The influence of oxidized oils on fungal growth and lipid utilization

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Date
2010-03
Authors
Leeuw, Ntsoaki Joyce
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University of the Free State
Abstract
English: Edible oils such as sunflower oil, soybean oil and palm oil are used today in the frying of food. During the frying process, various changes such as removal of antioxidants, hydrolysis, oxidation and polymerization occur in these oils. These reactions are responsible for a variety of physical and chemical changes observed in the oil during frying and may lead to the formation of breakdown products which include polar compounds (PCs) and polymerized triglycerides (PTGs). South African regulations state that oils that contain 16% and more PTGs and 25% and more PCs are harmful to human health. These oils may cause cancer and diarrhoea in humans and animals. However, little is known regarding the effect of oxidized oils on fungi. The oleaginous fungi Cryptococcus curvatus and Mucor circinelloides were used to determine the effect of palm oil breakdown products, measured as PTGs on lipid turnover, growth and morphology. In Mucor circinelloides we found, after seven days of growth, a decrease in biomass, lipid utilization and accumulation at increased PTG levels, at low and neutral pH. An increase in PTG concentration also influenced the morphology of M. circinelloides. Protrusions were observed on cell surfaces when grown on oil with 45% PTGs and not when the fungus was grown on fresh oil with 0.4% PTGs. In C. curvatus there was also a decrease in oil utilization and biomass production at increased PTG levels, at low and neutral pH. An increase in oil accumulation was observed at low pH while it remained constant at neutral pH for all PTG levels tested. Hairy and warty protuberances on cell surfaces were observed when C. curvatus was grown on oils with 15% and 45% PTGs, respectively. It is concluded that the changes observed in lipid turnover and morphology in both fungi are due to the presence of palm oil breakdown products. Oxidized oil breakdown products such as aldehydes are major sources of reactive oxygen species (ROS). Studies have shown that ROS has anti-mitochondrial action. It was also reported that acetylsalicylic acid (ASA), an anti-inflammatory and anti-mitochondrial drug, targets structure development and functions of yeasts, needing elevated levels of mitochondrial activity. Using antibody probes it was previously reported that sporangia of Mucor circinelloides also contain increased mitochondrial activity yielding high levels of 3-hydroxy (OH) oxylipins. This was however not found in Mortierella alpina (subgenus Mortierella). In this study, it is reported that oxidized palm oil breakdown products and ASA also targets sporangium development of M. circinelloides selectively while hyphae, with lower levels of mitochondrial activity, are more resistant. Similar results were obtained when the anti-inflammatory compounds benzoic acid, ibuprofen, indomethacin and salicylic acid were tested. Here, oxidized oils and antiinflammatory, anti-mitochondrial drugs exerted similar effects on this dimorphic fungus as found under oxygen limited conditions. Interestingly, sporangium development of M. alpina was found not to be selectively targeted by these compounds. Mortierella alpina, which could not expose dimorphic growth under oxygen limitation conditions, was also more sensitive to the anti-inflammatory drugs when compared to M. circinelloides. These results prompt further research to assess the applicability of these anti-mitochondrial antifungals to protect plants and animals against Mucor infections. It is concluded that indications exist that oxidized palm oil breakdown products target mitochondrial function. This may explain the inhibitory effect of these compounds on fungal growth, lipid turnover and altered cell wall morphology.
Afrikaans: Eetbare olies, soos sonneblom-, soja- en palmolie is van die olies wat vandag in die braai van voedsel gebruik word. Gedurende die braaiproses vind verskeie veranderinge, soos die verwydering van anti-oksidante, hidrolise, oksidasie en polimerisering, in hierdie olies plaas. Hierdie reaksies is verantwoordelik vir ‘n verskeidenheid fisiese en chemiese veranderings wat in die olie tydens die braaiproses waargeneem word en wat mag lei tot die vorming van afbraakprodukte, insluitend polêre komponente (PKs) en gepolimeriseerde trigliseriede (PTGs). Suid-Afrikaanse wetgewing bepaal dat olies wat 16% en meer PTGs en 25% en meer PKs bevat, skadelik vir menslike gesondheid is. Hierdie olies mag kanker en diariëe in mense en diere veroorsaak. Daar is egter nog min bekend oor die effek van geoksideerde olies op fungi. In hierdie studie, is die oleogene fungi Cryptococcus curvatus en Mucor circinelloides gebruik om die effek van palmolie-afbraakprodukte, gemeet as PTGs, op lipiedomset, groei en morfologie te bepaal. Na sewe dae van groei, het ons in Mucor circinelloides ‘n afname in biomassa, lipiedverbruik en –ophoping by verhoogde PTG-vlakke gevind, by lae en neutrale pH. ‘n Toename in PTG-konsentrasie het ook die morfologie van M. circinelloides beïnvloed. Uitsteeksels is op die seloppervlakte waargeneem na groei op olie met 45% PTGs en nie nadat die fungus op vars olie met 0.4% PTGs gegroei is nie. Daar was ook ‘n afname in olieverbruik en biomassa produksie by C. curvatus by verhoogde PTG-vlakke, by lae en neutrale pH. ‘n Toename in olie-ophoping is by lae pH waargeneem, maar by neutrale pH het dit konstant gebly vir alle getoetsde PTG-vlakke. Harige en vratagtige uitsteeksels is op die seloppervlaktes waargeneem nadat C. curvatus op olies met onderskeidelik 15% en 45% PTGs gegroei is. Ons lei af dat die veranderinge in lipiedomset en morfologie wat by beide fungi waargeneem is, die gevolg is van die teenwoordigheid van palmolie-afbraakprodukte. Afbraakprodukte soos aldehiede is hoofbronne van reaktiewe suurstofspesies (RSS). Studies het aangetoon dat RSS antimitochondriale werking het. Dit is bekend dat asetielsalisielsuur (ASS), ‘n antiïnflammatoriese en antimitochondriale middel, die ontwikkeling van strukture en funksies in giste, wat afhanklik is van verhoogde vlakke van mitochondriale aktiwiteit, teiken. Dit is voorheen getoon dat sporangia van Mucor circinelloides ook verhoogde mitochondriale aktiwiteit het, wat lei tot hoë vlakke van 3-hidroksie oksilipiene toon. In hierdie studie is bevind dat ASS ook sporangiumontwikkeling in M. circinelloides selektief teiken, terwyl hifes, met laer mitochondriale aktiwiteit, meer weerstandbiedend is. Soortgelyke resultate is verkry toe die antiïnflammatoriese verbindings, benzoësuur, ibuprofen, indometasien en salisielsuur getoets is. In hierdie studie is geoksideerde olies (met hoë PTG-vlakke) ook ingesluit. Hierdie olies is beweerde inhibitore van mitochondriale aktiwiteit. Hier het antiïnflammatoriese middels en geoksideerde olies soortgelyke effekte op hierdie dimorfe fungus gehad as toestande van suurstofbeperking. Merkwaardig is bevind dat sporangiumontwikkeling in Mortierella alpina nie selektief deur hierdie verbindings geteiken word nie. Mortierella alpina, wat nie dimorfies onder suurstofbeperking groei nie, was ook meer sensitief vir antiïnflammatoriese middels in vergelyking met M. circinelloides. Hierdie resultate verg verder navorsing om die toepaslikheid van hierdie antimitochondriale antifungale middels om plante en diere teen Mucor infeksies te beskerm, te bepaal. Hierdie studie impliseer dat geosideerde palmolie-afbraak produkte, mitochondriale funksie teiken. Dit kan moontlik die inhibitoriese effek van hierdie komponente op fungale groei, lipiedomset en veranderde selwandmorfologie verklaar.
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Keywords
Palm oil, Mitochondria, Lipids, Sporangia, Polymerized triglycerides, Oxidized oils, Mucor circinelloides, Morphology, Anti-mitochondrials, Cryptococcus curvatus, Lipid turnover, Mitochondrial activity, Anti-inflammatory compounds, Thesis (Ph.D. (Microbial, Biochemical and Food Biotechnology))--University of the Free State, 2010
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