Conjugated linoleic acid (CLA) and polyunsaturated fatty acids in muscle lipids of lambs from the Patagonian area of Argentina

The concentrations of fatty acids were measured in total lipids, triacyglycerol and phospholipid fractions of intramuscular fat (IMF) from the Longissimus dorsi (LD) muscle of 10 lambs reared to approximately 30kg live weight on natural pasture with their dams. Fatty acid composition was also measured in 25 (five of each) Semitendinosus (ST), Semimembranosus (SM), Rectus femoris (RF), Gluteus (GLU) and Tensor fascia latea (TFL) muscles. Intramuscular fat percentages were similar for all muscles. Aspects of the fatty-acid patterns of relevance to human nutrition tended to favor the leg muscles with lower saturated fatty acids (SFA %), n-6/n-3 fatty acid ratios (p<0.01) and higher concentrations of the conjugated linoleic acid (CLA) (p<0.05). The estimated fatty acid concentrations (mg/100g of meat) showed higher contribution of arachidonic (C20:4 n-6), eicosapentanoic (C20:5 n-3), docosapentanoic (C22:5 n-3) and docosahexanoic (C22:6 n-3) acids in leg compared to LD lipids.     

Antioxidant activities of phenolic rich fractions (PRFs) obtained from black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes

The antioxidant activities of phenolic rich fractions (PRFs) from crude methanolic extract (CME), and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes were investigated. The total phenolic compounds were found to be higher in white mahlab than black mahlab seedcakes. The antioxidant activity determined by the DPPH method revealed that black mahlab PRFs had the highest antioxidant activity, compared to white mahlab fractions. The presence of antioxidants in the two mahlab PRFs reduced the oxidation of β-carotene by hydroperoxides from these extracts/fractions. The effect of the two mahlab PRFs on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The CME performed better antioxidant activity in inhibiting the formation of both primary and secondary oxidation products. The qualitative and quantitative characterisation of phenolic compounds was carried out by HPLC/DAD.     

Heterotrophic growth and nutritional aspects of the diatom Cyclotella cryptica (Bacillariophyceae): Effect of some environmental factors

To investigate the nutritional value of the diatom Cyclotella cryptica (Reimann, Lewin, and Guillard) as an alternative feed for use in the aquaculture industry, the heterotrophic growth characteristics and resultant fatty acid profile of the microalga were studied when cultivated under a variety of controlled salinity and temperature conditions. In addition, the effects of pH on the growth characteristics were also studied. The maximum specific growth rate was affected by initial pH and cultivation temperature, but not by salinity. The optimal pH and temperature ranges for growth were 7.2 to 8.1 and 22.5 to 25.0 °C, respectively. Lipid accumulation and the fatty acid composition were also affected by cultivation temperature and salinity. The optimal temperature range and salinity level for lipid accumulation were 18.0 to 25.0 °C and 11.2 psu, respectively. In all cases the fatty acid distribution was similar, with the most abundant fatty acids being palmitic acid (16:0), palmitoleic acid (16:1 n-7), stearidonic acid (18:4 n-3, SDA), eicosapentaenoic acid (20:5 n-3, EPA), and decosahexaenoic acid (22:6 n-3, DHA).     

Antioxidant activity and phenolic content of phenolic rich fractions obtained from black cumin (Nigella sativa) seedcake

The antioxidant activities of crude methanolic extract (CME) and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black cumin seedcake were investigated. DPPH radical scavenging activity, β-carotene–linoleate bleaching, and inhibition of corn oil oxidation were used to evaluate the antioxidant capacity. The total phenolics were found to be 78.8, 27.8, 32.1 and 12.1 mg gallic acid equivalents (GAE)/g in EAF, CME, WF and HF, respectively. The CME and EAF exhibited the highest DPPH followed by WF and HF. The extract/fractions showed high effect on reducing the oxidation of β-carotene. The effect of extract/fractions on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The oil peroxide and anisidine values were generally lower with addition of PRFs in comparison to a control. The predominant phenolic compounds identified by HPLC–DAD in CME and WF of black cumin seedcake were hydroxybenzoic, syringic and p-cumaric acids.     

Taxonomic characterization and metabolic analysis of the Halomonas sp. KM-1, a highly bioplastic poly(3-hydroxybutyrate)-producing bacterium

In a brief previous report, the gram-negative moderately halophilic bacterium, Halomonas sp. KM-1, that was isolated in our laboratory was shown to produce the bioplastic, poly(3-hydroxybutyrate) (PHB), using biodiesel waste glycerol (Kawata and Aiba, Biosci. Biotechnol. Biochem., 74, 175-177, 2010). Here, we further characterized this KM-1 strain and compared it to other Halomonas strains. Strain KM-1 was subjected to a polyphasic taxonomic study. Strain KM-1 was rod-shaped and formed colonies on a plate that were cream-beige in color, smooth, opaque, and circular with entire edges. KM-1 grew under environmental conditions of 0.1%-10% (w/v) NaCl, pH 6.5-10.5 and at temperatures between 10°C and 45°C. The G+C content of strain KM-1 was 63.9 mol%. Of the 16 Halomonas strains examined in this study, the strain KM-1 exhibited the highest production of PHB (63.6%, w/v) in SOT medium supplemented with 10% glycerol, 10.0 g/L sodium nitrate and 2.0 g/L dipotassium hydrogen phosphate. The intracellular structures within which PHB accumulated had the appearance of intracellular granules with a diameter of approximately 0.5 μm, as assessed by electron microscopy. The intra- and extra-cellular metabolites of strain KM-1 were analyzed by capillary electrophoresis mass spectrometry. In spite of the high amount of PHB stored intra-cellularly, as possible precursors for PHB only a small quantity of 3-hydroxybutyric acid and acetyl CoA, and no quantity of 3-hydroxybutyl CoA, acetoacetyl CoA and acetoacetate were detected either intra- or extra-cellularly, suggesting highly efficient conversion of these precursors to PHB.     

Production of triacylglycerol and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by the toluene-degrading bacterium Rhodococcus aetherivorans IAR1

Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) produced by various bacteria has been intensively investigated as a promising biodegradable plastic, but required a supply of an expensive precursor as a secondary carbon source for its production. In a previous study, we identified a new bacterial strain, Rhodococcus aetherivorans IAR1, which synthesizes PHBV from toluene without the supply of a precursor. Toluene is the volatile organic compound most abundantly emitted to the environment. In the present paper, we show that R. aetherivorans IAR1 produces triacylglycerols (TAGs) simultaneously with PHBV. Both PHBV and TAGs were synthesized before the nitrogen source is completely exhausted. The cellular content of PHBV reached 10% of cell dry weight (CDW) and its synthesis ceased even during intermittent supply of toluene. However, accumulation of TAGs continued during cultivation and their cellular content reached 24% of CDW at the end of cultivation. Cerulenin inhibited TAG production and increased PHBV cellular content up to 30% of CDW. The mole fraction of 3-hydroxyvalerate (3HV) in PHBV produced from toluene increased from 60% to 80% during its accumulation. Fatty acid compositions of TAGs produced from acetate and toluene were different. At the end of cultivation, the mole fraction of C17:0, one of odd-carbon number fatty acids, was 5% on toluene or 10% on acetate while the mole fraction of 3HV in PHBV from toluene was as high as that in PHBV from acetate, suggesting that a C5 intermediate of toluene degradation might directly become a precursor of 3HV whereas propionyl-CoA is required for the incorporation of C17:0 into TAGs.     

Phenolic composition,antioxidant and acetylcholinesterase inhibitory activities of Sclerocarya birrea and Harpephyllum caffrum (Anacardiaceae) extracts

Total phenolic content, proanthocyanidins, gallotannins, flavonoids, and antioxidant activities of Sclerocarya birrea and Harpephyllum caffrum methanolic extracts were evaluated using in vitro assays. S. birrea young stem extract contained the highest levels of total phenolic content (14.15 ± 0.03 mg GAE/g), flavonoids (1.21 ± 0.01 mg CE/g) and gallotannins (0.24 ± 0.00 mg GAE/g). H. caffrum stem bark extract had the highest content of proanthocyanidins (1.47%). The EC₅₀ values of the extracts in the DPPH free radical scavenging assay ranged from 4.26 to 6.92 μg/ml, compared to 6.86 μg/ml for ascorbic acid. A dose-dependent linear curve was obtained for all extracts in the ferric-reducing power assay. Dichloromethane and methanol extracts exhibited dose-dependent acetylcholinesterase inhibitory activity. Similarly, all extracts exhibited high antioxidant activity comparable to butylated hydroxytoluene based on the rate of β-carotene bleaching (84.1–93.9%). The two Anacardiaceae species provide a source of natural antioxidants and acetylcholinesterase inhibitors, and may be beneficial to the health of consumers.     

The formation of 2,5-dimethyl-4-hydroxy-2H-furan-3-one by cell-free extracts of Methylobacterium extorquens and strawberry (Fragaria × ananassa cv. Elsanta)

The formation of 2,5-dimethyl-4-hydroxy-2H-furan-3-one (DMHF), using the bacterial cells of Methylobacterium extorquens (strain with CABI registration number: IMI 369321) and strawberry cells (Fragaria × ananassacv. Elsanta), was studied. A combined mixture of the bacterial cell-free extract plus the strawberry extract was used as the enzymatic source. The pH of all the experimental aliquots was pH 6.0. The studied substrates were short-chain alcohols, carbohydrates and one amino acid. DMHF was analyzed by HPLC with UV detection at 280 nm. When 1,2-propanediol, 1-propanol, sucrose and fructose were used as substrates, the best production rates of DMHF were achieved, having a combined mixture of the bacterium and strawberry (minus the achenes) as the enzymic source. On the other hand, glucose, 1,2-propanediol, 2-propanol and fructose were the best substrates for the production of DMHF when a combined mixture of the bacterium and achenes served as the enzymic source. All results were obtained by comparing the production of DMHF from the strawberry extract alone and the bacterium plus the strawberry, without the achenes (in the first case) and the production of DMHF from the achene extract alone and the bacterium plus the achenes (in the second case). Moreover, the dehydrogenation activities of these three enzyme sources were determined by measuring the absorption of NADH at 340 nm. The substrate which was used for this series of the experiments was 1,2-propanediol. These results indicate the undoubted cooperation between the plant and the bacterial cells. This paper reports, for the first time, the formation of DMHF by bacterial and plant cell-free extracts.     

A novel thermostable chitinase (PJC) from pomegranate (Punica granatum) juice

A novel chitinase was isolated and purified to its homogeneity from pomegranate juice by a combination of ammonium sulphate precipitation and ion-exchange chromatography. The pomegranate juice chitinase (PJC) was purified to specific activity of 14.5 U/mg and a recovery of 34%. The monomeric protein migrated on SDS–PAGE at 29 kDa. The enzyme was found to be glycosylated (7.2%). It exhibited optimal activity at pH 4.5 and 70 °C. The enzyme was stable in the pH range 3.0–9.0 and up to 65 °C. The internal peptide sequence results suggest that the purified PJC shared high homology with class III chitinases of other known plant chitinases. The purified enzyme could hydrolyse colloidal chitin to its oligomers. It did not exhibit any antifungal activity.     

Determination of stilbenes in Sicilian pistachio by high-performance liquid chromatographic diode array (HPLC-DAD/FLD) and evaluation of eventually mycotoxin contamination

In this work, we have investigated about the presence of several natural stilbenes in 12 samples of pistachios harvested from 10 different farms of Sicily (Bronte and Agrigento). At the same time, we have evaluated the relation between the stilbenes synthesis and the possible contamination of mycotoxin produced by Aspergillus flavus and Aspergillus parasiticus. We have found two types of stilbenes in the samples of pistachios examined: trans-resveratrol and trans-resveratrol-3-O-β-glucoside (trans-piceid). Their concentration ranged from 0.07 to 0.18 mg/kg (av. = 0.12 ± 0.03 mg/kg) for trans-resveratrol, from 6.20 to 8.15 mg/kg (av. = 6.97 ± 0.55 mg/kg) for trans-piceid and from 6.38 to 8.27 mg/kg (av. = 7.09 ± 0.54 mg/kg) for total resveratrol.     

Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu (Morinda citrifolia)

An investigation into the effects of ethanol concentration (0–100%, v/v), extraction time (20–120 min) and extraction temperature (25–65 °C) on the extraction of phenolic antioxidants from mengkudu (Morinda citrifolia) was performed using a single-factor experiment. Total phenolic content (TPC) and total flavonoid content (TFC) assays were used for determination of phenolic compounds. Antioxidant capacity was evaluated by measuring the scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals. Experimental results showed that extraction conditions had significant effect on extraction of phenolic compounds and antioxidant capacities. The optimised conditions were 40% ethanol for 80 min at 65 °C, with values of 919.95 mg GAE/100 g DW for TPC, 472.73 mg CE/100 g DW for TFC, 791.71 μmol TEAC/100 g DW for ABTS and 1928.5 μmol TEAC/100 g DW for DPPH. TPC was significantly correlated with DPPH under the effects of ethanol concentration (r = 0.932) and extraction time (r = −0.938).     

Precise control of repeating unit composition in biodegradable poly(3-hydroxyalkanoate) polymers synthesized by Escherichia coli

The composition of medium-chain-length (MCL) poly(3-hydroxyalkanoate) (PHA) biopolymers is normally an uncontrollable random mixture of repeating units with differing side chain lengths. Attempts to generate MCL PHA homopolymers and control repeating unit composition have been published in native PHA-producing organisms but have limited ranges for the different sizes of repeating units that can be synthesized. In this study, a new Escherichia coli-based system that exhibits control over repeating unit composition for both MCL PHAs and short-chain-length (SCL) PHAs has been developed, covering an unprecedented range of repeating units. The fadB and fadJ genes from the β-oxidation pathway were eliminated from the chromosome of E. coli LS5218. The subsequent blockage in β-oxidation caused a buildup of enoyl-CoA intermediates, which were converted to PHAs by an (R)-specific enoyl-CoA hydratase (PhaJ4) and PHA synthase [PhaC1(STQK)] expressed from a plasmid DNA construct. Fatty acid substrates were converted to PHAs with repeating units equal in the number of carbon atoms to the fatty acid substrate. The broad substrate specificities of the PhaJ4 and PhaC1(STQK) enzymes allowed for the production of homopolymers with strict control over the repeating unit composition from substrates of four to twelve carbons in length. Polymers were purified and analyzed by GC, GC-MS, and NMR for structural composition and by DSC, TGA, and GPC for thermal and physical characteristics. This study marks the development of the first single biological system to achieve consistent repeating unit control over such a broad range of repeating units in PHAs.     

Matrix metalloproteinases (MMPs) inhibitory effects of an octameric oligopeptide isolated from abalone Haliotis discus hannai

Abalone (Haliotis discus hannai) is a marine gastropod, and an important fishery and food industrial resource that is massively maricultured in Asia, Africa, Australia and America. However, its health benefits have rarely been studied for nutraceutical and pharmaceutical application. In this study, the purified abalone oligopeptide (AOP) with anti-matrix metalloproteinases (anti-MMPs) effects was isolated from the digests of abalone intestine using recycle HPLC with a JAI W253 column and an OHpak SB-803 HQ column. The AOP was identified as Ala-Glu-Leu-Pro-Ser-Leu-Pro-Gly (MW = 782.4 Da) with a de novo peptide sequencing technique using a tandem mass spectrometer. The AOP exhibited a specific inhibitory effect against MMP-2/-9 activity and attenuated protein expression of p50 and p65 in the human fibrosarcoma (HT1080) cells, dose-dependently. The results presented illustrate that the AOP could inhibit MMP-2/-9 expression in HT1080 cells via the nuclear factor-kappaB (NF-κB)-mediated pathway. This data suggest that the AOP from H. discus hannai intestine may possess therapeutic and preventive potential for the treatment of MMPs-related disorders such as angiogenesis and cardiovascular diseases.     

Cloning and Sequence Analysis of the estA gene encoding enzyme for producing (R)-β-acetylmercaptoisobutyric acid from Pseudomonas aeruginosa 1001

The estA gene encoding the enzyme that catalyzes the production of (R)-β-acetylmercaptoisobutyric acid from (R,S)-ester from Pseudomonas aeruginosa 1001, was cloned in Escherichia coli and its nucleotide sequence was determined, revealing the presumed open reading frame encoding a polypeptide of 316 amino acid residues (948 nucleotides). The overall A+T and C+G compositions were 32.59% and 67.41%, respectively. The amino acid sequence of the estA gene product showed a significant similarity with that of the triacylglycerol lipase from Psychrobacter immobilis (38% identity), triacylglycerol lipase from Moraxella sp. (36% identity), and two forms of carboxyl esterases from Acinetobacter calcoaceticus (17% and 17% identities). The deduced amino acid sequences have a pentapeptide consensus sequence, G-X-S-X-G, having an active serine residue, and another active site, dipeptides H-G, located at 70–100 amino acids upstream of the G-X-S-X-G consensus sequence.     

Characterisation of 2-Cys peroxiredoxin isozyme (Prx1) from Taiwanofungus camphorata (Niu-chang-chih): Expression and enzyme properties

Peroxiredoxins (Prxs) are a family of antioxidant peroxidases. The functions of Prxs comprises of cell protection against oxidative stress and regulation of cell proliferation. A putative 2-Cys Prx isozyme (Prx1) cDNA was cloned from Taiwanofungus camphorata (commonly known as Niu-chang-chih in Taiwan). The deduced amino acid sequence is conserved amongst the reported Prxs. A 3-D homology structure was created for this Prx1. To characterise the T. camphorata Prx1, the coding region was subcloned into a pAVD10 and transformed into Escherichia coli. The recombinant 6His-tagged Prx1 was expressed and purified by Ni²⁺-nitrilotriacetic acid sepharose. The purified enzyme showed two forms using a 15% SDS–PAGE. The enzyme retained 60% activity at 60 °C for 2.5 min. The enzyme was stable under a broad pH range from 5 to 11. The enzyme showed 57% activity after 40 min of incubation at 37 °C with trypsin. The ability of the enzyme to protect intact supercoiled plasmid DNA from ^·OH induced nicking was demonstrated.     

Characterisation of an acidic peroxidase from papaya (Carica papaya L. cv Tainung No. 2) latex and its application in the determination of micromolar hydrogen peroxide in milk

An acidic peroxidase isoform, POD-A, with a molecular mass of 69.4 kDa and an isoelectric point of 3.5 was purified from papaya latex. Using o-phenylenediamine (OPD) as a hydrogen donor (citrate–phosphate as pH buffer), the optimum pH for the function of POD-A was 4.6, and the optimum temperature was 50 °C. The peroxidase activity of POD-A toward hydrogen donors was both pH- and concentration-dependent. Under optimal conditions, POD-A catalysed the oxidation of OPD at higher rates than pyrogallol, catechol, quercetin and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). The chemical modification reagents N-bromosuccinimide and sodium azide significantly inhibited POD-A activity. The results of kinetic studies indicated that POD-A followed a ping-pong mechanism and had a K_m value of 2.8 mM for OPD. Using CPC silica-immobilised POD-A for the determination of micromolar H₂O₂ in milk, the lower limit of determination was 0.1 μM, and the recoveries of added H₂O₂ were 96–109%.     

Chemical composition,fatty acid content and antioxidant potential of meat from goats supplemented with Moringa (Moringa oleifera) leaves,sunflower cake and grass hay

The present study determined the chemical composition, fatty acid (FA) content and antioxidant capacity of meat from goats supplemented with Moringa oleifera leaves (MOL) or sunflower cake (SC) or grass hay (GH). The meat from goat supplemented with MOL had higher concentrations of total phenolic content (10.62 ± 0.27 mg tannic acid equivalent E/g). The MOL significantly scavenged 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic-acid (ABTS) radical to 93.51 ± 0.19% (93.51 ± 0.19%) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical to 58.95 ± 0.3% than other supplements. The antioxidative effect of MOL supplemented meat on catalase (CAT), reduced glutathione (GSH), superoxide dismutase (SOD) and lipid oxidation (LO) was significantly (P < 0.05) higher than other meat from goat feed on grass hay or those supplemented with sunflower seed cake. The present study indicated that the anti-oxidative potential of MOL may play a role in improving meat quality (chemical composition, colour and lipid stability).     

Açaí (Euterpe oleraceae) ‘BRS Pará’: A tropical fruit source of antioxidant dietary fiber and high antioxidant capacity oil

This article reports a study of the concentrations of dietary fiber (DF) and antioxidant capacity in fruits (pulp and oil) of a new açaí (Euterpe oleraceae) cultivar—‘BRS-Pará’, with a view to determine the possibility of using it as a source of antioxidants in functional foods or dietary supplements. Results show that ‘BRS-Pará’ açaí fruits has a high content of DF (71% dry matter) and oil (20.82%) as well as a high antioxidant capacity in both defatted matter and oil. ‘BRS-Pará’ Açaí fruits can be considered as an excellent source of antioxidant dietary fiber. Antioxidant capacity of açaí ‘BRS-Pará’ oil by DPPH assay was higher (EC₅₀ = 646.3 g/g DPPH) than extra virgin olive oil (EC₅₀ = 2057.27 g/g DPPH). These features provide açaí ‘BRS-Pará’ fruits with considerable potential for nutritional and health applications.     

Purification and characterization of an antioxidant protein (∼16 kDa) from Terminalia chebula fruit

Terminalia chebula fruit is used as folk medicine in India and Southeast Asia. An antioxidant protein was isolated by bioassay guided fractionation of T.chebula fruit by homogenizing in the citrate phosphate buffer. The isolated protein (TCP-III) obtained from fruit was purified by gel chromatography and preparative HPLC, showed apparent molecular weight of 16 kDa by SDS-PAGE and MALDI-TOF/MS analyses. Amino acid sequence obtained by LC-MS^E analysis showed homology with the predicted protein fragments of Populus trichocarpa, putative uncharacterized protein fragments from Oryza sativa and with fragments of 17 kDa thylakoid lumenal protein from Spinacia oleracea. TCP-III exhibited significant radical scavenging in DPPH, NO, H₂O₂ and ABTS assays. In addition, TCP-III inhibited oxidation of linoleic acid in β-carotene bleaching assay, reduced ferric ions and chelated ferrous ions. The present finding demonstrates uniquely, for the first time, characterization of an antioxidant protein from T. chebula fruit.     

Isolation of thermophiles degrading poly( -lactic acid)

A thermophile, strain 93, which degrades poly( -lactic acid) (PLA) film was isolated from 144 soil samples obtained from different locations by cultivation using an enrichment culture medium at 60°C. Under this temperature condition, the strain grew on PLA and the dissolved total organic carbon (TOC) concentration in the medium changed according to the growth stage, i.e., after the TOC rapidly reached the minimum, it increased rapidly until it reached the peak and then decreased thereafter. For residual PLA, reduced viscosity decreased rapidly but weight decreased initially slowly with a gradually increasing rate. Gel permeation chromatograms showed a marked decrease in the main peak and the appearance of a new peak in the low molecular weight region. The strain was identified as Bacillus brevis, which has an optimum growth temperature of around 58°C.