Denovo production of resveratrol by engineered Saccharomyces cerevisiae W303-1a using pretreated Gracilaria corticata extracts.

OBJECTIVE: Assembly and construction of resveratrol production pathway in Saccharomyces cerevisiae for denovo production of resveratrol using seaweed extract as fermentation medium. RESULTS: Genes involved in the production of resveratrol from tyrosine pathway, tyrosine ammonia lyase (FTAL) gene from Flavobacterium johnsoniae (FjTAL), the 4-coumarate:CoA ligase gene from Arabidopsis thaliana (4CL1) and the stilbene synthase gene from Vitis vinifera (VvSTS) were introduced into low copy, high copy and integrative vector and transformed into S. cerevisiae W303-1a. The resulting strains W303-1a/pARS-res5, W303-1a/2µ-res1 and W303-1a/IntUra-res9 produced a level of 2.39 ± 0.01, 3.33 ± 0.03 and 8.34 ± 0.03 mg resveratrol l-1 respectively. CRISPR mediated integration at the δ locus resulted in 17.13 ± 1.1 mg resveratrol l-1. Gracilaria corticata extract was tested as a substrate for the growth of transformant to produce resveratrol. The strain produced a comparable level, 13.6 ± 0.54 mg resveratrol l-1 when grown in seaweed extract medium. CONCLUSIONS: The strain W303-1a/IntδC-res1 utilized Gracillaria hydrolysate and produced 13.6 ± 0.54 mg resveratrol l-1 and further investigations are being carried out focusing on pathway engineering and optimization of process parameters to enhance resveratrol yield.

In vitro fermentation of Gracilaria lemaneiformis and its sulfated polysaccharides by rabbitfish gut microbes.

This study intended to characterize the Gracilaria lemaneiformis (SW)-derived polysaccharide (GLP) and explore the fermentation aspects of SW and GLP by rabbitfish (Siganus canaliculatus) intestinal microbes. The GLP was mainly composed of galactose and anhydrogalactose (at 2.0:0.75 molar ratio) with the linear mainstay of α-(1 â†’ 4) linked 3,6-anhydro-α-l-galactopyranose and ß-(1 â†’ 3)-linked galactopyranose units. The in vitro fermentation results showed that the SW and GLP could reinforce the short-chain fatty (SCFAs) production and change the diversity and composition of gut microbiota. Moreover, GLP boosted the Fusobacteria and reduced the Firmicutes abundance, while SW increased the Proteobacteria abundance. Furthermore, the adequacy of feasibly harmful bacteria (such as Vibrio) declined. Interestingly, most metabolic processes were correlated with the GLP and SW groups than the control and galactooligosaccharide (GOS)-treated groups. In addition, the intestinal microbes degrade the GLP with 88.21 % of the molecular weight reduction from 1.36 × 105 g/mol (at 0 h) to 1.6 × 104 g/mol (at 24 h). Therefore, the findings suggest that the SW and GLP have prebiotic potential and could be applied as functional feed additives in aquaculture.

Digestion characteristics of polysaccharides from Gracilaria lemaneiformis and its interaction with the human gut microbiota.

The health effects of polysaccharides have attracted lots of attention, but the exact mechanism remains unclear. This study indicated that polysaccharides from Gracilaria lemaneiformis (GLPs) tolerated the conditions of mouth, stomach, and small intestine, and it reached the colon integrally, where it increased the production of short chain fatty acids, altered the gut microbiota, and especially increased the level of Bacteroides. To explore the underlying mechanism, hundreds of Bacteroides strains were isolated from the human feces and identified by MALDI-TOF/MS. It showed that Bacteroides species profile was different between individuals, revealing an inherent difference in the human gut microbiota. The use of Bacteroides on GLPs was species-dependent, and various small molecular GLPs fragments can be liberated from growth of Bacteroides species. On the other hand, Bacteroides species that unable to grow with GLPs can live in GLPs-derived fragments, forming a GLPs utilization network. It should be noted that small molecular GLPs fragments can be easier to be metabolized by intestinal microbes and have better effect on cellular response. It suggested that the effect of polysaccharides cannot only be attributed to modulation of the gut microbiota, but also associated with the effect of microbial degradation on GLPs own activities.

Effects of seawater sulfur starvation and enrichment on Gracilaria gracilis growth and biochemical composition.

The genus Gracilaria, largest biomass producer in coastal regions, encompasses a wide range of species including Gracilaria gracilis. Nowadays, there is a spate of interest in its culture in lagoon where the water sulfate concentration is variable. A laboratory culture was carried out to determine the sulfate concentration effect on their growth as well as their biochemical composition, which were 2.5, 27 or 50 mM, referred to as SSS (sulfur starved seawater), SW (seawater) and SES (sulfur enriched seawater).We found that the sulfate content of the surrounding medium is a key parameter influencing both the alga growth and its composition. However, seawater proved to be the most suitable environment to sustain alga growth, proteins, R-phycoerythrin and agar yields, but sulfur enrichment and starvation affects them. The sulfate degree of agar and therefore its quality is related to the medium sulfate concentration. We conclude that sulfur starvation (2.5 mM) for three weeks, led to severe growth retardation, lower agar yield and quality and indicated the limit potential of G. gracilis for mariculture under these conditions. These results demonstrated that the success of G. gracilis culture in the lagoon is feasible if sulfate concentration is closer to that of seawater.

Single-step purified R-phycoerythrin transmits cellular imaging functionalities in vitro.

A single-step and rapid chromatographic method-based purification of Gracilaria corticata (J. Agardh) R-phycoerythrin (R-PE) was attained using polyacrylamide gel electrophoresis (PAGE) technique without affecting structural integrity. The purified R-PE had a characteristic UV-Vis spectrum with three absorbance maxima at 496, 535, and 565 nm, and fluorescence at 575 nm. R-PE was obtained with a purity index of 4.2 and a recovery yield of 44.3%. SDS-PAGE analysis exhibited three sub-units i.e., 18, 21, and 31 kDa, which corresponds to α, ß, and γ, respectively. This report's purification process was considered less time-consuming and could be efficiently applied to purify phycobiliproteins. The purified R-PE showed optimal stability up to 6 h at pH 7.0 when exposed to light (3000 lx), while the temperature at which the maximum stability was retained was at 20 °C. The cellular imaging property of R-PE was effectively implemented to evaluate its credentials without affecting the cell proliferation of Vero and Hep-2 cell lines with the higher IC50 concentrations in vitro. Under fluorescence microscopy and flow cytometry analysis, purified R-PE displayed the characteristic affinity towards cell imaging functions in preliminary in vitro studies.

Synthesis, characterization, in vitro antioxidant and hypoglycemic activities of selenium nanoparticles decorated with polysaccharides of Gracilaria lemaneiformis.

Under a simple redox system of selenite and ascorbic acid, we used Gracilaria lemaneiformis polysaccharides (GLPs) as a stabilizer and dispersing agent to generate well-dispersed and stable selenium nanoparticles (SeNPs). The size, stability, morphology and physicochemical properties of GLPs-SeNPs were characterized by dynamic light scattering (DLS), scanning electron microscopy (SEM), transmission electron microscopy (TEM), Ultraviolet-visible spectroscopy (UV-vis), Fourier transform infrared (FT-IR) spectra, energy dispersive X-ray (EDX), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Thermogravimetric (TG). The results showed that orange-red, amorphous, zero-valent and spherical GLPs-SeNPs with mean diameter of approximately 92.5 nm were successfully prepared, which exhibited good storage stability at 4 °C and remaining highly stable at different ion strengths and pH. The 2,2-diphenyl-1-pycrylhydrazyl (DPPH), 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) and superoxide anion radical (O2•-) radical scavenging ability of GLPs-SeNPs were higher than those of bare SeNPs, GLPs and sodium selenite (Na2SeO3), and could reach 103.41%, 94.23%, 86% at a concentration of 1.5 mg/mL, respectively. Besides, GLPs-SeNPs also showed higher inhibitory effects on α-amylase and α-glucosidase. In vitro cytotoxicity assay and hemolysis activity examinations indicated that GLPs-SeNPs have excellent biocompatibility. Therefore, the GLPs-SeNPs might be used as a potential antioxidant agent and antidiabetic agent for food and medical applications.

On the Health Benefits vs. Risks of Seaweeds and Their Constituents: The Curious Case of the Polymer Paradigm.

To exploit the nutraceutical and biomedical potential of selected seaweed-derived polymers in an economically viable way, it is necessary to analyze and understand their quality and yield fluctuations throughout the seasons. In this study, the seasonal polysaccharide yield and respective quality were evaluated in three selected seaweeds, namely the agarophyte Gracilaria gracilis, the carrageenophyte Calliblepharis jubata (both red seaweeds) and the alginophyte Sargassum muticum (brown seaweed). It was found that the agar synthesis of G. gracilis did not significantly differ with the seasons (27.04% seaweed dry weight (DW)). In contrast, the carrageenan content in C. jubata varied seasonally, being synthesized in higher concentrations during the summer (18.73% DW). Meanwhile, the alginate synthesis of S. muticum exhibited a higher concentration (36.88% DW) during the winter. Therefore, there is a need to assess the threshold at which seaweed-derived polymers may have positive effects or negative impacts on human nutrition. Furthermore, this study highlights the three polymers, along with their known thresholds, at which they can have positive and/or negative health impacts. Such knowledge is key to recognizing the paradigm governing their successful deployment and related beneficial applications in humans.

The Use of Invasive Algae Species as a Source of Secondary Metabolites and Biological Activities: Spain as Case-Study.

In the recent decades, algae have proven to be a source of different bioactive compounds with biological activities, which has increased the potential application of these organisms in food, cosmetic, pharmaceutical, animal feed, and other industrial sectors. On the other hand, there is a growing interest in developing effective strategies for control and/or eradication of invasive algae since they have a negative impact on marine ecosystems and in the economy of the affected zones. However, the application of control measures is usually time and resource-consuming and not profitable. Considering this context, the valorization of invasive algae species as a source of bioactive compounds for industrial applications could be a suitable strategy to reduce their population, obtaining both environmental and economic benefits. To carry out this practice, it is necessary to evaluate the chemical and the nutritional composition of the algae as well as the most efficient methods of extracting the compounds of interest. In the case of northwest Spain, five algae species are considered invasive: Asparagopsis armata, Codium fragile, Gracilaria vermiculophylla, Sargassum muticum, and Grateulopia turuturu. This review presents a brief description of their main bioactive compounds, biological activities, and extraction systems employed for their recovery. In addition, evidence of their beneficial properties and the possibility of use them as supplement in diets of aquaculture animals was collected to illustrate one of their possible applications.

Stability, kinetics, and application study of phycobiliprotein pigments extracted from red algae Gracilaria gracilis.

Phycobiliprotein (PBP) pigments were extracted from red algae Gracilaria gracilis through maceration in phosphate buffer using previously optimized conditions. The stability of PBPs in the extracts was assessed by monitoring the extracts at different pHs and temperatures for 10 days. Since phycoerythrin (PE) is the main PBP present in G. gracilis, PE content was spectroscopically determined and used as a response factor. Kinetic modeling was used to describe PE degradation under different ranges of T and pH. The pigment extracts presented higher stability at pH 6.9 and -20 °C. PE was semipurified by precipitation with ammonium sulphate 65% followed by dialysis against water until a purity index of 0.7. The pigment was successfully applied as colorant in pancakes and yogurts with a pigment concentration of 0.15%. This study highlights the potential of PE pigments extracted from G. gracilis for applications in food products. PRACTICAL APPLICATION: Phycobiliprotein pigments were extracted from red algae Gracilaria gracilis through maceration in phosphate buffer. The stability of the pigment was evaluated at different pHs and temperatures, presenting higher stability at neutral pH and low temperatures. The pigment was successfully applied as colorant in pancakes and yogurts with a low pigment concentration. This study highlights the potential of phycobiliprotein pigments extracted from G. gracilis for applications in food products.

Effect of Lagoon and Sea Water Depth on Gracilaria gracilis Growth and Biochemical Composition in the Northeast of Tunisia.

This study evaluated the growth and biochemical composition of farming Gracilaria gracilis (Stackhouse) M. Steentoft, L. M. Irvine & W. F. Farnham in the Bizerte Lagoon (BL) and Bizerte Bay (BB) in the North Coast of Tunisia, using lantern nets. Effects of site and depth on alga daily growth rate (DGR) and biochemical composition were investigated. The DGR was affected by culture site (1.42 ± 0.65% day-1 and 1.19 ± 0.34% day-1 for the BL and the BB respectively). Agar yield, was higher (p < 0.05) in the BB than the BL (23.31 ± 2.64% vs. 19.19 ± 2.32%) with a higher (p < 0.05) 3,6-anhydrogalactose (3,6-AG) contents (41.37 ± 3.68% vs 23.30 ± 5.40%) and a lower (p < 0.05) sulphate degree (6 ± 2.00% vs 8.80 ± 0.86%). The proteins contents were independent of the site and depth of culture (20.74 ± 7.22% and 22.02 ± 6.34% for the BL and the BB respectively). R-phycoerythrin (R-PE) contents were significantly higher (p < 0.05) in the BB (0.86 ± 0.31 mg g-1) than those obtained in the BL (0.33 ± 0.12 mg g-1). The salinity, transparency, nitrate and ammonium were monitored in both sites, and their influences were discussed. Our results suggest that G. gracilis cultured in Bizerte Bay can be used in a cascading biorefinery approach.

Comparison of Ethanol Yield Coefficients Using Saccharomyces cerevisiae, Candida lusitaniae, and Kluyveromyces marxianus Adapted to High Concentrations of Galactose with Gracilaria verrucosa as Substrate.

The red seaweed Gracilaria verrucosa has been used for the production of bioethanol. Pretreatment for monosaccharide production was carried out with 12% (w/v) G. verrucosa slurry and 500 mM HNO3 at 121°C for 90 min. Enzymatic hydrolysis was performed with a mixture of commercial enzymes (Cellic C-Tec 2 and Celluclast 1.5 L; 16 U/ml) at 50°C and 150 rpm for 48 h. G. verrucosa was composed of 66.9% carbohydrates. In this study, 61.0 g/L monosaccharides were obtained from 120.0 g dw/l G. verrucosa. The fermentation inhibitors such as hydroxymethylfurfural (HMF), levulinic acid, and formic acid were produced during pretreatment. Activated carbon was used to remove HMF. Wildtype and adaptively evolved Saccharomyces cerevisiae, Candida lusitaniae, and Kluyveromyces marxianus were used for fermentation to evaluate ethanol production.

In vitro fermentation of Gracilaria lemaneiformis sulfated polysaccharides and its agaro-oligosaccharides by human fecal inocula and its impact on microbiota.

The fermentation behaviour of sulfated polysaccharides (GLP) and their agaro-oligosaccharides (GLO) derived from Gracilaria lemaneiformis were examined. During in vitro fermentation, GLP and GLO increased the concentrations of short chain fatty acids (SCFAs) and modulated the composition and diversity of gut microorganisms compared with control groups. GLP increased the abundance of Bacteroidetes and decreased the abundance of Firmicutes, while GLO increased the abundance of Firmicutes and Actinobacteria. Moreover, the abundances of potential pathogenic bacteria were reduced. Molecular weight and intrinsic viscosity of GLP decreased significantly from 2.15 × 105 to 1.22 × 105 Da, 374.45-113.91 mL/g, respectively. Furthermore, GLP was degraded into smaller degree of polymerization of oligosaccharides, with no significant change observed in GLO. Overall, this study revealed GLP and GLO could be beneficial for gastrointestinal tract by producing SCFAs and modulating intestinal microbes, indicating GLP and GLO are potentially sources of prebiotics in functional foods.

Marine Algae of the Genus Gracilaria as Multi Products Source for Different Biotechnological and Medical Applications.

BACKGROUND: Gracilaria has been shown to be an important source of marine bioactive natural biomaterials and compounds. Although there are no enough patents used Gracilaria worldwide, the current study tries to put the Gracilaria on the spot for further important patents in the future. OBJECTIVE: The current study investigates the pharmaceuticals and biochemical activity of Gracilaria because no previous studies have been carried out to examine the biochemical and pharmaceutical activates of Gracilaria from the Suez Canal of Egypt as an excellent source for bioactive compounds. METHODS: Different advanced experimental models and analytical techniques, such as cytotoxicity, total antioxidant capacity, anticancer, and anti-inflammatory profiling were applied. The phytochemical analysis of different constituents was also carried out. RESULTS: The mineral analysis revealed the presence of copper (188.3 ppm) and iron (10.07 ppm) in addition to a remarkable wealth of selenium and sulfur contents giving up to 36% of its dry mass. The elemental analysis showed high contents of sulfur and nitrogen compounds. The GCMS profiling showed varieties of different bioactive compounds, such as fatty acids, different types of carotenoids in addition to pigments, alkaloids, steroids. Many other compounds, such as carbohydrates and amino acids having antioxidant, anti-inflammatory, and antiviral activities, etc. were identified. The cytotoxicity activity of Gracilaria marine extract was very effective against cancerous cell lines and showed high ability as a potent antitumor due to their bioactive constituents. Specialized screening assays using two anticancer experimental models, i.e., PTK and SKH1 revealed 77.88% and 84.50% inhibition anticancer activity; respectively. The anti-inflammatory activities investigated using four different experimental models, i.e., COX1, COX2, IL6, and TNF resulted in 68%, 81.76%, 56.02% and 78.43% inhibition; respectively. Moreover, Gracilaria extracts showed potent anti-Alzheimer with all concentrations. CONCLUSION: Gracilaria proved to be a multi-product source of marine natural products for different biotechnological applications. Our recommendation is to investigate the Gracilaria bioactive secondary metabolites in order to create and innovate in more patents from current important seaweeds (Gracilaria).

Potential bioremediation effects of seaweed Gracilaria lemaneiformis on heavy metals in coastal sediment from a typical mariculture zone.

Seaweeds are good bio-monitors of heavy metals pollution in coastal seawater. In the present study, the potential bioremediation effects of cultivated Gracilaria lemaneiformis on heavy metals in Nan'ao coastal sediment from a typical mariculture zone, South China were evaluated. Sediment samples were collected from five different zones (Gracilaria cultivation zone, G; Fish culture zone, F; Shellfish culture zone, S; Transition zone, T; Control zone, C) from December 2014 to July 2015. The concentrations of Cd, Pb, Cu, and Zn in the sediments were significantly different among the various types of mariculture areas. The concentrations varied widely: Cd (0.04-1.02) µg g-1; Cu (1.19-37.70) µg g-1; Pb (8.45-74.45) µg g-1; Zn (36.80-201.24) µg g-1. The lowest heavy metal concentrations in the sediment were occurred at Gracilaria cultivation zone, while higher concentrations occurred at control zones and fish culture zones. The pollution load index, principal components and cluster analysis showed that heavy metal concentrations were the highest at fish culture zone, while the concentrations were the lowest at Gracilaria cultivation zone, and Gracilaria cultivation affects the heavy metals in the sediments. Gracilaria had strong adsorption capacities for heavy metals from seawater, showing the highest heavy metal Bioconcentration Factors in May (higher seaweed biomass period). Consequently, the results suggested that Gracilaria cultivation influences the heavy metal concentrations in sediments from the typical coastal mariculture zone. Gracilaria cultivation has the potential to bioremediate heavy metals in the coastal sediments. Therefore, Gracilaria cultivation can add environmental advantages and ecological values to coastal mariculture zones.

The anti-aging effects of Gracilaria lemaneiformis polysaccharide in Caenorhabditis elegans.

The anti-aging activity of marine macroalgae Gracilaria lemaneiformis polysaccharide (GP) on Caenorhabditis elegans was evaluated by observing the lifespan, reproduction, pharyngeal pumping and stress response of worms. Moreover, quantitative fluorescence of polyglutamic acid and nuclear localization of DAF-16 were observed. The results showed that GP treatment enhanced the mean lifespan by over 16.47% and significantly increased the reproduction duration of worm in the high dose group (1000 µg/mL). GP exhibited little potent effects under the thermotolerance and oxidative stress. The number of polyglutamic acid aggregates in three dosage groups decreased by 24.82%, 32.08% and 30.93% (p < 0.05) compared to the control. The middle dose group strongly induced DAF-16 nuclear translocation over intermediate and cytosolic localizations compared to the control (p < 0.001). It was inferred that GP extended the adult lifespan of wild-type and polyQ nematodes through the insulin pathway DAF-16.

Prostaglandins in Marine Organisms: A Review.

Prostaglandins (PGs) are lipid mediators belonging to the eicosanoid family. PGs were first discovered in mammals where they are key players in a great variety of physiological and pathological processes, for instance muscle and blood vessel tone regulation, inflammation, signaling, hemostasis, reproduction, and sleep-wake regulation. These molecules have successively been discovered in lower organisms, including marine invertebrates in which they play similar roles to those in mammals, being involved in the control of oogenesis and spermatogenesis, ion transport, and defense. Prostaglandins have also been found in some marine macroalgae of the genera Gracilaria and Laminaria and very recently the PGs pathway has been identified for the first time in some species of marine microalgae. In this review we report on the occurrence of prostaglandins in the marine environment and discuss the anti-inflammatory role of these molecules.

An Alternative Pathway to Leukotriene B4 Enantiomers Involving a 1,8-Diol-Forming Reaction of an Algal Oxylipin.

Oxidized lipids function as tissue hormones in mammals. An important class of these oxylipins are the immunomodulatory leukotrienes (LTs). Besides mammals, marine algae produce bioactive oxylipins, including LTs. The novel acid-labile oxylipin, (5 R,8 S)-dihydroxy eicosatetraenoic acid, from the edible alga Gracilaria vermiculophylla rearranges via a 1,8-diol-forming mechanism to inflammatory LTB4 enantiomers that exhibit an enantio-specific potency toward LTB4 receptor 1. This alternative pathway to a well-known leukotriene may explain food poisoning after Gracilaria consumption.

Toxicity and Mechanisms of Action of Polycyclic Aromatic Hydrocarbon Pollution in Red Algae (Gracilaria corticata) from the Northern Coast of the Oman Sea.

The activities of selected biomarkers including 7-ethoxycoumarin-O-deethylase (ECOD) and glutathione S-transferase (GST) were measured in the red algae (Gracilaria corticata) obtained from the Oman Sea. Chemical analyses were used to assess the polycyclic aromatic hydrocarbons (PAHs) in the red algae. Total PAH concentrations in the red algae ranged from 3.61 to 8.14 ng g-1 dry weight. Mean GST and ECOD activity also varied from 8.87 to 15.32 nmol/mg protein/min and from 0.31 to 1.02 pmol/min/mg protein, respectively. Significant correlations were found between the total PAH levels and the selected biomarkers (p < 0.01). The results showed that ECOD and GST enzymes reacted to PAHs in phase I and II detoxification mechanisms of red algae (G. corticata), which extend the use of these biomarkers for investigation of the biological effects of PAH pollution as well as determination of pollution bioavailability. Environ Toxicol Chem 2019;38:1947-1953. © 2019 SETAC.

A sulfated polysaccharide from Gracilaria Lemaneiformis regulates cholesterol and bile acid metabolism in high-fat diet mice.

This study aimed to evaluate the regulation of lipid metabolism and mechanism of a sulfated polysaccharide from Gracilaria Lemaneiformis (GLP). The results showed that GLP significantly decreased the hepatic total cholesterol (TC) and triglyceride (TG) levels in the serum of high-fat diet mice and prevented body weight gain and alleviated the accumulation of fat. Furthermore, GLP may accelerate the conversion of cholesterol to bile acids by promoting the expression of LxRα and CYP7A1 genes and regulating the relative abundance of Bacteroides, Ruminococcus_1 and Lactobacillus. Pearson's correlation analysis showed that the relative abundance of Alistipes, Prevotellaceae_UCG-001, and Corprococcus_1 was negatively associated with the hydrophobic BAs CDCA and DCA levels, while the relative abundance of Lachnospiraceae_NK4A136_group and Roseburia was positively correlated with the hydrophilic BAs UDCA and TUDCA levels. This study suggests that GLP can be used as a functional food for regulating cholesterol metabolism.

Anti-angiogenic activity of Gracilaria coronopifolia J.G. Agardh extract by lowering the levels of trace metals (iron, zinc and copper) in duck chorioallantoic membrane and in vitro activation of AMP-kinase.

AMP-activated protein kinase (AMPK) is an intracellular energy sensor important in metabolic regulation, cell growth, and survival. However, the specific role of AMPK signaling pathway in the inhibition of angiogenesis remains unclear. The study highlights the activity on AMP activated protein kinase signaling pathways of a marine algae, Gracilaria coronopifolia, and its effects on angiogenesis. It was found that the most potent extract, GCD, inhibited angiogenesis significantly in the duck chorioallantoic membrane assay and also activated the enzyme AMP-kinase, in vitro. The dichloromethane extract was found most active in inhibiting angiogenesis in the duck chorioallantoic membrane (IC50 = 1.21 µg/mL) followed by GCH (IC50 = 3.08 µg/mL) (p = 0.479) and GCM (IC50 = 8.93 µg/mL) (p = 0.042). Benferroni post hoc analysis revealed that there was no significant difference between the percent inhibitions of GCH and GCM extracts (p = 0.479). Consequently, angiogenic inhibition caused lowering of iron, zinc, and copper levels in the duck CAM. Thin layer chromatography and gas chromatography-mass spectrometry revealed the components of each extracts. Notably, this is the first report on the kinase activity of a red algae G. coronopifolia extracts and a colorimetric-based quantification of angiogenesis based on metal content of CAM. Our data also suggest a novel therapeutic approach for inhibiting angiogenesis through the AMPK pathway.