DNA extraction of lipase-producing oryzae species from natural resources

Introduction and objective: Aspergillus refers to a genus of fungi which has been widely used in industry. Some of the species of this genus can contaminate agricultural crops such as pistachio and corn. In addition to destructive impacts on quality of the agricultural crops, they can result in poisoning and carcinogenity in living creatures including human. Rhizopus belongs to the family of opportunistic fungi and is categorized in zygomycetes branch; this fungus is also known as black bread mold. The vegetative mycelium of this fungus has a structure called sporangiophore. At the end of the sporangiophore, there is a dome-shaped body called columella. Vesicles are situated around the columella and are called sporangium. Inside the sporangium high numbers of sporia sporangio spores can be found. The aim of this study is to extract DNA of lipase-producing oryzae species from natural resources.Materials and methods: the samples were cultured on PDA. For enzyme production, mineral culture medium was used for solid state fermentation. The impact of carbon source, initial pH and incubation temperature on enzyme production was also addressed. After preparing spore suspension, solid state fermentation was carried out. The enzymes were extracted after fermentation and their enzymic activities were investigated. The samples’ DNA was extracted by boiling method, and their sequence was designed by specific primer. Sequencing was carried out after PCR.Results and discussions: most of Rhizopus oryzae genus species have high lipase activity and can produce lipase enzyme. Regarding abundance of this fungus in the country, it can be used as a significant genus in biotechnological and industrial activities. Based on present screening, the majority of local species can produce lipase. Rhizopus delemar strain BVKT R1and 8 Rhizopus oryzae isolate FPZSP368 have high lipase activity and can be regarded as promising isolates for production of this enzyme in large scales.

Keywords:

enzyme , DNA extraction , lipase , Aspergillus , fungus