Response surface methodology for optimising the culture conditions for eicosapentaenoic acid production by marine bacteria

Polyunsaturated fatty acids (PUFAs), especially eicosapentaenoic acid (EPA), are increasingly attracting scientific attention owing to their significant health-promoting role in the human body. However, the human body lacks the ability to produce them in vivo. The limitations associated with the current sources of ω-3 fatty acids from animal and plant sources have led to increased interest in microbial production. Bacterial isolate 717 was identified as a potential high EPA producer. As an important step in the process development of the microbial PUFA production, the culture conditions at the bioreactor scale were optimised for the isolate 717 using a response surface methodology exploring the significant effect of temperature, pH and dissolved oxygen and the interaction between them on the EPA production. This optimisation strategy led to a significant increase in the amount of EPA produced by the isolate under investigation, where the amount of EPA increased from 9 mg/g biomass (33 mg/l representing 7.6 % of the total fatty acids) to 45 mg/g (350 mg/l representing 25 % of the total fatty acids). To avoid additional costs associated with extreme cooling at large scale, a temperature shock experiment was carried out reducing the overall cooling time from the whole cultivation process to 4 h only prior to harvest. The ability of the organism to produce EPA under the complete absence of oxygen was tested revealing that oxygen is not critically required for the biosynthesis of EPA but the production improved in the presence of oxygen. The stability of the produced oil and the complete absence of heavy metals in the bacterial biomass are considered as an additional benefit of bacterial EPA compared to other sources of PUFA. To our knowledge this is the first report of a bacterial isolate producing EPA with such high yields making the large-scale manufacture much more economically viable.