A new member of the CYP116B subfamily-P450LaMO-was discovered in Labrenzia aggregata by genomic data mining. It was successfully overexpressed in Escherichia coli, purified, and subsequently characterized spectroscopically, and its catalytic properties were assessed. Substrate profiling of the P450LaMO revealed that it was a versatile catalyst, exhibiting hydroxylation and epoxidation activities as well as O-dealkylation and asymmetric sulfoxidation activities. Diverse compounds, including alkylbenzenes, aromatic bicyclic molecules, and terpenoids, were shown to be hydroxylated by P450LaMO. Such diverse catalytic activities are uncommon for the bacterial P450s, and the P450LaMO-mediated stereoselective hydroxylation of inactivated C-H bonds-ubiquitous and relatively unreactive in organic molecules-is particularly unusual. The self-sufficient nature of P450LaMO, coupled with its broad substrate range, highlights it as an ideal template for directed evolution towards various applications.

Abstract. Extraction of Omega-3 fatty acid from Catfish using enzymatic hydrolysis techniques are expected to be more economically possible techniques due to the uses of enzyme with the characteristic of reusable, environmental friendly and less energy required for large-scale production. Malaysian like to consume Catfish in their daily diet. Thus, it is very popular among farmers in Malaysia. Omega-3 polyunsaturated fatty acid have many benefit to human health. Based on research, EPA and DHA can reduce the risk of heart disease by regulate the blood coagulation. The main purpose of this research is to extract the Omega-3 polyunsaturated fatty acid by using enzymatic hydrolysis method. Enzymatic extraction technique by using Alcalase was used to extract the Catfish oil. Then, lipase was used to concentrate the omega-3 polyunsaturated fatty acid. Peroxide value and acid value were used to determine the Catfish oil quality. Optimization using DOE verify that the highest percentage of oil yields at 51°C, 0.91% enzyme concentration and 155 minute incubation time. Finally, there are two types of omega-3 polyunsaturated fatty acid can be found in Catfish oil which is EPA and ALA.

Four types of O-methylated substrates were designed as probes for the detection of fingerprints of Type IV P450s.

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