradation of lignocellulose polymers to monomers, which can further be fermented to bioethanol. The potential way to reduce enzymatic degradation cost could be by supplementing the fungal enzymes with accessory enzymes such as endo-xylanase. The enzyme production cost is also dependent on the carbon source used. Lignocellulose materials that are regarded as waste must be assed for their use as enzyme inducer carbon sources and as biomass for biofuel production. This is a potential route that will reduce enzyme and biofuel production costs. Biofuel production cost can further be reduced by finding a yeast that can ferment xylose and ferment in the presence of inhibitors released during lignocellulose pretreatment. This study sought to tackle the enzymatic hydrolysis constraints and also search for xylose-fermenting yeast by exploring the gut microbiota of dung beetle. The gut of the dung beetle has recently received great attention since it is proposed to be a bioreactor for lignocellulolytic microorganisms that can be used in biofuel applications. This is because dung beetles feed on the dung of herbivorous animals and the dung is composed of 80% undigested plant material. In this study the guts of four Scarabaeidae dung beetles Kheper nigroanaeus Boheman, Heteronitis castelnaui, Pachylomerus femoralis, Anachalcos convexus and dung beetle larvae, Euoniticellus intermedius were screened for hemicellulolytic fungi and xylose-fermenting yeast. Hundred and thirty-two yeast isolates and two-hundred and twenty-two filamentous fungi were isolated and identified using ITS and D1/D2 regions. The yeast isolates were assigned to 8 genera and 18 species, Trichosporon was the most dominant genus while Candida tropicalis was the most dominant specie. Some of the yeast isolates were identified as uncultured fungi. This yeast must be characterised to be certain if they are novel species. The fungal isolates were assigned to 12 genera and 25 species, Aspergillus was the most dominant genus while Hypocrea lixii was the most dominant specie. The yeast isolated could assimilate xylose and could grow at a maximum temperature of 40 °C. Furthermore, these yeast isolates could also grow in the presence of 3 g/L acetic acid. Most of the fungal isolates had xylanolytic activity. The phylogenetic analysis revealed close genetic relatedness between isolates from the different dung beetle species and dung beetle larvae. The profile of the fungal genera was similar in the different dung beetles. Both guts and the larvae had Aspergillus, Hypocrea, Trichoderma, Talaromyces and Penicillium. The filamentous fungi that showed good xylanolytic activity were further screened for their ability to produce xylanase enzyme using thatch grass as an inductive carbon source. Thatch grass was selected in this study since it is in-house plant-based biomass. Thatch grass is abundantly available in South Africa; it is used for animal grazing but the more it grows it loses its nutritional content. Once it reaches this stage, it is no longer used and most of it is burnt. The fire from burning grass contains higher levels of nitrogen-containing chemicals that pollute the environment. Its compositional analysis (cellulose 46%, hemicellulose 27% and lignin 10%) also attributed to its selection as potential inductive carbon and attractive lignocellulose biomass for biofuel production. The higher xylanase activity of 283.43, 270 and 287.03 nkat/ml were observed from Aspergillus fumigatus L1XYL9 (Euoniticellus intermedius larvae), Hypocrea lixii AB2A3 and Neosartotya sp AB2XYL20 (Anachalcos convexus), respectively. This was achieved when acid pretreated thatch grass was used as an inductive carbon source. Aspergillus fumigatus L1XYL9 (Euoniticellus intermedius larvae), Hypocrea lixii AB2A3 and Neosartotya sp AB2XYL20 (Anachalcos convexus) showed xylanase activity of 393,22, 313,06 and 200 nkat/ml when grown on synthetic xylan. Neosartotya sp AB2XYL20 showed higher xylanase activity on thatch grass. The suitable production process for xylanase enzyme on acid pretreated thatch grass was assessed by conducting a comparative study on solid-state and submerged fermentation using L1XYL9 (Euoniticellus intermedius larvae), Hypocrea lixii AB2A3 and Neosartotya sp AB2XYL20 (Anachalcos convexus) as the best xylanase producer on acid pretreated thatch grass. The strain showed better xylanase activity when submerged fermentation was used. In this study, Hypocrea lixii AB2A3 was selected for further studies since it was the most dominant species and also showed good xylanase activity. Thatch grass was pretreated differently to evaluate the suitable chemical for pretreating thatch grass. Thatch grass was pretreated with dilute sulphuric acid 1.2% and maintained the pH of 5.5 by using sodium hydroxide while another batch was pretreated the same way and was washed with distilled water till pH of 5.5. The other batch was then pretreated with ammonium solution and was also washed with distilled water to maintain a pH of 5.5. The above-mentioned pretreated thatch grass was tested as an inductive carbon source as well as untreated thatch grass. The xylanase activity was determined to assess a good inductive carbon. All the thatch grass pretreated and washed with distilled water showed very low xylanase activity. The untreated thatch grass resulted in lower xylanase activity as compared to xylanase activity achieved when pretreated thatch grass was used. Parameters such as agitation speed and initial inoculum size were also assessed during xylanase production by Hypocrea lixii AB2A3 on acid pretreated thatch grass. Xylanase activity increased from 525 nkat/ml (Inoculum size 2×106 spore/ml and agitation speed 150 rpm) to 584.8 nkat/ml (Inoculum size 2×106 spore/ml and agitation speed 200 rpm). The crude xylanase from Hypocrea lixii AB2A3 was used to hydrolyse acid pretreated thatch grass. This resultant in xylose yield of 138 mg/g of substrate and glucose yield of 49 mg/g of substrate. Crude xylanase was mixed with commercial celluclast™. This enzyme mixture resulted in a xylose yield of 128 mg/g substrate and a glucose yield of 549 mg/g of substrate. The results obtained in this study show that indeed gut of the dung beetles and dung beetle larvae are a rich source of microorganisms that can play an important role in biofuel application and remediating the environment by degrading plant-based biomass regarded as waste into valuable products. It is imperative to evaluate the gut microbiota of dung beetles from different regions in South Africa for their application in the biofuel industry to reinforce its implementation. Thatch grass is a potential inductive carbon and lignocellulose biomass for biofuel production. / NRF (National Research Foundation)

Thesis (MBA)--Stellenbosch University, 2006. / ENGLISH ABSTRACT: The adoption of the White Paper on the promotion of Renewable Energy and clean fuels in 2003, opened the playing field for participants from other industries than the conventional petroleum, to participate in the fuel industry in South Africa. South Africa is a net importer of crude oil, which accounts for 92% of liquid fuels supply in South Africa. Although the country has significant coal reserves which can supply the country's demand for approximately 200 years, this energy source contributes significantly to CO, emissions. South Africa's participation in the Kyoto Protocol compels it to abide by its commitments to reduce these emissions between 2008 and 2012. The country's dependence on energy to fuel its growing economy, and the infiationary impact that oil imports has had on the country's economy, has prompted government to explore alternative sources of energy to reduce its dependence on fossil fuels and especially importing crude oil. As a result of this, and in an attempt to increase the potential for the successful implementation of ASGISA, government is exploring the feasibility of introducing an E10 fuel blend to the South African petrol blend. In view of th is, government has in it Accelerated and Sustainable Growth Initiative (ASGISA) targeted the development of the bio-fuels industry as an industrial sector that presents opportunities to create opportunities for sustainable growth and development. In view of this, the fiedgling fuel-ethanol industry (which is in its construction phase at the t ime of writing this report), faces lucrative prospects for the agricultural industry, especially maize- and ethanol producers. It is anticipated that the fuel-ethanol industry will create between 8000 and 10000 direct and indirect employment opportunities per plant. This will result in significant investment in rural areas as well. The creation of employment in the rural areas will prevent the large-scale urbanisation that has become a phenomenon in the past decade, as a result of dwindling agricultural industries. The production of ethanol presents the opportunity to earn foreign exchange, especially if the industry embarks on large scale export strategies. In addition to the export market, the local market for ethanol consist of the possible E10 petrol-blend and to supply Eskom with ethanol to fuel its gas turbine electricity generators at Acacia, Port Rex, as well as the anticipated generators at Atlantis and Mossel Bay. This document is a report on the investigation of the economic evaluation of a bio-fuel industry in South Africa. It will explore the current outlook for fossil fuel reserves, supplies and demand, both internationally and locally. It will report on the phenomenon of peak oil production and some opinions thereon . An investigation into the most probable biomass that can be used as feedstock for bio-fuel production will conducted. In this regard, specific investigation into maize, sugar cane (for fuelethanol) and Jatropha eureas (for bio-diesel) will be conducted. The report will explore the most efficient ethanol production processes, for both maize- and sugar-to-ethanol production, with the weight of the document to be attributed to the economic impact that the adoption of the fuel-ethanol programme / AFRIKAANSE OPSOMMING: Die publisering van die Witskrif oor die promosie van hernieubare energiebronne en skoon brandstowwe in 2003, het die speelveld vir deelname aan die brandstof industrie oopgemaak vir rolspelers buiten die konvensionele petroleum maatskappye. Suid-Afrika is 'n netto invoerder van ru-olie en het in 2004 ongeveer 92% van die totale vloeibare brandstowwe ingevoer. Alhoewel die land aansienlike steenkool reserwes het om te voorsien in die aanvraag vir die volgende ongeveer 200 jaar, dra hierdie energiebron aansienlik by tot die koolstofdioksied vrystellings. Suid-Afrika se deelname aan die Kyoto Protokol van 1998, dwing die land om te voldoen aan die ondernemings wat gemaak is om hierdie koolstofdioksied vrystellings te verminder tussen 2008 en 2012. Die land se afhanklikheid van energiebronne om groei te stimuleer, asook die inflasionistiese effek van olie invoere op die ekonomie, het die regering genoop om alternatiewe bronne van energie te ondersoek sodat die afhanklikheid van olie verminder kan word. Uiteenlopend hiervan en om die implementering van ASGISA te stimuleer, ondersoek die regering tans die moontlikheid om 'n E10 petrol vermenging in die petrol formule te spesifiseer. Uit die oogpunt van ASGISA (Accelerated and Sustainable Growth Initiative) van Suid-Afrika, het die regering die ontwikkeling van die bio-brandstowwe industrie geoormerk om geleenthede te skep vir volhoubare ontwikkeling en groei. Met die oog hierop, voorspel die etanol bedryf, wat ten tyde van die skryf van hierdie verslag nog in kontruksie was, winsgewende potensiaal vir die landboubedryf, veral mielie produsente. Dit word verwag dat die etanol bedryf tussen ongeveer 8000 en 10000 direkte en indirekte werksgeleenthede sal skep, veral in die landelike gebiede. Dit sal grotendeels bydra tot die voorkoming van die voortslepende ontvolking van die platteland wat oor die afgelope jare 'n verlammende effek op plattelandse gebiede gehad het. Dit word ook voorsien dat daar aansienlike belegging in die platteland sal plaasvind en al hierdie faktore sal bydra tot die voorkoming van verstedeliking . Die etanol bedryf skep die geleentheid om buitelandse valuta te genereer, veral as die industrie op uitvoere gaan konsentreer. Indien 'n plaaslike mark beoog word , sal die implementering van die E10 vermenging 'n besliste mark skep. 'n Alternatiewe mark wat ondersoek kan word, en wat groot geleentheid skep, is Eskom, wat tans ingevoerde diesel verbruik om hul gas turbine krag opwekkers by Acacia en Port Rex van brandstof te voorsien . Indien die beoogde turbines by Atlantis en Mosselbaai gebou word, sal die mark vir plaaslike etanol verdubbel. Hierdie dokument is 'n verslag oor die ondersoek wat gedoen is na die lewensvatbaarheid van 'n brandstof etanol bedryf in Suid-Afrika. Dit berig oor die huidige uitkyk oor die fossiel brandstof reserwes in die wereld en plaaslik. Dit opper die vraagstuk oor piek olie produksie fenomeen wat uiteenlopende debate ontketen het. Die verslag dek die waarskynlike bronne van biomassa wat aangewend kan word in die produksie van etanol, met spesifieke verwysing na mielies, suikerriet en Jatropha curcas. Die mees effektiewe produksie metodes word verder ondersoek wat van toepassing is op beide mielies en suikerriet. Die mees relevante deeI van die verslag is die ondersoek na die ekonomiese impak wat die industrie op die Suid-Afrikaanse ekonomie mag hê, waarna die nodige gevolgtrekkings en aanbevelings gemaak sal word.

Historically, the most frequently used energy sources have been those nearest and easiest to consume. Unfortunately, society’s reliance on fossil fuel for power generation has occurred at the expense of the environment, coal being a major contribution to carbon dioxide (CO2) emission. Carbon dioxide is classified as a greenhouse gas (GHG); it contributes to the phenomenon of climate change (Haw & Hughes, 2007, p.1). According to Worrell (2011), industry uses nearly 40 percent of worldwide energy on economic activities. Value chain activities alone contribute almost 37 percent to global GHG. Organisations are socially and ethically required to minimise the carbon footprint of their operations. Reducing energy use makes perfect business sense; it saves money, enhances corporate reputations and helps everyone participate the fight against climate change (Carbon Trust, 2011). Gielen, Newman, and Patel (2008) strongly believe the overall energy and emissions trends can be mitigated through additional energy efficiency measures. However, implementing EnMS will enable organisations to establish systematic approaches and the processes necessary to improve energy performance, including energy efficiency, use and consumption (SANS 50001, 2011). The objective of this paper was to evaluate the current energy management strategy adopted by selected automotive manufacture in Eastern Cape. The research was motivated by the fact that previous researchers have focused more on technological aspects and less of management functions. The research paradigm followed in this paper was qualitative because a case study is used to gain an insight and understanding about more and less successful energy management strategies. In this report, background about the global energy outlook and its significant to economic development, factors behind energy demands, the link to climate change and providing effective energy management principles are covered. The energy management principles covered key elements for delivering successful energy management. Literature highlighted that, senior management commitment is the foundation of good energy management, which is delivered through a formal energy policy and a supporting energy strategy with action plan. High level commitment will provide: Advocacy from senior managers; Visibility of the issues across your organization; Impetus for the organisation to implement energy management; Resources, both human and financial. It will also demonstrate that good energy management is part of your organisation’s mission and as relevant as other management aspects. The empirical study is focused on the characteristics of the current management system and organisational structure employed with its relevant functions. Based on these reference points the paper concludes with recommendations for the case study organisation.