Isolation, Identification And Lipase Gene Cloning Of Highly Efficient Petroleum Degrading Strains Of Marine Filamentous Fungi

The oceans are important to the survival of humans resource, Marine oil exploitation and transportation is often Marine pollution caused by oil spill.According to preliminary statistics, nearly four decades along the coast of China(1973-2011), some 3000 of ship oil spill accident, the average 4-5 days a pollution accident occurs.Among them, the disposable leak 95, 50 tons of oil spill by an average of 2.5, the average pollution accident-- 537 tons, total of 38500 tons of oil spill.In penglai 19-3 oil spill accident, for example, a large number of petroleum pollutants into the water and sediment, caused the penglai 19-3 oil field and its surrounding a total of 870 square kilometers northwest waters severely contaminated seawater, the water content of full 53 times more than the background value.The oil spill accident results in the decrease of the phytoplankton species and diversity of Marine pollution, Marine biological larvae damage of larvae and eggs larvae, benthic organisms petroleum hydrocarbon content increased significantly, Marine biological habitat destruction, the 2014 China ocean environment bulletin shows that it was not until 2014 that the impact of oil spill is still there.Therefore, research on Marine oil pollution science quickly become the focus of scholars both at home and abroad.Bioremediation is environmental protection, no pollution, safe and reliable again of Marine oil pollution treatment measures, to obtain the efficient Marine oil pollution degradation strains is an important link.Due to sea water belongs to the high salt environment, offshore oil degradation bacteria must have resistance to salt, easy to float the water and oil combined with characteristics, and oil as carbon source and use its ability to survive.Practice has proved that may have the above three characteristics of filamentous fungi, therefore screening in the sea is the precondition of leaking oil processing of filamentous fungi.This study We got 4 filamentous fungi degradating petroleum from ocean waters the oil pollution polluted by petroleum in the ocean waters to get 4 of oil degradation filamentous fungi, with one of the fast growth of fungus strains as the material, the esterase gene cloning and prokaryotic expression, won the highly active reorganization of esterase.The main results were as follows:1.the bohai bay oil pollution in Marine fungi were separated.To use oil as sole carbon source of culture medium, 4 strains screened in the available oil fungus of survival.By observation of biology, IST sequence amplification and sequencing identification, identify the 4 kinds of fungi to red powder capsule(Eurotium rubrum SH58), broadsword sickle bacteria(Fusarium culmorum S21), glue spore anthrax(Colletotrichum gloeosporioides SH4) and tiny hair mildew(Rhizomucor pusillus CC2).Among them, the Eurotium rubrum SH58 fastest growth, stable biological characteristics.Therefore, choose the strain for the further study.2.the Eurotium rubrum SH58 the basis of studying the biological characteristics, this research studied the esterase.With e. rubrum SH58 m RNA for materials, synthesis c DNA by reverse transcription PCR, we cloned a Erlipase esterase gene;And the prokaryotic expression and purification of fusion protein.Through to the esters degradation activity determination, proved that the prokaryotic expression of gene product has the high activity of esterase.The above results show that e. rubrum SH58 oil degradation characteristics, and the characteristic that is related to its Erlipase gene expression.Research not only provides the Marine oil pollution control biological materials are available, and also for the microbial genetic improvement of environmental pollution treatment and relevant laid a foundation.