While optimizing the lipase production, oil concentrations (tributyrin oil) from 5-25% v/v were taken, and it was observed that enzyme activity was decreased with an increase in oil concentration.
Why? Thank you in Advance.
Hi there,
Is it less protein produced or less activity detected? If the latter the lipase might not be stable when oïl concentration becomes too high (ie. when the hydrophobicity of the media increases)...
High oil-concentration will lead to bigger but less drops of oil, which have a higher volume/surface-ratio. Since lipase is only working on the oil/water-interface, there is less surface for more volume of oil for lipase to work on - Your activity assay probably does not take this into account.
Adding an emulgating agent might help.
Lipases act preferably at oil water interface .Increased oil or triglyceride in the production media will not increase the lipase activity .The availability of the substrate can be enhanced by addition of any emulsifying agents such as Poly vinyl alcohol ,Gum acacia,Gum arabic etc . Availability of substrate and structural features of a enzyme (Lipase ) are the two important factors which may probably inhibiting the enzyme activity .
25% of tributyrin is too high. Tributyrin will enhance the lipase production at the same time excess of Tributyrin will inhibit the enzyme just like substrate feed back inhibition.
Thank you all for the suggestions and a nice conversation on this topic.
# As Ravi Mohan said, I used the emulsifying agent gum acacia in my production medium and It was seen that 5% oil was optimum for the lipase production than more than that. which denotes minimum substrate (oil) is inducible than the higher concentration for lipase production.
# Dr. Jai Ghosh, The lipase was produced by SmF technique using tributyrin oil as substrates by Pseudomonas sp.
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