Identification and Functional Characterization of the FATP1 Gene from Mud Crab, Scylla paramamosain.

In mammals, fatty acid transport protein 1 (FATP1) plays important roles in cellular uptake and activation of long-chain fatty acid (LCFA), especially in processes of transportation, oxidation and triacylglycerol synthesis. However, the role of FATP1 in invertebrates, especially decapod crustaceans, is still poorly understood. In this study, the cDNA of a FATP1 gene from a decapod crustacean, mud crab Scylla paramamosain, was cloned and functionally characterized. The FATP1 gene encoded a polypeptide consisting of 643 amino acids that exhibits all the typical features of the FATP family and shares high homology with the other FATP orthologs of crustaceans. The relative mRNA expression levels of FATP1 were observed to be higher in metabolically active tissues such as hepatopancreas, stomach and gill than in other crab parts. Knockdown of the FATP1 mRNA in vivo significantly reduced triacylglycerols and total lipid levels in the hepatopancreas, accompanied by an increase in the expression of genes related to fatty acid transportation, allocation and hydrolysis, including long-chain acyl-CoA synthetase 3/4 (ACSL3/4) and carnitine palmitoyl transferase 1 (CPT1), and a decrease in the expression of genes related to fatty acid synthesis such as acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS) in the hepatopancreas. Furthermore, increased dietary n-3 long-chain polyunsaturated fatty acid (LC-PUFA) levels resulted in the up-regulation of the FATP1 expression in the hepatopancreas, accompanied by an increase in LC-PUFA content, especially eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), in both polar (PLs) and neutral lipids (NLs) in the hepatopancreas and muscles of crabs. These findings suggested that the FATP1 gene identified in S. paramamosain might play important roles in regulating long-chain fatty acid metabolism and deposition in crustaceans.

Dietary fucoidan extracted from macroalgae Saccharina japonica alleviate the hepatic lipid accumulation of black seabream (Acanthopagrus schlegelii).

The use of an artificial diet often leads to the increase of risk factors for the development of liver diseases, such as hepatic lipid accumulation (HLA) in commercially cultured fish species. Our previous study showed that dietary Saccharina japonica could effectively alleviate HLA in black seabream (Acanthopagrus schlegelii), which may be linked predominantly to S. japonica fucoidan. Thus, a 56d nutritional trial was designed to investigate the effects of dietary fucoidan (CTRL, 0 g kg-1; ASJ1, 0.75 g kg-1; ASJ2, 3.00 g kg-1) on growth performance, fillets nutritional values, and HLA of black seabream. Results showed that dietary fucoidan significantly improved the growth and the contents of n-3 polyunsaturated fatty acids (n-3PUFA) in fillets of black seabream. Moreover, dietary fucoidan improved HLA-related parameters, including reducing serum and liver lipid contents and the activity of aminotransferase. Meanwhile, histological analysis showed that dietary fucoidan reduced the area of hepatic lipid droplets in black seabream (P < 0.05). In addition, the transcriptomic analysis of differentially expressed gene (DEG) showed that all DEG in fatty acid metabolism, primary bile acid biosynthesis, and fatty acid biosynthesis were down-regulated, and all DEG in the regulation of autophagy were up-regulated in the ASJ1 group compared with CTRL group. Moreover, the metabolomic analysis of differentially expressed metabolite (DEM) found that lipid metabolism was the main type of KEGG pathway altered by fucoidan supplementation. Furthermore, the combined transcriptomic and metabolomic analysis found that dietary fucoidan mainly modified the lipid metabolic pathway of primary bile acid biosynthesis, glycerophospholipid metabolism, and arachidonic acid metabolism in the liver. In general, dietary fucoidan effectively alleviated HLA of black seabream, and the underlying mechanism may be ascribed to promoting the autophagy and inhibiting the synthesis of lipids and bile acids.

Identification of miR-145 as a Key Regulator Involved in LC-PUFA Biosynthesis by Targeting hnf4α in the Marine Teleost Siganus canaliculatus.

Fish, particularly marine species, are considered as the major source of long-chain polyunsaturated fatty acids (LC-PUFA) in the human diet. The extent to which fish can synthesize LC-PUFA varies with species and is regulated by dietary fatty acids and ambient salinity. Therefore, to enable fish to produce more LC-PUFA, comprehending the mechanisms underlying the regulation of LC-PUFA biosynthesis is necessary. Here, the regulatory roles of miR-145 were investigated in the marine teleost rabbitfish Siganus canaliculatus. The hepatic abundance of miR-145 was lower in rabbitfish reared in low salinity (10 ppt) in comparison with that of those cultured in seawater (32 ppt), while the opposite pattern was observed for the transcripts of the transcription factor hepatocyte nuclear factor 4 alpha (Hnf4α), known to affect rabbitfish LC-PUFA biosynthesis. Rabbitfish hnf4α was identified as a target of miR-145 by luciferase reporter assays, and overexpression of miR-145 in the S. canaliculatus hepatocyte line (SCHL) markedly reduced the expression of Hnf4α and its target genes involved in LC-PUFA biosynthesis, namely, Δ4 fads2, Δ6Δ5 fads2, and elovl5. The opposite pattern was observed when miR-145 was knocked down in SCHL cells, with these effects being attenuated by subsequent hnf4α knockdown. Moreover, increasing endogenous Hnf4α by the knockdown of miR-145 increased the expression of LC-PUFA biosynthesis genes and enhanced the synthesis of LC-PUFA in both SCHL cells and rabbitfish in vivo. This is the first report to identify miR-145 as a key effector of LC-PUFA biosynthesis by targeting hnf4α, providing a novel insight into the mechanisms of the regulation of LC-PUFA biosynthesis in vertebrates.

miR-26a mediates LC-PUFA biosynthesis by targeting the Lxrα-Srebp1 pathway in the marine teleost Siganus canaliculatus.

MicroRNAs have been recently shown to be important regulators of lipid metabolism. However, the mechanisms of microRNA-mediated regulation of long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis in vertebrates remain largely unknown. Herein, we for the first time addressed the role of miR-26a in LC-PUFA biosynthesis in the marine rabbitfish Siganus canaliculatus The results showed that miR-26a was significantly down-regulated in liver of rabbitfish reared in brackish water and in S. canaliculatus hepatocyte line (SCHL) incubated with the LC-PUFA precursor α-linolenic acid, suggesting that miR-26a may be involved in LC-PUFA biosynthesis because of its abundance being regulated by factors affecting LC-PUFA biosynthesis. Opposite patterns were observed in the expression of liver X receptor α (lxrα) and sterol regulatory element-binding protein-1 (srebp1), as well as the LC-PUFA biosynthesis-related genes (Δ4 fads2, Δ6Δ5 fads2, and elovl5) in SCHL cells incubated with α-linolenic acid. Luciferase reporter assays revealed rabbitfish lxrα as a target of miR-26a, and overexpression of miR-26a in SCHL cells markedly reduced protein levels of Lxrα, Srebp1, and Δ6Δ5 Fads2 induced by the agonist T0901317. Moreover, increasing endogenous Lxrα by knockdown of miR-26a facilitated Srebp1 activation and concomitant increased expression of genes involved in LC-PUFA biosynthesis and consequently promoted LC-PUFA biosynthesis both in vitro and in vivo These results indicate a critical role of miR-26a in regulating LC-PUFA biosynthesis through targeting the Lxrα-Srebp1 pathway and provide new insights into the regulatory network controlling LC-PUFA biosynthesis and accumulation in vertebrates.

Adsorption isotherm, kinetics simulation and breakthrough analysis of 5-hydroxymethylfurfural adsorption/desorption behavior of a novel polar-modified post-cross-linked poly (divinylbenzene-co-ethyleneglycoldimethacrylate) resin.

A polar modified post-cross-linked poly (divinylbenzene-co-ethyleneglycol-dimethacrylate) (PCL-PDE) resin was synthesized by suspension polymerization of ethylene glycol dimethacrylate (EGDMA) and divinylbenzene (DVB), and a post-cross-linked reaction. After characterization, the adsorption behaviors of 5-hydroxymethylfurfural (5-HMF) on PCL-PDE resin were determined in comparison with the starting copolymers PDE resin. The equilibrium adsorption capacity of 5-HMF on PCL-PDE resin was much larger than PDE resin and the increase rate was greater than 52.6%. The equilibrium data of 5-HMF onto PCL-PDE resin were found to be better fitted by the Langmuir isotherm model. The kinetic data shows that the adsorption reached equilibrium in a short time (less than 20 min) can be fitted by the pore diffusion model (PDM) at various operating conditions. The effective pore diffusion coefficient was dependent upon adsorption temperature, and were 6.706 × 10-10, 8.958 × 10-10, 1.136 × 10-9 and 1.429 × 10-9 m2 s-1 at 288, 298, 308 and 318 K, respectively. Furthermore, the effects of feed flow rate (Qf = 0.6, 1.5, 3.0 and 6.0 mL min-1) and initial 5-HMF concentration (cf = 0.52, 1.02, 2.00 and 4.96 g L-1) on the adsorption were investigated systematically. Besides, a general rate model (GRM) was used to predict adsorption breakthrough curves of 5-HMF. The simulation results are highly consistent with the experimental data, indicating that the GRM can successfully simulate this process. In the desorption process, the desorption capacity reaches 99.6% of adsorbed capacity, suggesting that the PCL-PDE resin exhibited good reusability. Therefore, it could be suggested that the PCL-PDE resin has a potential application in the separation and purification of 5-HMF.

Enhanced Enzymatic Hydrolysis and Lignin Extraction of Wheat Straw by Triethylbenzyl Ammonium Chloride/Lactic Acid-Based Deep Eutectic Solvent Pretreatment.

Efficient and feasible pretreatment of lignocellulosic biomass waste is an important prerequisite step to promote subsequent enzymatic hydrolysis and enhance the economics of biofuels production. This study focuses on the pretreatment of wheat straw (WS) with triethylbenzyl ammonium chloride/lactic acid (TEBAC/LA)-based deep eutectic solvents to enhance biomass fractionation and lignin extraction. Effects of pretreatment time, temperature, and TEBAC/LA molar ratio on pretreatment were evaluated systematically. Results suggested that 89.06 ± 1.05% of cellulose and 71.00 ± 1.03% of xylan were hydrolyzed with enzyme loadings of 35 FPU cellulase and 82 CBU ß-glucosidase (per gram of dry biomass) after pretreatment by TEBAC/LA (1:9) at 373 K for 10 h. A total monosaccharide yield of 0.550 g/g WS (91.27% of the theoretical yield) was achieved with 79.73 ± 0.93% of lignin removal. Furthermore, the 1H-13C two-dimensional heteronuclear single quantum correlation (2D-HSQC) NMR spectroscopy showed that the regenerated lignin (75.69 ± 1.32% purity) was mainly composed of the syringyl units and the guaiacyl units. Overall, the results in this study provide an effective and facile pretreatment method for lignocellulosic biomass waste to enhance enzymatic hydrolysis saccharification.