A comparative study was done on the production of different sterol esters using a stirred tank batch reactor (STBR) and packed bed reactor (PBR) using Thermomyces lanuginosus (Lipozyme TLIM) enzyme, a commercially immobilized lipase. Different oils were used as the sources of particular fatty acids, e.g., fish oil for n-3 polyunsaturated fatty acids (n-3 PUFA), linseed oil for alpha linolenic acid (ALnA) and mustard oil for erucic acid. Reaction parameters, such as substrate molar ratio, reaction temperature and enzyme concentration, were standardized in the STBR and maintained in the PBR. To provide equal time of residence between the substrate and enzyme in both the reactors for the same amount of substrates, the substrate flow rate in the PBR was maintained at 0.27 ml/min. Thin layer chromatography was used to monitor the reaction, and column chromatography was used to determine the product yields. Fatty acid compositions of the esters were determined by gas chromatography. The study showed that the packed bed bioreactor was more efficient than the batch reactor in sterol-ester synthesis with less migration of acyl groups.

Structured lipids were prepared from mustard oil by enzymatic acidolysis reaction with capric acid (C10) using lipase enzyme TLIM from Thermomyces lanuginosus as biocatalyst. Parameters such as substrate molar ratio, enzyme concentration, reaction temperature, stirring speed and time of maximum incorporation, were studied for the optimization of the reaction. The optimized set of process conditions was predicted by response surface methodology (RSM) and genetic algorithm (GA). The robustness of GA and RSM was evaluated using regression coefficient and p value. The R(2) found out by GA was 0.996 while from RSM was 0.973. The results proved that GA models have better performance than RSM models. From the result, it could be concluded that optimal conditions for synthesis of capric acid rich mustard oil were: Temperature = 39.5 °C ; time = 21.1 hr; Substrate ratio = 3.5; Enzyme content = 8.8%; Speed = 570.8 rpm.

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