Gelidium Amansii Research Articles

Ethanol Production from the Seaweed Gelidium amansii, Using Specific Sugar Acclimated Yeasts

For the production of ethanol from seaweed as the source material, thermal acid hydrolysis and enzymatic saccharification were carried out for monosugars production of 25.5 g/l galactose and 7.6 g/l glucose using Gelidium amansii. The fermentation was performed with Pichia stipitis KCTC 7228 or Saccharomyces cerevisiae KCCM 1129. When wild P. stipitis and S. cerevisiae were used, the ethanol productions of 11.2 g/l and 6.9 g/l were produced, respectively. The ethanol productions of 16.6 g/l and 14.6 g/l were produced using P. stipitis and S. cerevisiae acclimated to high concentration of galactose, respectively. The yields of ethanol fermentation increased to 0.5 and 0.44 from 0.34 and 0.21 using acclimated P. stipitis and S. cerevisiae, respectively. Therefore, acclimation of yeasts to a specific sugar such as galactose reduced the glucose-induced repression on the transport of galactose.

해조류 우뭇가사리 (Gelidium amansii)의 분리당화발효를 이용한 바이오에탄올의 생산

The seaweed, Gelidium amansii, was fermented to produce bioethanol. Optimal pretreatment condition was determined as 94 mM H₂SO₄ and 8% (w/v) seaweed slurry at 121℃ for 60 min. The mono sugars of 40.4 g/L with 67% of conversion from total carbohydrate of 60.6 g/L with 80 g dw/L G. amansii slurry were obtained by thermal acid hydrolysis pretreatment and enzymatic saccharification. G. amansii hydrolysate was used as the substrate for ethanol production by Kluyveromyces marxianus KCTC 7150 and Candida tropicalis KCTC 7212 using 5L fermentor. The ethanol productions by K. marxianus KCTC 7150 and C. tropicalis KCTC 7212 were 17.8 g/L with Y EtOH of 0.48 at 120 h and 19.3 g/L with Y EtOH of 0.50 at 120 h, respectively.

Production of mixed acids from non-pretreated red algae Gelidium amansii

The present study investigates the production of mixed acids by anaerobic fermentation using non-pretreated red algae Gelidium amansii as a substrate. The production levels of mixed acids were increased 1.6-fold in enrichment cultures of anaerobic digester sludge and tidal field soil compared with fresh sludge. The enrichment culture was composed of four major groups of microbial strains belonging to Clostridium sp. and Enterococcus sp. A defined mixed culture established based on the identified isolates increased mixed acids production level by up to 50.9% (g/g). Semi-continuous fermentation with a biomass-loading rate of 2.77 g/L/d and hydraulic retention time of 30 days yielded a productivity of 1.41 g/L/d. This is the first report of the production of mixed acids from the red algae G. amansii.

Immunomodulatory activities of Gelidium amansii gel extracts on murine RAW 264.7 macrophages

The objective of this study was to investigate the effects of Gelidium amansii, a red algae cultivated in the northeastern coast of Taiwan, on immune regulatory activities in RAW 264.7 macrophages. G. amansii gel was prepared by boiling and filtering the dried G. amansii algae. Phosphate-buffered saline (PBS) and ethanol extracts of the freeze-dried gel were then used to treat the cells. The results showed that the PBS extracts of G. amansii gel activated the macrophage by increasing cell proliferation and by enhancing the production of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6; the ethanol extracts did not show such effects. By contrast, neither PBS nor ethanol extracts of G. amansii gel affected NO production in lipopolysaccharide (LPS)-stimulated macrophages, although the ethanol extracts suppressed LPS-stimulated TNF-α, IL-1β, and IL-6 production. In conclusion, the PBS extracts of G. amansii gel activated the macrophage by enhancing the production of immune mediators, whereas the ethanol extracts showed anti-inflammatory activities by suppressing the cytokine production in LPS-stimulated RAW 264.7 macrophages.

우뭇가사리로부터 레불린산 생산공정을 위한 2단 산 가수분해

현재 1,2세대 바이오매스에 비해 상대적으로 값싸며 대량생산이 가능한 3세대 바이오매스인 해조류를 이용하여 다양한 바이오화합물 생산에 관한 연구들이 주목받고 있다. 이러한 이유는 해조류는 다른 바이오매스에 비해 빨리 자라나고, 큰 장비 없이도 쉽게 수확할 수 있다는 장점뿐만 아니라 다양한 화합물로 전환할 수 있는 당이 풍부하고 공정을 통해 쉽게 전환할 수 있기 때문이다. 이러한 해조류부터 다양한 바이오화합물을 생산하는데 있어서 한 가지로 resins, plasticizers, textiles, animal feed, coatings, antifreeze의 상업화된 공정에 사용할 수 있는 레불린산(levulinic acid)이 있다. 본 연구에서는 해조류로부터 효과적으로 레불린산을 생산하는데 있어서 온도, 시간, 산의 농도의 실험조건과 2단 산 처리 공정(two step acid treatment)을 통해 생산을 최적화 하는 조건을 탐색해 보았다. 첫번째 단계로는 상대적으로 저온에서 침지 공정을 통해 고상으로는 다양한 용도로 사용될 수 있는 셀룰로오스를 회수하고, 액상으로는 갈락토오스를 회수하였다. 2번째 단계로는 고온에서 회분식 공정을 통해 갈락토오스를 레불린산으로 전환하였다. 실험 결과 2단 산 처리 공정을 통해 초기바이오매스 기준 20.6%의 레불린산 수율을 확보하였다. The study of bioproduct production from inexpensive biomass such as marine biomass has recently attracted considerable attention. Because, marine biomass which compared to land biomass, it can be grown rapidly and is easily cultivated without the need for expensive equipment. In addition, the carbohydrate contents are similar or higher than land biomass such as woody biomass and can be easily converted to chemicals through proper chemical processes. In the production of various biochemicals from marine biomass, levulinic acid is a highly versatile chemical with numerous industrial uses and has the potential to become a commodity chemical. It can be used as a raw material for resins, plasticizers, textiles, animal feed, coatings and antifreeze. In this study, experiments were carried out to determine the optimum conditions of temperature, acid concentration and reaction time for production of levulinic acid from marine biomass, Gelidium amansii, using two-step treatment. In the first hydrolysis step, solid-state cellulose which was used to produce ethanol by fermentation and liquid-state galactose which used to produce bioproduct such as levulinic aicd were obtained through acid soaking. In the second hydrolysis step, the liquid-state galactose was converted into levulinic acid via a high-temperature reaction in a batch reactor. As a result, the overall production yield of Gelidium amansii to levulinic acid in the two-step acid hydrolysis was approximately 20.6% on the initial biomass basis.

Molecular Cloning, Overexpression, and Enzymatic Characterization of Glycosyl Hydrolase Family 16 β-Agarase from Marine Bacterium Saccharophagus sp. AG21 in Escherichia coli

An agar-degrading bacterium was isolated from red seaweed (Gelidium amansii) on a natural seawater agar plate, and identified as Saccharophagus sp. AG21. The β-agarase gene from Saccharophagus sp. AG21 (agy1) was screened by long and accurate (LA)-PCR. The predicted sequence has a 1,908 bp open reading frame encoding 636 amino acids (aa), and includes a glycosyl hydrolase family 16 (GH16) β-agarase module and two carbohydrate binding modules of family 6 (CBM6). The deduced aa sequence showed 93.7% and 84.9% similarity to β-agarase of Saccharophagus degradans and Microbulbifer agarilyticus, respectively. The mature agy1 was cloned and overexpressed as a His-tagged recombinant β-agarase (rAgy1) in Escherichia coli, and had a predicted molecular mass of 69 kDa and an isoelectric point of 4.5. rAgy1 showed optimum activity at 55oC and pH 7.6, and had a specific activity of 85 U/mg. The rAgy1 activity was enhanced by FeSO4 (40%), KCl (34%), and NaCl (34%), compared with the control. The newly identified rAgy1 is a β-agarase, which acts to degrade agarose to neoagarotetraose (NA4) and neoagarohexaose (NA6) and may be useful for applications in the cosmetics, food, bioethanol, and reagent industries.

Comparison of agarophytes (Gelidium, Gracilaria, and Gracilariopsis) as potential resources for bioethanol production

This study explores the possibility of producing ethanol using the acid hydrolysate of three abundant agar-containing red seaweeds (agarophytes): Gelidium amansii, Gracilaria tenuistipitata, and Gracilariopsis chorda. The main component in the seaweed samples was agar, which ranged from 20 to 51 % (g g−1 dry weight). After optimizing acid hydrolysis, 100 g of seaweed was hydrolyzed at 130 °C for 15 min with 0.2 M H2SO4. Then, 120 mL of a 1:2 mixture of the hydrolysate broth and basal medium was fermented in a 200-mL bottle at 30 °C for 96 h. Of the three seaweeds, G. amansii had the best ethanol yield, producing 0.23 g g−1 of galactose or 45 % of the theoretical yield. This yield increased to 60 % after detoxification of the hydrolysate with activated carbon.

Biotransformation of 5-hydroxymethylfurfural (HMF) by Scheffersomyces stipitis during ethanol fermentation of hydrolysate of the seaweed Gelidium amansii

The seaweed, Gelidium amansii, was fermented to produce bioethanol. Optimal pretreatment condition was determined as 94 mM H2SO4 and 10% (w/v) seaweed slurry at 121°C for 60 min. The mono sugars of 43.5 g/L with 57.4% of conversion from total carbohydrate of 75.8 g/L with G. amansii slurry 100g dcw/L were obtained by thermal acid hydrolysis pretreatment and enzymatic saccharification. G. amansii hydrolysate was used as the substrate for ethanol production by separate hydrolysis and fermentation (SHF). The ethanol concentration of 20.5 g/L was produced by Scheffersomyces stipitis KCTC 7228. The effect of HMF on ethanol production by S. stipitis KCTC 7228 was evaluated and 5-hydroxymethylfurfural (HMF) was converted to 2,5-bis-hydroxymethylfuran. The accumulated 2,5-bis-hydroxymethylfuran in the medium did not affect galactose and glucose uptakes and ethanol production. Biotransformation of HMF to less inhibitory compounds by S. stipitis KCTC 7228 could enhance overall fermentation yields of seaweed hydrolysates to ethanol.

Optimization of batch dilute-acid hydrolysis for biohydrogen production from red algal biomass

Marine algae are promising alternative sources for bioenergy including hydrogen. Their polymeric structure, however, requires a pretreatment such as dilute-acid hydrolysis prior to fermentation. This study aimed to optimize the control variables of batch dilute-acid hydrolysis for dark hydrogen fermentation of algal biomass. The powder of Gelidium amansii was hydrolyzed at temperatures of 120–180 °C, solid/liquid (S/L) ratios of 5–15% (w/v), and H2SO4 concentrations of 0.5–1.5% (w/w), and then fed to batch hydrogen fermentation. Among the three control variables, hydrolysis temperature was the most significant for hydrogen production as well as for hydrolysis efficiency. The maximum hydrogen production performance of 0.51 L H2/L/hr and 37.0 mL H2/g dry biomass was found at 161–164 °C hydrolysis temperature, 12.7–14.1% S/L ratio, and 0.50% H2SO4. The optimized dilute-acid hydrolysis would enhance the feasibility of the red algal biomass as a suitable substrate for hydrogen fermentation.

Optimization of levulinic acid production from Gelidium amansii

The study of bioproduct production, such as bioethanol and biochemicals, from inexpensive biomass has recently attracted considerable attention. Compared to land biomass, marine biomass can be grown rapidly and is easily cultivated without the need for expensive equipment. In addition, annual CO2 absorption by marine biomass is five to seven times higher than that of wood-biomass and the carbohydrate content is higher and can easily be converted to chemicals through proper chemical processes. In the production of various biochemicals from marine biomass, levulinic acid is a highly versatile chemical with numerous industrial uses and the potential to become a commodity chemical. It can be used as a raw material for resins, plasticizers, textiles, animal feed, coatings and antifreeze. The present study was carried out to determine the optimum conditions of temperature, acid concentration and reaction time for levulinic acid production from marine biomass Gelidium amansii using two-step treatment. In the first hydrolysis step, solid-state cellulose was obtained through acid soaking and used to produce ethanol by fermentation and liquid-state galactose. In the second hydrolysis step, the liquid-state galactose was converted into levulinic acid via a high-temperature reaction in a batch reactor. The overall production conversion of G. amansii biomass to levulinic acid in the two-step acid hydrolysis was approximately 20.6%.

Extract from Edible Red Seaweed (Gelidium amansii) Inhibits Lipid Accumulation and ROS Production during Differentiation in 3T3-L1 Cells

Gelidium (G.) amansii is a red alga widely distributed in the shallow waters around East Asian countries. We investigated the effect of G. amansii on lipid accumulation and ROS (Reactive Oxygen Species) production in 3T3-L1 cells. G. amansii extracts dose-dependently inhibited lipid formation and ROS generation in cultured cells. Our results showed that anti-adipogenic effect of G. amansii was due to the reduction in mRNA expressions of PPARγ peroxisome proliferator-activated receptor-γ and aP2 (adipocyte protein 2). G. amansii extracts significantly decreased mRNA levels of a ROS-generator, NOX4 (nicotinamide adenine dinucleotide phosphate hydrogen oxidase 4), and increased the protein levels of antioxidant enzymes including SOD1/2 (superoxide dis-mutases), Gpx (glutathione peroxidase), and GR (glutathione reductase), which can lead to the reduction of ROS in the cell. In addition, the G. amansii extract enhanced mRNA levels of adiponectin, one of the adipokines secreted from adipocytes, and GLUT4, glucose uptake protein. Taken together, our study shows that G. amansii extract inhibited lipid accumulation and ROS production by controlling adipogenic signals and ROS regulating genes.

The Marine AlgaGelidium amansiiPromotes the Development and Complexity of Neuronal Cytoarchitecture

Neurotrophic factors are vital not only to support neuronal development but also to protect mature neurons from atrophy in neurodegenerative diseases. As an effort to explore natural sources that possess neurotrophic activity, we screened common marine algae for their neuritogenic activity in the developing rat hippocampal neurons in culture. Of the 22 seaweed species examined, ethanol extracts of Gelidium amansii (GAE) exhibited potent neuritogenic activity, followed by Undaria pinnatifida and Sargassum fulvellum extracts. The effects of GAE were dose dependent with an optimum concentration of 15 µg/mL. The GAE significantly promoted the initial neuronal differentiation from the stage I into the stage II and increased the indices of axonal and dendritic development such as the length, the numbers of primary processes, and branching frequencies by a minimum of twofold compared with the vehicle control. These results show that marine algae are promising candidates for neurotrophic potentials.

Optimum separation condition of five bioactive compounds by HCI program in HPLC

Summary An efficient method used to separate five bioactive compounds from Gelidium amansii was optimized by the HCI software. The optimum composition of mobile phase for high-performance liquid chromatographic (HPLC) separation was obtained. The elution profiles were calculated by the polynomial theory based on the retention factor ln k = A + BF + CF2 (F was the volume percentage of acetonitrile with 1.0% acetic acid); then, the theory was applied to calculate the elution profile in both isocratic and gradient modes by modifying different mobile phase conditions with HCI program. The calculated results of mobile phase condition suggested that acetonitrile-water (containing 1.0% acetic acid) with a linear gradient elution of 0∼30 min from 15:85 to 50:50 (v/v) was the optimized component. In the experimental conditions, the agreement between the experimental elution profiles and the calculated values of eluted concentration was relatively good.

Production of mono sugar from acid hydrolysis of seaweed

The purpose of this work was to optimize the process conditions for the saccharification of macroalgae (seaweed) into mono sugar using the following parameters such as: Amount of biomass, catalyst concentration, temperature and reaction time. The major component of Ulva pertusa (green seaweed), Laminaria japonica (brown seaweed) and Gelidium amansii (red seaweed) is carbohydrate which can be converted into variety of sugars using sulfuric acid with hot-compress treatment. Rhamnose (37.89 wt %) and glucose (16.14 wt %) were extracted from dried U. pertusa , while galactose (49.32 wt %) and glucose (12.62 wt %) were extracted from dried G. amansii . Mannitol (31.53 wt %) was produced from dried L. japonica . Key words: Marine biomass, seaweed, saccharification, sulfuric acid hydrolysis, mono sugars.

Optimization of enzymatic extraction of polysaccharides from some marine algae by response surface methodology

A novel enzymatic extraction combined with response surface methodology was developed for polysaccharide extraction from marine algae. Box-Behnken design was employed to optimize concentration of enzyme, ratio of water to raw material and extraction time for high extraction yields. The optimal extraction conditions were as follows: a concentration of enzyme of 778.01 mg/g with ratio of water to raw material of 86.80 and extraction time of 129.93 min for Gelidium amansii and a concentration of enzyme of 997.03 mg/g with ratio of water to raw material of 98.76 and extraction time of 117.69 min for Laminaria japonica Aresch. Under the optimal conditions, extraction yields were 48.36% and 32.47%, respectively. Experimental data were fitted by multiple regression analysis to a secondorder polynomial equation and were statistically analyzed. The predicted model matched the experimental data well with coefficients of determination (R2) of 0.9864 and 0.9892, respectively.

Successful preparation and characterization of biotechnological grade agarose from indigenous Gelidium amansii of Taiwan

This paper reports the first successful preparation of biotechnological grade agarose from Gelidium amansii found in Taiwan. The scale-efficiency preparation was achieved by shortening EDTA treatment time through dispersing G. amansii agar in water in the presence of heat and EDTA, removing agaropectin impurity with a heat-compatible and strong-anion exchange resin, and precipitating agarose with a cost-effective isopropanol method. The yield of agarose from prepared G. amansii agar was 11.3%. The acquired agarose has a gel strength of 853gcm−2, a sulfate content of 0.14%, a pyruvate content of 1.03%, a degree of electroendosmosis of 0.16 and very limited binding affinity to DNA. The excellent properties of agarose from G. amansii of Taiwan confirm its potential diverse biotechnological applications. This innovative agarose preparation method with the significantly improved scale-efficiency can be modified for large-scale preparation of agarose for use in biotechnological industry and biochemical research.

제주도산 소라 (Batillus cornutus) 성패의 특정 해조류에 대한 선호도

This paper examines relation between growth of adult top shell and their selectivity over different marine algae. For the study the top shell density and the algal species in different depth were surveyed by SCUBA. To test selectivity over different types of algae, five algal species including Ecklonia cava, Undaria pinnatifida, Ulva spp, Gelidium amansii and Sargassum spp were provided to top shells in indoor water tank condition. In the field survey, the wet weight of Ecklonia cava was significantly (r = 0.612, p < 0.05) correlated to the shell length (SL) of top shell. In the indoor water tank experiment conducted over 12 hours after sunrise, adult top shell (SL > SL 4.5 cm) tended to select Ecklonia cava and Undaria pinnatifida, which are common and abundant algal in the subtidal waters in Jeju Island. Our field survey and the indoor experiment suggest that E. cava and U. pinnatifida could be food preferred by adult top shell in its habitat.

Macroalgal Community Structure on the Rocky Shores of Ongdo, Jusamdo, and Woejodo Islands of the Yellow Sea, Korea

The benthic algal community structures of the seaweed biomass, vertical distribution of dominant seaweeds, and species composition were examined on the rocky shores of Ongdo, Jusamdo, and Woejodo Islands, Korea, in August 2006. A total of 68 seaweeds were identified, comprising 5 green, 11 brown, and 52 red algae from the three study sites. The number of species at Ongdo (32 species) was less than that at Jusamdo (45 species) and Woejodo (44 species). Jusamdo exhibited the maximum seaweed biomass (73.99 g dry <TEX>$wt/m^2$</TEX>), while the minimum value was found at Woejodo (36.90 g dry <TEX>$wt/m^2$</TEX>). On the three islands, coarsely branched forms were the most dominant functional group in terms of species number and biomass among benthic algal species. The dominant species were Gelidium amansii, Chondrus ocellatus, and Chrysymenia wrightii at Ongdo, Sargasum thunbergii, Ulva pertusa, and Sargassum fusiformis at Jusamdo, and U. pertusa, Undaria pinnatifida, and Corallina pilulifera at Woejodo. Perennial seaweeds were abundant at Ongdo (G. amansii and C. ocellatus) and Jusamdo (S. thunbergii and S. fusiformis), whereas the sheet form of U. pertusa was relatively abundant at Woejodo Island.

한국 통영 연안의 계절적 해조상 및 군집구조 변화

Marine algal flora and community structure were examined at four sites at Tongyeong, Korea, from February to December 2009. Eighty-two algal species were identified, including 10 green, 25 brown, and 47 red algae. During the study period, the number of species was highest at the Dukpo (61 species) site followed by the Gajodo (60 species), Yepo (58 species), and Chilchundo (53 species) sites. Seaweed biomass ranged from 578.11 to 678.26 g, with the maximum and minimum at Chilchundo and Yepo, respectively. Dominant species in term of biomass were Sargassum thunbergii at Yepo, Dukpo, and Gajodo and Gelidium amansii at Chilchundo. The vertical distribution from the high to low intertidal zone was Sargassum thunbergii and Chondria crassicaulis; Sargassum thunbergii and Ulva linza; Ulva pertusa, Sargassum fulvellum, and Gelidium amansii. Two groups clearly segregated in a cluster analysis, a Yepo/Dukpo group (group A) and a Gajodo/Chilchundo (B), indicating relatively strong differences in similarity. The seaweed community structure of group A was characterized by higher biomass and higher species richness of brown algae than were observed in group B. Additionally, the biomass ratio and the species richness of green algae was lower in group A. These differences in seaweed community structure may have been resulted from the effects of cold effluents.

제주산 우뭇가사리(Gelidium amansii)로부터 DMSO 추출과 EDTA 수세법에 의한 상용화 아가로스 제조

Agar was prepared from Gelidium amansii collected from Jeju Island, South Korea. This agar preparation has high gel strength and low sulfate content compared with G. amansii agar from Morocco. Accordingly, agarose was made from the Jeju agar through the consecutive refining processes of dimethyl sulfoxide (DMSO) extraction and ethylene diamine tetra acetic acid (EDTA) washing. The physicochemical properties of the resulting agarose were compared with those from agarose prepared using only DMSO extraction. Consecutive DMSO extraction and EDTA washing more strongly affected the physicochemical properties of the agarose (purified agarose) compared with the use of DMSO extraction alone. These properties were similar to those of commercial agarose used for electrophoresis. In DNA electrophoresis, the separation and movement speed of the purified agarose were similar to those of the commercial agarose. In a <TEX>$^{13}C$</TEX> NMR analysis, the purified agarose exhibited the same carbon peak as the commercial agarose. When observed under scanning electron microscopy, the agar had an even and smooth surface without irregularities or pores, and the purified agarose had a wide surface area with a large number of pores; the commercial agarose had an irregular surface that would allow the solvent to easily permeate. These results illustrate that the physicochemical properties of agarose prepared from DMSO extraction and EDTA washing were more effective than those observed after DMSO extraction alone; thus, these processes used in succession will be useful in agarose industries.