**2.1 Transformation of soybean oil to fatty acid vesicles**

In our study, the selected microbes, here in, *Candida* sp. grown in the medium containing carbohydrates, peptone, mineral salts and soybean oil at different percentages (w/v) for the period of more than 5-9 days at 37C under agitation at 180-200rpm, we observed the slow transformation of these oils to micelle and then to vesicles. Though different concentrations of oils were attempted, only at higher concentrations (>5%) we observed the final transformed product as fatty acid vesicles. In addition to these observations, the physicochemical analysis of the medium on different days clearly illustrates the steps involved in the process of transformations. We found, the microbes, in order to utilize the substrates released the enzymes and surface-active agents externally. Analysis of enzyme release suggests, lipase was released from 12 hours onwards at the rate of 3.5 U/ml. The produced lipase interacted with the oil present in the external aqueous medium and hydrolyzes it into fatty acids with in 48 hours. In the meantime, the organism produces surface-active agents (33 2 mN/m), which further starts interacting with the hydrolyzed fatty acids. Thus, a simultaneous reaction takes place between the fatty acids released upon enzymatic hydrolysis and the surface-active agents produced. Figure 1 illustrates the vesicle formed in the growth medium.

Fig. 1. Fatty acid vesicle from soybean oil.

digest of animal tissues; 1g yeast extract; 0.1g ferric citrate; 19.45g sodium chloride; 8.8g magnesium chloride; 3.24g sodium sulphate; 1.80g calcium chloride; 0.55g potassium chloride; 0.16g sodium bicarbonate; 0.08g potassium bromide; 0.034g; strontium chloride; 0.022 boric acid; 0.004 sodium silicate; 0.0024g sodium fluorate; 0.0016g ammonium nitrate;

The selective microorganisms; marine *Bacillus* sps and clinical strain *Candida albicans* are cultured in the selective medium with triglycerides at different volumes (0.5, 1.0, 1.5, 2.0, 2.5, 4, 6, 8 and 10% w/v) and at cell concentration of 1X 105 cells per ml and incubated for the period of 5-10 days under shaking/ agitation at 180-200 rpm. Followed by inoculation, observations on cell growth, pH profile, hydrolytic enzyme production, fatty acids and glycerol release, surface-active agent production, transformation of oil, micelle formation, nano vesicle to macrosize supramolecular structures were made. The descriptions on

In our study, the selected microbes, here in, *Candida* sp. grown in the medium containing carbohydrates, peptone, mineral salts and soybean oil at different percentages (w/v) for the period of more than 5-9 days at 37C under agitation at 180-200rpm, we observed the slow transformation of these oils to micelle and then to vesicles. Though different concentrations of oils were attempted, only at higher concentrations (>5%) we observed the final transformed product as fatty acid vesicles. In addition to these observations, the physicochemical analysis of the medium on different days clearly illustrates the steps involved in the process of transformations. We found, the microbes, in order to utilize the substrates released the enzymes and surface-active agents externally. Analysis of enzyme release suggests, lipase was released from 12 hours onwards at the rate of 3.5 U/ml. The produced lipase interacted with the oil present in the external aqueous medium and hydrolyzes it into fatty acids with in 48 hours. In the meantime, the organism produces surface-active agents (33 2 mN/m), which further starts interacting with the hydrolyzed fatty acids. Thus, a simultaneous reaction takes place between the fatty acids released upon enzymatic hydrolysis and the surface-active agents produced. Figure 1 illustrates the vesicle formed in

various supramolecular structures are briefly explained below:

**2.1 Transformation of soybean oil to fatty acid vesicles** 

0.008g disodium phosphate.

the growth medium.

Fig. 1. Fatty acid vesicle from soybean oil.

Though, we couldn't observe the complete hydrolysis of oil with that of the lipase produced, however, the unhydrolyzed oil further solubilized by the released surface-active agents and interact with the already formed vesicles and helps in the assembly of vesicles. In addition, in the case of the chosen oils, the presence of lecithin increases the supramolecular selfassembly results with the increased gellation with multilamellar vesicles. The release of amino acids during the growth of the microorganism may also involved in the transformation of vegetable oil to vesicles.
