Fatty Acid Methyl Ester Analysis of Olive Oil Degraded by Pseudomonas fluorescens and Enzymatic Characterisation of the Lipase

Vegetable oils are harmful to the environment, like petroleum oils they produce similar environmental effects. Hence, it is a known fact that the olive oil causes the production of large quantity of olive mill waste (OMWs) as by-products to the environment. These by-products are harmful to the environment, both  terrestrial  and  aquatic.  Bacteria  are  very  useful  in  protecting  the  environment.  The  lipolytic activities  of  physiologically  diverse  bacteria  have  great  potential  to  degrade  oil  spills  in  the environment. Fatty  acids  are  the  major  components  of  lipids;  and  the  physical,  chemical  and physiological properties of a lipid class depend primarily on its fatty acid composition.There is need for extensive characterisation of the bacterium lipase for the treatment of vegetable oil-polluted sites. This work was carried out to preliminarily characterise the lipase of Pseudomonas fluoresens and to check  the  fatty  acid  composition  of  olive  oil.Gas  chromatography  method  can  be  used  for  the identification of microbiological degraded fatty acids in vegetable oils as methyl ester.Pseudomonas fluoresenswas screened for lipase production using standard methods. Temperature, pH, ion concentration (NaNO3and MgSO4), enzyme concentration, nitrogen concentration, substrate concentration,  time  course  and  agitation  speed  were  optimised  for  the  lipase  activity  as  well  as growth.Crude enzyme of Pseudomonas fluoresenshad the highest lipase activity and growth of 0.8 U/mL and 1.418 mg/mL respectively at room temperature, but when production was optimised higher activity 0.9 U/mL was seen  in the use of glucose as substrate. Agitation with the speed used did not support lipase  production  but  supported  growth  (1.998  mg/mL)  at  agitation  speed  of  100  rpm. Olive  oil degraded by  lipase of Pseudomonas fluorescens grown in two different  Mineral Salt Medium, was studied over 25 days. Olive oil was analyzed for fatty acids commonly present in olive oils which are Myristic,  Palmitic,  Stearic,  Oleic,  Linoleic,  Linolenic,  Behenic  and  Lignoceric,  which  have  specific carbon number and their values in approximate percentage are C14:0 (0.4), C16:0 (14.0), C18:0 (5.5), C18:1  (76.4),  C18:2  (3.4),  C18:3  (0.1),  C22:0  (0.1)  and  C24:0  (0.1)  respectively.  Oleic  acid percentage is high in olive oil which contained considerable amount of 76.4%.The reduction of fatty acid by Pseudomonas fluorescens after 20 days was 8.2% in the media used. Lipase from Pseudomonas fluorescens had potential for degradation of fatty waste. It could therefore be employed in environmental cleanup of oil spill site. Lipase production is generally influenced by the type and concentration of carbon and nitrogen sources, the culture pH, the growth temperature and the dissolved oxygen  concentration [1].Hence  monitoring the result of degradation patterns of the vegetable oil using fatty acid methyl ester analysis, by Pseudomonas fluorescens, showed that this organism has great potential for fatty waste degradation and as an extension in environmental clean up of oil spill site.

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