Red Alga Gracilaria Verrucosa Research Articles

Characterization of red algae (Gracilaria verrucosa) on potential application for topical treatment of oral mucosa wounds in Rattus norvegicus.

Wound healing in the mouth has its challenges due to masticatory movements and the presence of bacteria that can inhibit its process. The aim of this study was to analyze the contents of red algae (Gracilaria verrucosa) from Indonesia, and its potential as a wound-healing agent for oral wounds using animal model. Red algae content was determined by phytochemical tests and gas chromatography-mass spectroscopy (GC-MS). The wound was made by making an incision on the gingival mucosa of Rattus norvegicus and the parameters assessed were bleeding time, number of fibroblast cells, and time of wound closure. Three doses of G. verrucosa gel were used (2.5%, 5%, and 10%) and the gels were applied twice a day, at 6:00 and 18:00. Application was carried out topically by applying 0.1 ml of gel to the incision wound using a 1 mL syringe. Our phytochemical test indicated that the G. verrucosa contained alkaloids, steroids, flavonoids, and phenols. The dominant contains of the G. verrucosa were glycerol (36.81%), hexadecenoic acid (20.74%), and cholesterol (7.4%). The hemostasis test showed that the 2.5% G. verrucosa extract gel had the shortest bleeding time, 33.98±2.66 seconds. On the seventh day of the initial proliferation phase, the number of fibroblasts was not significantly different among groups. On day 14, the number of fibroblasts was only significantly different between 10% G. verrucosa and untreated group (p=0.007). On day 28, however, both 5% and 10% G. verrucosa were significantly higher compared to untreated group, both had p=0.010. Daily clinical examination showed that animals that were given 2.5% and 5% of G. verrucosa extract gel experienced wound closure on day 10. Animals treated with 10% of extract gel, all wounds healed on day 9. This study suggested that G. verrucosa extract could accelerate wound closure and wound healing.

Cyclopentenone and wickerol derivatives from the marine algicolous fungus Trichoderma atroviride A-YMD-9-4

A new cyclopentenone derivative, 4-hydroxyhypocrenone A, and a new naturally-occurring wickerol derivative, 8-acetoxywickerol A, as well as two known compounds, hypocrenone A and wickerol B, were purified from Trichoderma atroviride A-YMD-9-4, an endophytic fungus isolated from the marine red alga Gracilaria verrucosa. The structures and relative configurations of two new isolates were established by a combination of 1 D/2D NMR, IR, and mass spectroscopic methods, and the absolute configuration of 1 was assigned on the basis of ECD data analyzed by quantum chemical calculations. Compounds 1 and 2 exhibited weak inhibition of one or two marine phytoplankton species.

Harziane and cadinane terpenoids from the alga-endophytic fungus Trichoderma asperellum A-YMD-9-2

In addition to the known harziandione (1), one new diterpenoid, 3S-hydroxyharzianone (2), and two new sesquiterpenoids, 4-cadinen-11,12-diol (3) and 4-cadinen-11,13-diol (4), were isolated from the culture of Trichoderma asperellum A-YMD-9-2, an endophytic fungal strain from the marine red alga Gracilaria verrucosa. The structures and relative configurations of new isolates were established by analysis of spectroscopic techniques, including 1D/2D NMR, IR, and MS, and their absolute configurations, except for those at C-11 of 3 and 4, were assigned by comparison of the ECD curve or specific optical rotation data with those of analogues. Compound 2 may be an intermediate in the biosynthesis of harziandione from harzianone, while 3 and 4 represent the second occurrence of cadinane sesquiterpenoids in Trichoderma. Compounds 2−4 exhibited potent inhibition of several marine phytoplankton species.

Antibacterial activity of Caulerpa racemosa against pathogenic bacteria promoting “ice-ice” disease in the red alga Gracilaria verrucosa

Increasing ocean temperatures associated with climate change have triggered the occurrence of diseases in marine resources such as macroalgae or seaweed. “Ice-ice” is one of the most devastating diseases affecting economically important seaweeds such as Gracilaria and Eucheuma. In this study, we investigate the bacterial composition of diseased and healthy Gracilaria verrucosa, a red seaweed cultured in brackish water ponds in Takalar, Indonesia. Morphologic and phenotypic characteristics showed that the isolates from diseased Gracilaria belong to various genera: Vibrio, Chromobacterium, Flavobacterium, Pseudomonas, and Achromobacter. Several bacteria were also isolated from healthy Gracilaria including Corynebacterium, Serratia, Shigella, Micrococcus, Proteus, and Flavobacterium. Using Koch’s postulates, bacterial pathogenicity was established by bath exposure of naive G. verrucosa to each of the bacteria isolated from diseased Gracilaria resulting in symptom characteristic of “ice-ice” disease. The antibacterial property of the green seaweed Caulerpa racemosa against the pathogenic bacteria was assessed using extracts that were prepared with solvents of various polarities such as hexane, chloroform, ethyl acetate, methanol, and methanol-water. The highest antibacterial activity was observed in methanol extracts Caulerpa while extracts using the other solvents showed moderate to low activities. These findings demonstrate the potential of Caulerpa to inactivate bacterial pathogens associated with “ice-ice” disease.

Seven chromanoid norbisabolane derivatives from the marine-alga-endophytic fungus Trichoderma asperellum A-YMD-9-2

An examination of the endophytic fungus Trichoderma asperellum A-YMD-9-2 obtained from the marine red alga Gracilaria verrucosa led to the isolation of seven new chromanoid norbisabolane derivatives, trichobisabolins I–L (1–4) and trichaspsides C–E (5–7). Their structures and relative configurations were established on the basis of spectroscopic techniques, mainly including 1D/2D NMR and MS, and the absolute configuration of 1 was assigned by X-ray crystallographic analysis using Cu Kα radiation. All of these isolates feature a 1,9-epoxy ring system, and 5–7 represent the second occurrence of norbisabolane aminoglycosides. Compounds 1–7 exhibited potent inhibition of several marine phytoplankton species.

Cyclonerane Derivatives from the Algicolous Endophytic Fungus Trichoderma asperellum A-YMD-9-2.

Seven previously unreported cyclonerane derivatives, namely, 3,7,11-trihydroxycycloneran-10-one, cycloneran-3,7,10,11-tetraol, cycloneran-3,7,11-triol, 11,12,15-trinorcycloneran-3,7,10-triol, 7,10S-epoxycycloneran-3,15-diol, 7,10R-epoxycycloneran-3,15-diol, and (10Z)-15-acetoxy-10-cycloneren-3,7-diol, were isolated in addition to the known (10Z)-cyclonerotriol, (10E)-cyclonerotriol, catenioblin C, and chokol E from the culture of Trichoderma asperellum A-YMD-9-2, an endophytic fungus obtained from the marine red alga Gracilaria verrucosa. The structures of previously unreported compounds were established by spectroscopic techniques, including 1D/2D NMR, MS, and IR. The isolation of these new cyclonerane derivatives greatly adds to the structural diversity of unusual cyclonerane sesquiterpenes, and several isolates exhibit potent inhibition against some marine phytoplankton species.

Combined Effect of Icing Medium and Red Alga Gracilaria verrucosa on Shelf Life Extension of Indian Mackerel (Rastrelliger kanagurta)

Flake ice refrigeration is a conventional but commonly employed technique to slow down fish spoilage and deterioration of nutritional values and sensory properties during chilled storage. In the present study, a methanolic extract of red alga Gracilaria verrucosa was characterised, and functional groups, such as alkenes, aldehydes, nitriles, galactans and galactose-4-sulphate were found. Subsequent identification of active compounds revealed the presence of potent preservative agents, such as butylated hydroxytoluene, sulfurous acid, 1,2-propanediol, benzeneacetic acid, cyclononasiloxane and tetracosamethyl-cyclododecasiloxane. The effect of incorporating G. verrucosa at two different concentrations (0.67 and 2.5 g lyophilised alga/L aqueous solution) in the icing medium was tested for the preservation of Indian mackerel (Rastrelliger kanagurta), compared to preservation in traditional ice prepared only from water. Microbial, chemical and sensory qualities were monitored in Indian mackerel chilled in ice with and without G. verrucosa during 15 days storage period. Inhibitory effects (p < 0.05) on the microbial proliferation (mesophilic and psychrophilic bacteria) and chemical markers of fish deterioration (pH, TVB-N, TMA-N and biogenic amines) were evidenced for ice containing both concentrations of G. verrucosa, respectively, relative to the control medium. The sensory score acceptability limit reached 11 days for Indian mackerel stored in traditional ice and 15 days for Indian mackerel stored in ice with G. verrucosa extract. Thus, the icing medium containing G. verrucosa extract improves the quality and safety of Indian mackerel during storage and can be explored by the seafood industry as a biopreservative.

Agar yield from R-phycoerythrin extraction by-product of the red alga Gracilaria verrucosa

R-phycoerythrin (RPE) extracted from the red alga Gracilaria verrucosa by enzymatic hydrolysis generates a large amount of coproduct containing agar. We developed a process for the sequential extraction of RPE and agar. A Box–Behnken design with three factors–three levels and response surface methodology were applied to study the influence of different extraction parameters (incubation temperature, incubation time, and algae-to-water ratio) on the yield, 3,6-anhydrogalatose (3,6-AG) content, and sulfated degree of agar from RPE coproduct. The extraction efficiency was maximized (28%) when the extraction process was carried out with these conditions: incubation temperature of 100 °C, time of 2.6 h, and algae-to-water ratio equal to 3 (% w/v). Optimum 3,6-AG content (30%) was maximized at 85 °C, 1.85 h, and 2.35 (% w/v). Sulfated degree varied between 6 and 12%. The results of this study indicate that fresh G. verrucosa can be subjected to a process of simultaneous extraction of R-phycoerythrin and agar.

Antibacterial activity of red algae (Gracilaria verrucosa) extract against Escherichia coli and Salmonella typhimurium

Red alga was widely used in several fields, including food, feed, phamacy and industrial point of view. The chemical analysis showed that red alga contained terpenoid, acetogenic, and aromatic compounds, which have a wide range of biological activities, such as anti-micobial, anti-inflammatory and anti-viral. The objectives of this research was to evaluate the effect of extraction solvent and time on antibacterial activity of red alga (Gracilaria verrucosa), and to explore the bioactive compound contained within Gracilaria verrucosa. The method in this study used descriptive reseach. These findings revealed that the highest inhibition activity among all extracts was obtained with the ratio of methanol:aquades (75:25) and extraction time around 72 hours against Escherichia coli and Salmonella typhimurium. The bioactive compounds of Gracilaria verrucosa tested by phytochemical analysisi consisted of flavonoid, alkaloid, and saponin. Those secondary metabolites may be approximated as antibactial substances.

해조류 바이오매스로부터 바이오슈거 생산 - 열수 및 냉동 전처리의 영향

In this study, the production of reducing sugar from red-algae Gracilaria verrucosa was investigated using hotwater and freezing pretreatments and subsequent enzymatic hydrolysis. In hot-water pretreatment, 2.16% total reducing sugar (TRS) yield (3.62% based on carbohydrate) was obtained under the condition of 130ºC and 120 min. By subsequent enzymatic hydrolysis, 27.9% TRS yield (46.8% based on carbohydrate) was achieved. In the freezing pretreatment, about 20% TRS yields were similarly obtained in the range of -70℃ to -20℃. Also, solid-to-liquid ratio of 1:10-1:15 presented the similar TRS yield. Moreover, freezing time did not affect TRS yield within tested range of 48 hr. In freezing pretreatment, 3.15% TRS yield (5.28% based on carbohydrate) was obtained under the condition of -70℃ and 48 hr. By subsequent enzymatic hydrolysis, 19.5% TRS yield (32.7% based on carbohydrate) was achieved. From these results, hot-water and freezing pretreatments are available to produce biosugar from G. verrucosa for bio-refinery process.

Pengaruh Pemberian Ekstrak Alga Merah (gracilaria verrucosa) terhadap Aktivitas SGPT pada Tikus Putih (rattus norvegicus) Jantan Galur Wistar yang Diinduksi Parasetamol Dosis Tinggi

Background: Administration of high dose paracetamol may increase the metabolic pathways that produce N-acetyl-p-benzoquinonimin (NAPQI). NAPQI is a reactive substance, resulting liver tissue damage and increase SGPT activity. Red algae extract contains flavonoids that act as antioxidants.Objectives: This research aimed to determine the effect of red algae (Gracilaria verrucosa) extract to the activity of serum glutamic pyruvic transaminase (SGPT) in male Wistar rats (Rattus norvegicus) induced by high dose paracetamol.Methodology: This research used 24 animal of Male Wistar rats. These rats were divided into 3 groups: 1) group of rats fed with standard food; 2) group of rats given paracetamol 1750 mg/kg on day 10; and 3) group of rats given red algae extract 200 mg/kg/day on day 1-11 and paracetamol 1750mg/kg on day 10. On day 11, all rats were sacrificed and SGPT activities were measured by spectrophotometry method.Result: Mann-Whitney test results showed significant difference (p = 0.021) between SGPT activity of group of rats fed with standard food (=81.95±10.103 U/L) and group of rats induced by high dose paracetamol (=97.06±11.19 U/L). Furthermore, there was significant difference (p = 0.016) between SGPT activity of group of rats induced by high dose paracetamol (=97.06±11.19 U/L) and group of rats induced by high doses of paracetamol and fed with red algae extract (=82.73±11.233 U/L).Conclusion: The conclusion of this research showed that high dose paracetamol significantly increased SGPT activity and red algae extract significantly decreased SGPT activity because it contains flavonoids that act as antioxidants.

Production of sugars from macro-algae Gracilaria verrucosa using combined process of citric acid-catalyzed pretreatment and enzymatic hydrolysis

In this study, a combined process of citric acid-catalyzed pretreatment and enzymatic hydrolysis was applied for production of total reducing sugar (TRS) from marine macro red-algae Gracilaria verrucosa as a potential resource. By citric acid-catalyzed pretreatment under the 1:10 solid-to-liquid ratio, 0.1M citric acid, 150°C, 60min condition, a 50.9% TRS yield was obtained. By subsequent enzymatic hydrolysis using enzyme mixture (20 FBG/g-biomass and 28 FPU/g-biomass), a 57.8% TRS yield was obtained. Because citric acid-catalyzed pretreatment is simple, cheap, and environmentally friendly (do not use hazardous chemicals), these results served to demonstrate the significant potential of citric acid-catalyzed pretreatment to produce TRS for biofuels and biochemicals refinement purposes.

Conversion of red-algae Gracilaria verrucosa to sugars, levulinic acid and 5-hydroxymethylfurfural.

This study employed a statistical methodology to investigate the optimization of conversion conditions and evaluate the reciprocal interaction of reaction factors related to the process of red-algae Gracilaria verrucosa conversion to sugars (glucose, galactose), levulinic acid and 5-hydroxymethylfurfural (5-HMF) by acidic hydrolysis. Overall, the conditions optimized for glucose formation included a higher catalyst concentration than did those for galactose, and these conditions for galactose were similar to those for 5-HMF. Levulinic acid production, meanwhile, was optimized at a higher reaction temperature, a higher catalyst concentration, and a longer reaction time than was glucose, galactose or 5-HMF production. By this approach, the optimal yields (and reaction conditions) for glucose, galactose, levulinic acid, and 5-HMF were as follows: glucose 5.29 g/L (8.46 wt%) (reaction temperature 160 °C, catalyst concentration 1.92%, reaction time 20 min), galactose 18.38 g/L (29.4 wt%) (160 °C, 1.03%, 20 min), levulinic acid 14.65 g/L (18.64 wt%) (180.9 °C, 2.85%, 50 min), and 5-HMF 3.74 g/L (5.98 wt%) (160.5 °C, 1%, 20 min).

Resultados preliminares del cultivo experimental de Gracilaria verrucosa (Hudson) Papenfuss (=G. caudata J. Agardh) (Rhodophyta: Gracilariaceae) en la Costa Caribe de Colombia

We present the results of the study carried out in Fray Domingo Bay, Punta de Piedra Beach (10° 52´ N and 75° 08´ W approximately), Department of Atlántico, on the Colombian Caribbean coast, from June 30 to November 1, 2004. An experimental crop of the red algae Gracilaria verrucosa was developed using a handmade structure whose main rope with plastic perforated sleeves served as support for the algae. This structure was placed at a depth of 1.40 m and fixed to the bottom to maintain it floating. The structure was inspected every 15 days to observe the development of new branches and the apical growth in the filaments. The growth of the algae was irregular, as the feathers of those originally fixed grew evenly while in others growth was observed only in the end sections of the sleeves. The fresh weight of the main rope with sleeves and algae was 2.5 kg. The longest filaments were 22 cm long. Fronds growth average was of 15 cm. Cultivated algae grew without invasive attacks, but we did observe abundant epiphytes on the main and secondary ropes of the structure, which belonged to the Rhodophyceae, Phaeophyceae and Clorophyceae classes in order of abundance. Hypnea musciformis was the most abundant species and showed the best growth rate. The associated fauna consisted mainly of juvenile crustaceans: crabs, shrimps and lobsters.

Oxidative Stress and Antioxidant Activities of Intertidal Macroalgae in Korea

The oxidative stress level and antioxidant activities in two green algae (Ulva pertusa and Ulva linza), two brown algae (Agarum cribrosum and Dictyota dichotoma), and three red algae (Grateloupia lanceolata, Carpopeltis affinis, and Gracilaria verrucosa) collected from intertidal regions of Korea were assessed. In the two green algae, although the total glutathione content was not as high as that of the brown algae, the glutathione pool was extremely reduced, and the glutathione reductase (GRd)/glutathione peroxidase (GPx) activity ratio was high, which apparently plays an important role for protection against oxidative damage, as manifested by low lipid peroxidation. In the brown algae, which exhibited a low lipid peroxidation level that was comparable to the green algal species, the highest glutathione content, together with high GPx activity, appears to be the most important factor in their antioxidant protection. The red algal species exhibited extremely high lipid peroxidation levels. They also contained the lowest and most oxidized glutathione among the species, as well as the lowest GRd activity. In spite of the marked difference in the glutathione content, the significant difference in the activity of γ-glutamylcysteine ligase, the rate limiting enzyme for glutathione synthesis, among the species was not exhibited. Our results suggest that there is a significant difference in the levels of oxidative stress and antioxidant capacity among the algal species, and that the glutathione system, especially the efficiency of glutathione recycling, plays a vital role in antioxidative protection in algal species.

A statistical approach for optimization of R-phycoerythrin extraction from the red algae Gracilaria verrucosa by enzymatic hydrolysis using central composite design and desirability function

The aim of this research is to statistically optimize enzymatic hydrolysis parameters for the production of R-phycoerythrin (RPE) from red algae Gracilaria verrucosa. Six independent variables, incubation temperature, incubation time, ratio of buffer to raw material, cellulase loading, xylanase loading, and pH, were selected for response surface methodology studies. A central composite design was employed to maximize RPE production. A mathematical model with high determination coefficient (R 2 = 0.86) was developed and could be employed to optimize RPE extraction. The optimal extraction conditions of RPE were determined as follows: incubation temperature (48°C), incubation time (6 h), ratio of buffer to raw material (20 w/v), cellulase loading (15%), xylanase loading (5%), and pH (6.5). Under this optimal condition, the experimental yield of RPE was 6.25 mg g−1. Based on the result of response surface methodology and desirability function approach study, total sugar, the main by-product in RPE extraction was considered as another response. A new optimal condition was predicted as follows: incubation temperature (30°C), incubation time (12 h), ratio of buffer to raw material (20, w/v), cellulase loading (15%), xylanase loading (5%), and pH (6). Under this condition, similar RPE levels were obtained while the concentration of total sugar decreased by 40%.

A natural anti-inflammatory enone fatty acid inhibits angiogenesis by attenuating nuclear factor-κB signaling in vascular endothelial cells

An anti-inflammatory enone fatty acid, (E)-9-oxooctadec-10-enoic acid (C10), was previously isolated from red alga (Gracilaria verrucosa). Of the many cellular signaling pathways activated in response to the inflammatory stimulus, lipopolysaccharide, the extracellular signal-regulated kinase 1/2, the stress-activated protein kinase/Jun N-terminal kinase and the nuclear factor-κB pathways were specifically blocked by C10 in the macrophage-like cell line, RAW264.7. In this study, we investigated the anti-angiogenic and anti-inflammatory activities of C10 in endothelial cells. C10 only partially inhibited the proliferation of human cancer cell lines at relatively high concentrations of over 20 μg/ml. However, C10 inhibited the proliferation of RAW264.7 cells and human umbilical vein endothelial cells (HUVECs) with half-maximal inhibitory concentration (IC50) values of 4-8 μg/ml. Both the proliferation and the migration of HUVECs induced by the vascular endothelial growth factor (VEGF) were markedly blocked by C10 with IC50 values of 2-3 μg/ml. The activation of nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor, α, by tumor necrosis factor-α or VEGF in these cells was also blocked by C10. Furthermore, in an in vivo model of angiogenesis in the mouse cornea, the neovascularization induced by VEGF was markedly inhibited by C10. The processes involved in inflammatory signaling, angiogenesis, and the development of malignancy in cancer are closely related, suggesting that C10 could be a useful lead compound for the development of novel anti-angiogenic therapies for cancer.

Cultivation of Gracilaria verrucosa (Huds) Papenfuss in Chilika Lake for livelihood generation in coastal areas of Orissa State

The agarophyte red alga Gracilaria verrucosa occurs widely in Chilika Lake, one of the RAMSAR wetland sites in India. The lake is situated in the extreme southeast corner of Orissa between latitudes 19°28′ and 19°54′ N and longitudes 85°06′ and 85°35′ E. The natural biomass production is not sufficient for the agar industry, and the only alternative is to maximize the production of the seaweed through mass cultivation by seaweed farming. To elucidate important aspects of the growth and development of G. verrucosa, experimental field cultivation was undertaken at Langaleswar and Samal sites of Chilika Lake using ropes and raft methods during March to August, 2009. After 30 days of cultivation a maximum 15- and 13.8-fold increase in biomass in raft culture and rope culture, respectively, was observed at Langaleswar and an 11.6- and 11.0-fold increase in biomass at Samal. Environmental parameters such as temperature, salinity, pH, transparency, DO, conductivity, nitrate, and phosphate were monitored at both stations, and the influence of environmental parameters is discussed.

Studies on chemical constituents of Gracilaria verrucosa

A new sterol glucoside, (24R)-5α-stigmast-9-(11)-en-3β-D-glucopyranoside (1) has been isolated from a methanol extract of the red alga Gracilaria verrucosa, while cholesterol and (Z)-9-hexadecenoic acid (palmitoleic acid) have been isolated from a dichloromethane–acetone extract of the same alga. Additionally, an oily fraction of a dichloromethane–acetone extract has been investigated by gas chromatography/mass spectrometry (GC/MS). The main components in the oily fraction were heneicosanoic acid (49.8%), (Z)-9-octadecenoic acid (15.8%), and 15-tetracosenoic acid (15.6%) in addition to a few hydrocarbons, simple aromatics and a new compound, namely 3,4-dimethyl-5-carboxyclic acid-1-oxacyclopent-3-en-2-one. The structures of the isolated compounds have been elucidated by spectroscopic methods.

Anti-inflammatory Constituents of the Red Alga Gracilaria verrucosa and Their Synthetic Analogues

A chemical study on the anti-inflammatory components of the red alga Gracilaria verrucosa led to the isolation of new 11-deoxyprostaglandins ( 1- 4), a ceramide ( 5), and a C 16 keto fatty acid ( 6), along with known oxygenated fatty acids ( 7- 14). Their structures were elucidated on the basis of NMR and MS data. The absolute configurations of compounds 1- 5 were determined by Mosher's method. The anti-inflammatory activity of the isolated compounds ( 1- 14) was evaluated by determining their inhibitory effects on the production of pro-inflammatory mediators (NO, IL-6, and TNF-alpha) in lipopolysaccharide (LPS)-activated RAW 264.7 murine macrophage cells. Compounds 9 and 10 exhibited the most potent activity. In the evaluation of these two compounds and derivatized analogues ( 15- 40), the anti-inflammatory activity was enhanced in some synthetic analogues. These enone fatty acids were investigated as potential anti-inflammatory leads for the first time.