Synthetic Biology and Metabolic Engineering for Marine Carotenoids: New Opportunities and Future Prospects

Carotenoids are a class of diverse pigments with important biological roles such as light capture and antioxidative activities. Many novel carotenoids have been isolated from marine organisms to date and have shown various utilizations as nutraceuticals and pharmaceuticals. In this review, we summarize the pathways and enzymes of carotenoid synthesis and discuss various modifications of marine carotenoids. The advances in metabolic engineering and synthetic biology for carotenoid production are also reviewed, in hopes that this review will promote the exploration of marine carotenoid for their utilizations.     

Genetic evidence of stable northward extension of Pinus thunbergii Parl. forests in the Democratic People’s Republic of Korea

Genetic Resources and Crop Evolution - Japanese black pine (Pinus thunbergii Parl.) with high salt tolerance may be an important constitutive element sustaining terrestrial ecosystem by playing a...     

Manipulation of vertical fence wake using passive vortex generators

The effect of streamwise vortices generated from passive vortex generators was investigated to manipulate the separation bubble behind the vertical fence. The experiments were carried out in a circulating water channel and the velocity fields were measured using 2D and stereoscopic PIV method.The distance between the vortex generator and fence and the effect of the Reynolds number were investigated. In addition,the effect of boundary layer thickness was also investigated. The averaged recirculation lengths were compared with that of uncontrolled fence flow. The results showed the oscillatory variation of recirculation region appeared under the existence of vortex generators. The reduction of the separation bubble became larger when the fence was submerged in the thick boundary layer with increasing the distance between the generator and fence. When the boundary layer is thin,vortex generator can only suppress the separation bubble under the specific condition.     

Long-term evaluation of extruded diets compared with raw fish moist diet for growing Korean rockfish, Sebastes schlegeli (Hilgendorf)

A long‐term experiment was conducted in sea cages to compare three experimental extruded pellet (EP) diets with a raw fish moist pellet (MP) diet for Korean rockfish, Sebastes schlegeli (Hilgendorf). White fishmeal (WFM), soybean meal (SM) and fishmeal analogue (BAIFA‐M?)‐based experimental EP diets were formulated to be isonitrogenous and isocaloric with 50% crude protein (CP) with available energy of 16.7 kJ g^?1 diet. An MP diet made of 80% frozen horse mackerel + 20% commercial binder meal was also included in the experiment. The proportions of the major protein ingredients in three experimental EP diets were as follows: WFM diet, 100% WFM; SM diet, 70% WFM + 30% SM; BAIFA‐M? diet, 70% WFM + 30% BAIFA‐M?. The experiment was divided into three periods according to fish size: period I, initial weight of 20.2 ± 3.6 g (mean ± SD); period II, initial weight of 57.6 ± 4.7 g (mean ± SD); period III, initial weight of 96.3 ± 6.9 g (mean ± SD). There were no significant differences in weight gain, feed efficiency ratio, thermal‐unit growth coefficient, specific growth rate, haemoglobin, or haematocrit among fish that were fed the four diets during all three periods. However, fish that were fed the MP diet had a lower survival rate than the fish that were fed the other three EP diets for all periods. These results strongly suggest that EP diets could be developed to replace MP diet for the grow‐out stage of production of Korean rockfish without adverse effects on growth performance.     

Allelic losses and gains during translocations of a high conservation value fish,Coregonus lavaretus

1. The use of translocations to establish new or ‘refuge’ populations for species with high conservation value is controversial but widely used in conservation management. One of the risks of this approach is that an establishing population does not adequately capture the genetic diversity of the donor gene pool. This effect, rarely examined, is tested here.
2. In this study the genetic consequences of two conservation translocations after five generations (16 years) of the European whitefish, Coregonus lavaretus, were quantified. Both translocations were made using almost the same genetic groups and thus represent a partly replicated natural study.
3. Analysis of 12 informative microsatellites showed that expected heterozygosity, the mean number of alleles per locus and allelic richness did not differ between donor and translocated populations. There was also no loss of heterozygosity in the translocated populations, nor deviations from Hardy–Weinberg equilibrium expectations, nor signs of linkage disequilibrium.
4. All populations were genetically differentiated but pairwise F_ST values were low, indicating that the magnitude of divergence was small.
5. There was no evidence of inbreeding but there were significant differences in private allelic richness between donor and translocated populations. Of 50 alleles found in the donor population, 16% of the rarer alleles were lost in one translocated population and 8% in the other.
6. Allele loss without a reduction in heterozygosity strongly points to stochastic drift effects having occurred following translocation. The evidence indicates that alleles that were not detected in the donor population have arisen de novo in the translocated populations.
7. It is concluded that conservation translocations comprising even a modest number of propagules can successfully capture a high proportion of genetic variation of the host population, and that reduced genetic variation in the translocated population may be mitigated by the emergence of new variation over short time periods.

     

Prevalence and different characteristics of two serotypes of Streptococcus parauberis isolated from the farmed olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in Korea

The prevalence of two serotypes of Streptococcus parauberis isolated from the olive flounder, Paralichthys olivaceus, was evaluated in a total of 29 isolates between 2003 and 2010 in Korea. Streptococcus parauberis isolates were divided into two serologically distinct types (serotype 1 and serotype 2), except for one strain (S1091), using an agglutination assay with rabbit antiserum, and serotype 1 was identified as the dominant type (24 of 29 isolates) in this study. To identify the characteristics of the two serotypes of S. parauberis, we conducted a biochemical test using the API 20 Strep kit, a transmission electron microscopy (TEM) assay, sequence analysis of 16S‐23S rRNA intergenic spacer region (ISR) and a pathogenicity test. In TEM, both serotypes possessed polysaccharide capsule layers around the cell surface when bacterial cells were treated with a homologous serotype of rabbit antiserum. However, we were unable to discriminate serotype‐specific biochemical characteristics and genetic characteristics of 16S‐23S rRNA ISR between the two serotypes. In the pathogenicity test, the serotype 1 strains induced significantly higher mortality than the serotype 2 strains in olive flounder when experimentally inoculated via the intraperitoneal route.     

Effect of taurine supplemented practical diet on growth performance and taurine contents in whole body and tissues of juvenile Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

ABSTRACT:   The effect of dietary taurine on juvenile Japanese flounder was determined by feeding three taurine-supplemented experimental diets (TAU) and a commercial diet (CD) to evaluate a practical diet for juvenile Japanese flounder. Juvenile Japanese flounder were reared on the three experimental diets supplemented with taurine at 0, 0.5, 1.0% and CD. These diets were fed to juvenile Japanese flounder of an initial mean body weight of 0.2 g for 6 weeks at 20°C and the taurine contents of the whole body and tissues were analyzed. The final average body weight of juvenile Japanese flounder fed the 1.0% TAU was significantly higher than that of the other groups. Taurine contents in the whole body and tissues increased with the increase in dietary taurine level. These results indicate that juvenile Japanese flounder require at least 15 mg/g taurine in the diet, even though a combined mix of fish, krill and squid meal was the main protein source in the experimental diets.     

Chemical profiling with modeling differentiates wild and farm-raised salmon

Classifications of fish production methods, wild or farm-raised salmon, by elemental profiles or C and N stable isotope ratios combined with various modeling approaches were determined. Elemental analysis (As, Ba, Be, Ca, Co, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, Sr, Ti, and Zn) of wild and farm-raised salmon samples was performed using an inductively coupled plasma atomic emission spectroscopy. Isotopic and compositional analyses of carbon and nitrogen were performed using mass spectrometry as an alternative fingerprinting technique. Each salmon (king salmon, Oncorhynchus tshawytscha ; coho salmon, Oncorhynchus kisutch ; Atlantic salmon, Salmo salar ) was analyzed from two food production practices, wild and farm raised. Principal component analysis (PCA) and canonical discriminant analysis (CDA) were used for data exploration and visualization. Five classification modeling approaches were investigated: linear discriminate function, quadratic discriminant function, neural network, probabilistic neural network, and neural network bagging. Methods for evaluating model reliability included four strategies: resubstitution, cross-validation, and two very different test set scenarios. Generally speaking, the models performed well, with the percentage of samples classified correctly depending on the particular choice of model and evaluation method used.     

Antioxidant activity of glyceollins derived from soybean elicited with Aspergillus sojae

The extract of soybean exposed to biotic elicitors such as food-grade fungus is known to have antioxidant activity. Glyceollins were major bioactive compounds present in soybean elicited by fungi and shown to have antifungal and anticancer activities. The purpose of present study was to evaluate the antioxidant activities of glyceollins by measuring ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, singlet oxygen quenching, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, hydroxyl radical scavenging activity, and lipid peroxidation inhibition. In addition, the antioxidant potential of glyceollins were measured by a fluorescent probe, 2',7'-dichlorofluorescin diacetate (DCFDA), and dihydroethidium (DHE) in mouse hepatoma hepa1c1c7 cells in which they were insulted with H2O2 to generate reactive oxygen species (ROS). Glyceollins showed a strong reducing power and inhibited lipid peroxidation, with significant scavenging activities of radicals including singlet oxygen, superoxide anion, ABTS, and DPPH. We also found that glyceollins significantly suppressed H2O2-induced ROS production in hepa1c1c7 cells. Therefore, glyceollins deserve further study as natural antioxidants and nutraceuticals.     

Attenuation of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced gap junctional intercellular communication (GJIC) inhibition in MCF-10A cells by c9,t11-conjugated linoleic acid

The protective effect of c9,t11-conjugated linoleic acid (CLA) on the inhibition of gap junctional intercellular communication (GJIC) was examined in a human mammary epithelial cell line (MCF-10A) treated with 12-O-tetradecanoylphorbol-13-acetate (TPA), relative to t10,c12-CLA isomer. TPA inhibited GJIC in a dose-dependent and reversible manner and was associated with connexin 43 phosphorylation. Pretreatment of 20 μM c9,t11-CLA for 24 h prior to 60 nM TPA for 1 h prevented the inhibition of GJIC by reducing the phosphorylation of connexin 43 via suppressing extracellular signal-regulated kinases (ERK1/2) activation. Reactive oxygen species (ROS) accumulation by TPA was attenuated by c9,t11-CLA. The efficacy of c9,t11-CLA in protecting inhibition of GJIC, connexin 43 phosphorylation, and ROS production was superior to that of t10,c12-CLA. These results suggest that c9,t11-CLA, including t10,c12-CLA, prevents the carcinogenesis of MCF-10A cells by protecting down-regulation of GJIC during the cancer promotion stage, and lack of their toxicities could be an excellent indicator for the chemoprevention of breast cancer.