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Assessment of waste frying oil transesterification capacities of local isolated Aspergilli species and mutants
Institution:1. Department of Chemistry of Natural and Microbial Products, National Research Centre, Dokki, Cairo, 12622, Egypt;2. Department of Genetics and Cytology, National Research Centre, Dokki, Cairo, 12622, Egypt;3. Department of Microbiology, Faculty of Science, Ain Shams University, Abbassia, Cairo, 11566, Egypt;1. Tochigi Prefectural Museum, 2-2 Mutsumi-cho, Utsunomiya, Tochigi, 320-0865, Japan;2. Hokken Co. Ltd., 7-3 Ekihigashimachi, Mibu, Shimotsuga-gun, Tochigi, 321-0222, Japan;3. Kanagawa Prefectural Museum of Natural History, 499 Iryuda, Odawara, Kanagawa, 250-0031, Japan;1. Graduate School, Khon Kaen University, Khon Kaen, 40002, Thailand;2. Biology Program, Faculty of Science, Buriram Rajabhat University, Buriram, 31000, Thailand;3. Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand;4. Department of Biochemistry and Biotechnology, Faculty of Agriculture, Tottori University, Tottori, 680-8553, Japan;5. Department of Microbiology, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand;1. Department of Marine Biodiversity Research, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka, Kanagawa, 237-0061, Japan;2. National Research Institute of Fisheries Science, Japan Fisheries Research and Education Agency, 2-12-4 Fukuura, Kanazawa, Yokohama, Kanagawa, 236-8648, Japan;3. National Museum of Nature and Science, Department of Geology and Paleontology, 4-1-1 Amakubo, Tsukuba, Ibaraki 305-0005, Japan;1. Faculty of Agriculture at Kamphaeng Sean, Kasetsart University, Kamphaeng Sean, Nakhon Pathom, 73140, Thailand;2. Department of National Park, Wildlife and Plant Conservation, Ministry of Natural Resources and Environment, Bangkok, 110900, Thailand;3. Department of Plant Pathology, National Chung Hsing University, Taichung, 402, Taiwan;4. Faculty of Education, Ibaraki University, Mito, Ibaraki, 310-8512, Japan;1. Graduate School of Sustainability Science, Tottori University, 4-101 Koyama-cho Minami, Tottori, 680-8553, Japan;2. Faculty of Agriculture, Tottori University, 4-101 Koyama-cho Minami, Tottori, 680-8553, Japan
Abstract:Biodiesel (fatty acids methyl esters, FAME) has attracted considerable attention as an environmentally and eco-friendly alternative for diesel engines. This research manipulates the use of Aspergillus whole-cell lipase as a biocatalyst for biodiesel production from waste frying oil (WFO). A total of 17 isolates of Aspergillus species screened for lipase and esterase production abilities. Qualitatively, 11 Aspergillus isolates showed lipase and/or esterase activities and only 4 isolates were able to perform WFO transesterification under the tested conditions. Two Aspergillus isolates showed relatively high FAME yields, thus were selected as good enzyme producers. These two isolates were molecularly identified using rRNA gene sequence ITS1 and ITS2 as A. tamarii NDA03a (Genbank Accession Number MK849615) and A. flavus NDA04a (Genbank Accession Number MK811208), respectively. These identified isolates were exposed to ethyl methanesulfonate (EMS) for producing hyper lipolysis mutants. Mutagenesis led to 13.15 and 14.45% improvement of the WFO transesterification by A. tamarii NDA03a and A. flavus NDA04a, respectively. Random amplified polymorphic DNA (RAPD) analysis of the produced mutants confirmed the genetic basis of the activity variation. Genetic polymorphism reached to 79.31% and 80.65% between A. flavus NDA04a and A. tamarii NDA03a mutants and their corresponding wild types, respectively.
Keywords:Biodiesel  EMS mutagenesis  RAPD analysis
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