A computational search for lipases that can preferentially hydrolyze long-chain omega-3 fatty acids from fish oil triacylglycerols

Consumption of long-chain omega-3 fatty acids is known to decrease the risk of major cardiovascular events. Lipases, a class of triacylglycerol hydrolases, have been extensively tested to concentrate omega-3 fatty acids from fish oils, under mild enzymatic conditions. However, no lipases with preference for omega-3 fatty acids selectivity have yet been discovered or developed. In this study we performed an exhaustive computational study of substrate-lipase interactions by docking, both covalent and non-covalent, for 38 lipases with a large number of structured triacylglycerols containing omega-3 fatty acids. We identified some lipases that have potential to preferentially hydrolyze omega-3 fatty acids from structured triacylglycerols. However omega-3 fatty acid preferences were found to be modest. Our study provides an explanation for absence of reports of lipases with omega-3 fatty acid hydrolyzing ability and suggests methods for developing these selective lipases.

Co-encapsulation and characterisation of omega-3 fatty acids and probiotic bacteria in whey protein isolate-gum Arabic complex coacervates

Omega-3 fatty acids and probiotic bacteria were co-encapsulated in a single whey protein isolate (WPI)-gum Arabic (GA) complex coacervate microcapsule. Tuna oil (O) and Lactobacillus casei 431 (P) were used as models of omega-3 and probiotic bacteria, respectively. The co-microcapsules (WPI-P-O-GA) and L.casei containing microcapsules (WPI-P-GA) were converted into powder by using spray and freeze drying. The viability of L.casei was significantly higher in WPI-P-O-GA co-microcapsules than in WPI-P-GA. The oxidative stability of tuna oil was significantly higher in spray dried co-capsules than in freeze dried ones.

Exploring omega-3 fatty acids, enzymes and biodiesel producing thraustochytrids from Australian and Indian marine biodiversity

The marine environment harbours a vast diversity of microorganisms, many of which are unique, and have potential to produce commercially useful materials. Therefore, marine biodiversity from Australian and Indian habitat has been explored to produce novel bioactives, and enzymes. Among these, thraustochytrids collected from Indian habitats were shown to be rich in saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs), together constituting 51-76 % of total fatty acids (TFA). Indian and Australian thraustochytrids occupy separate positions in the dendrogram, showing significant differences exist in the fatty acid profiles in these two sets of thraustochytrid strains. In general, Australian strains had a higher docosahexaenoic acid (DHA) content than Indian strains with DHA at 17-31 % of TFA. A range of enzyme activities were observed in the strains, with Australian strains showing overall higher levels of enzyme activity, with the exception of one Indian strain (DBTIOC-1). Comparative analysis of the fatty acid profile of 34 strains revealed that Indian thraustochytrids are more suitable for biodiesel production since these strains have higher fatty acids content for biodiesel (FAB, 76 %) production than Australian thraustochytrids, while the Australian strains are more suitable for omega-3 (40 %) production.

Effect of dietary saturated and monounsaturated fatty acids in juvenile barramundi Lates calcarifer

Barramundi (Lates calcarifer), a catadromous teleost of commercial interest, perform well when fed a wide range of dietary oils. However, the range of alternative oils now being explored is typically rich in saturated and monounsaturated fatty acids (SFA and MUFA). In this study, the response of juvenile barramundi (47.0 g per fish initial weight) fed isolipidic and isoenergetic diets with 82 g kg−1 added oil was tested. The experimental test diets had a 2 : 1 or 1 : 2 ratio of SFA to MUFA (SFA-D and MUFA-D, respectively) compared to a control diet (CTRL-D) fed for 8 weeks. The diets containing mostly olive oil (dietary MUFA-D) and mostly refined palm oil (dietary SFA-D) did not impact the growth performance or feed utilization parameters of the barramundi. The in vivo beta-oxidation activity was consistent with the dietary fatty acid composition, with the most dominant FA being heavily beta-oxidized. Together, the in vivo whole-body mass balance of fatty acids showed that n-3 long-chain polyunsaturated fatty acids (LC-PUFA) were most efficiently utilized in the SFA-D- and MUFA-D-fed fish. This study provides evidence that additional dietary MUFA and SFA are suitable lipid classes for juvenile barramundi and they are both equally efficient at sparing LC-PUFA from an oxidative fate.

Omega-3 fatty acids produced from microalgae

The applications of Omega-3 fatty acids for human health are rapidly expanding, which necessitates exploring alternative sources to fish. Many marine microorganisms across different kingdoms exhibit the ability to store a significant oil content, however are difficult to cultivate. Out of all marine microbes, thraustochytrids are considered a good source for the production of high value compounds such as polyunsaturated fatty acids (PUFAs). Optimization of culture conditions will be helpful in further enhancing cellular lipid content to suit fatty acid synthesis. This chapter describes some recent advances in the development of marine microbes for fatty acid production with a special emphasis upon thraustochytrids for biotechnological applications, focussing particularly on methods to enhanced docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) production.

Selective enrichment of omega-3 fatty acids in oils by phospholipase A1

Omega fatty acids are recognized as key nutrients for healthier ageing. Lipases are used to release ω-3 fatty acids from oils for preparing enriched ω-3 fatty acid supplements. However, use of lipases in enrichment of ω-3 fatty acids is limited due to their insufficient specificity for ω-3 fatty acids. In this study use of phospholipase A1 (PLA1), which possesses both sn-1 specific activity on phospholipids and lipase activity, was explored for hydrolysis of ω-3 fatty acids from anchovy oil. Substrate specificity of PLA1 from Thermomyces lenuginosus was initially tested with synthetic p-nitrophenyl esters along with a lipase from Bacillus subtilis (BSL), as a lipase control. Gas chromatographic characterization of the hydrolysate obtained upon treatment of anchovy oil with these enzymes indicated a selective retention of ω-3 fatty acids in the triglyceride fraction by PLA1 and not by BSL. 13C NMR spectroscopy based position analysis of fatty acids in enzyme treated and untreated samples indicated that PLA1 preferably retained ω-3 fatty acids in oil, while saturated fatty acids were hydrolysed irrespective of their position. Hydrolysis of structured triglyceride,1,3-dioleoyl-2-palmitoylglycerol, suggested that both the enzymes hydrolyse the fatty acids at both the positions. The observed discrimination against ω-3 fatty acids by PLA1 appears to be due to its fatty acid selectivity rather than positional specificity. These studies suggest that PLA1 could be used as a potential enzyme for selective concentrationof ω-3 fatty acids.

Relationships of nicotianamine and other amino acids with nickel, zinc and iron in Thlaspi hyperaccumulators

Experimental evidence suggests that nicotianamine (NA) is involved in the complexation of metal ions in some metal-hyperaccumulating plants. Closely-related nickel (Ni)- and zinc (Zn)-hyperaccumulating species were studied to determine whether a correlation exists between the Ni and Zn concentrations and NA in foliar tissues. A liquid chromatography–mass spectrometry (LC-MS) procedure was developed to quantify the NA and amino acid contents using the derivatizing agent 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. A strong correlation emerged between Ni and NA, but not between Zn and NA. Concentrations of NA and l-histidine (His) also increased in response to higher Ni concentrations in the hydroponic solution supplied to a serpentine population of Thlaspi caerulescens. An inversely proportional correlation was found between the iron (Fe) and Ni concentrations in the leaves. Correlations were also found between Zn and asparagine. The results obtained in this study suggest that NA is involved in hyperaccumulation of Ni but not Zn. The inverse proportionality between the Ni and Fe concentrations in the leaf may suggest that Ni and Fe compete for complexation to NA.