Agar extraction process for Gracilaria cliftonii ()

Agar extraction process was developed for Gracilaria cliftonii by investigating the effects of various extraction variables and alkali treatments on agar yield and properties. The tested variables were soaking time, soaking temperature, seaweed to water ratio of, extraction temperatures and extraction time. Alkali treatments were carried out in alkali concentrations of 1%, 2%, 3% and 5% in a water bath at 60, 70 and 80 °C prior to agar extraction. The results showed that agar yield was significantly affected by all the tested variables. The agar yield was maximised when extraction process was carried out with 1 h soaking time at 30 °C with seaweed to water ratio of 1:150 and extracted for 3 h at 100 °C. The alkali-temperature combinations significantly influenced agar yield and properties. Irrespective of temperature, alkali treatments at 3% and 5% significantly increased the gel strength.     

Latitudinal variations of the yield and quality of agar from <Emphasis Type="Italic">Gelidium robustum</Emphasis> (Gelidiales,Rhodophyta) from the main commercial harvest beds along the western coast of the Baja California Peninsula,Mexico

The yield and quality of agar from Gelidium robustum from the main commercial harvest beds along the western coast of the Baja California Peninsula (Mexico) were evaluated within a latitudinal range of about 800 km (31°47′N to 27°05′N). Samples from six locations, Bahía Todos Santos, El Rosario, Isla de Cedros, Islas San Benito, Punta Eugenia, and Bahía Asunción, were analyzed. Bryozoan, protein, and agar content in the seaweed were estimated. The agar quality was determined by the content of 3,6-anhydrogalactose, sulfate, gel strength, and gelling and melting temperature. All the values of these variables were correlated and then with the satellite-derived data of the sea surface temperature (SST), net primary production (NPP), and photosynthetic active radiation (PAR) measured at each harvest bed during the summer months (June, July, and August) of 2000. In the northern and central region, the agar yield was 35% to 37% (Bahía Todos Santos, El Rosario, Isla de Cedros, and Islas San Benito), whereas lower yields were obtained from the southern beds (Punta Eugenia and Bahía Asunción). In contrast, the agar quality increased from the northern to the southern beds. A lower gel strength was obtained from Bahía Todos Santos and El Rosario (268 ± 16 and 205 ± 5 gcm⁻²) with a higher gel strength obtained from Isla de Cedros, Islas San Benito, and Bahía Asunción (384 to 444 ± 25 gcm⁻²). Yield was not correlated with the bryozoan content but was inversely correlated with the protein content in the seaweed. The sulfate content in the agar was inversely correlated with the gel strength and with the melting temperature. The 3,6-anhydrogalactose content showed slight variations among harvested beds. Analysis of satellite-derived data showed an equatorward increase of the SST, NPP, and PAR. The agar content correlated inversely with the equatorward increase of the NPP, whereas agar quality, i.e., gel strength, correlated positively with the NPP and PAR. No significant effects were observed on the yield and quality of agar with the latitudinal change of the SST.     

The seaweed hydrocolloid industry: 2016 updates,requirements, and outlook

The seaweed hydrocolloid industry, comprising agar, alginate, and carrageenan extracts, continues to grow in the order of 2–3% per year with the Asia-Pacific region increasingly dominating the raw material and manufacturing aspects of the industry. Geographic overviews, also in a historical perspective, of seaweed raw material availability including prices and consumption, manufacturing capacities, and utilizations and sales of extracts is presented. Some current and future industry dynamics, requirements, and changing structures, e.g., Indonesia’s increasingly dominant role within farming of agar and carrageenan-bearing seaweed species, randomly imposing of seaweed harvest restrictions or ban on exports, creation of a global certification standard for seaweed, and supply-demand dynamics for seaweed versus future global population are presented. The industry is increasingly being commoditized and China has become an important and, in many cases, dominant factor within all types of seaweed hydrocolloids and some explanations to this and strategic response by the rest of the industry is also touched upon. Also presented are some areas where the seaweed industry needs help from the scientific community. The main challenge is the ongoing general seaweed deterioration experienced in cultivated species—how are the strains to be improved and revitalized and can cultivation techniques be improved further? There is a general trend towards sustainability and, although seaweed cultivation and harvest can be sustainable, there is interest in the development of greener processes.     

Anti-tumorigenic components of a sea weed, Eeteromorpha clathrata

Anti-tumorigenic action of compounds in some seaweed was evaluated by using selective cytotoxicity against a transformed mouse 3T3 cell (SV-T2) and its normal counter part. The extracts of seaweeds (Wakame, Konbu, Mirin, Aosa, and Sujiaonori) were added to the cells in 96 well microplate. The viability of the cell was measured at 24 hr after sample addition. Of the five weeds, the ethanol extracts from Sujiaonori exhibited the selective cytotoxicity. The ethanol extracts from Sujiaonori was applied to gel permeation chromatography on Sephadex G-50. The anti-cancer activity was detected in two fractions with high molecular weight near void volume and with small molecular weight. The high molecular weight fractions had absorbance at 280 nm, suggesting that they contained proteineous substances. The small molecular weight fractions were estimated to be around 500 Dalton.     

Agar from the red seaweed,Laurencia flexilis (Ceramiales,Rhodophyta) from northern Philippines

The worldwide production of the gelling agent agar mainly rely on the red algae of the order Gracilariales and Gelidiales for raw material. We investigate here the potential of a species from another red algal order, Ceramiales as an agar source. The agar from Laurencia flexilis collected in northern Philippines was extracted using native and alkali treatment procedures and the properties of the extracts were determined using chemical, spectroscopic and physical methods. The native agar, 26% dry weight basis, forms a gel with moderate gel strength (200 g cm^?2). Alkali‐treatment did not enhance the gel strength, indicating insignificant amounts of galactose‐6‐sulfate residue, the precursor of the gel‐forming 3,6‐anhydrogalactose (3,6‐AG) moieties. Furthermore, the Fourier transform infrared and chemical analysis showed low sulfate and high 3,6‐AG levels, not affected significantly by the alkali treatment. Nuclear magnetic resonance spectroscopic analysis revealed 3‐linked 6‐O‐methyl‐D‐galactose and 4‐linked 3,6‐anhydro‐L‐galactose as the major repeating unit of the native extract, with minor sulfation at 4‐position of the 3‐linked galactose residues. The native and alkali treated agars have comparably high gelling and melting temperatures, whereas the former exhibits higher gel syneresis. Laurencia flexilis could be a good source of agar that possesses physico‐chemical and rheological qualities appropriate for food applications. Due to the inability of alkali treatment to enhance the key gel qualities of the native extract, it is recommended that commercial agar extraction from this seaweed would be done without pursuing this widely‐used industrial procedure.     

Brown seaweed protein as an inhibitor of marine mollusk endo-(1-->3)-beta-D-glucanases

Aqueous ethanol extracts from brown seaweed were found to contain substances inhibiting endo-(1-->3)-beta-D-glucanases, the digestive enzymes of marine mollusks. The inhibitors were detected in 70% of the brown seaweeds investigated. An irreversible protein inhibitor with high specificity for endo-(1-->3)-beta-D-glucanases of marine mollusks was isolated from the brown seaweed, Laminaria cichorioides. As determined by gel filtration, the molecular weight of the inhibitor was 46 kDa. The value of [I]50 (10(-8) M) for the inhibitor was comparable with the corresponding value for natural alpha-amylase inhibitors from terrestrial plants. Chemical modification results indicated that tryptophan, dicarboxylic acid, histidine and probably tyrosine residues of inhibitor molecule are important for interaction of the inhibitor with the enzyme.     

Chemical and physical characteristics of carrageenan extracted from Eucheuma spinosum harvested from three different Indonesian coastal sea regions

Carrageenan extracted from Eucheuma spinosum harvested from three different coastal sea regions, where this alga has been mainly cultivated, were determined for their chemical and physical characteristics. The carrageenan was extracted from the seaweed using hot alkali followed by precipitation, drying, and milling. The carrageenan properties were determined in terms of yield, ash, mineral, sulfate content, functional group, molecular weight, and viscosity profile. Physical characteristics of carrageenan were evaluated by a texture analyzer for gel strength and a rapid visco analyzer for viscosity. The yield of carrageenan from Sumenep (34.81 ± 5.83%) and Takalar (37.16 ± 3.26%) was found to be relatively higher than that of Nusa Penida (25.81 ± 1.93%). The calcium content was higher than magnesium, potassium and sodium content, and no cadmium, lead, mercury, and arsenic detected in all carrageenan. The ash content was around 29%; while, the sulfate content was in the range of 30–32%, and those were not different in all carrageenan. The presence of sulfate content was identified by FTIR at absorption band of 1373 cm^?1. It was found that the molecular weight of carrageenan from Takalar were relatively higher and the gel strength of carrageenan from Takalar were significantly higher than that of carrageenan from Nusa Penida and Sumenep. Likewise, upon cooling from 80 to 20°C, the viscosity profile of carrageenan from Takalar characterized by higher viscosity compared to that of carrageenan from Sumenep and Nusa Penida. These results indicated that carrageenan from Nusa Penida, Sumenep, and Takalar were identified as iota‐carrageenan with similar physico‐chemical characteristics except for the gel strength, viscosity profile upon cooling from 80 to 20°C and the yield.     

Purification and properties of α-1,4-glucan phosphorylase from the red seaweed Gracilaria sordida (Gracilariales)

α-1,4-Glucan phosphorylase (EC 2.4.1.1) from the red seaweed Gracilaria sordida (Harv.) W. Nelson was adsorbed onto starch-Sepharose 6B and Sephacryl S-300 under specified conditions. The algal enzyme was purified to homogeneity by these two steps. A molecular weight of 97.4 kDa was observed on SDS-polyacrylamide gel electrophoresis under reducing conditions, while the native molecular weight was 240 kDa asrevealed by 8-25% native gradient gel electrophoresis or 245 kDa by gel filtration. The pI of the enzyme was 5.4. It had a Km of 227, 264, 285, and 453 μg ml-1, respectively, towards glycogen, amylopectin, amylose, and maltodextrin. The enzyme activity was inhibited by cyclohexaamylose, ADP-glucose, and UDP-glucose. In contrast to other plant sources, cell-free extracts of G. sordida contained only one form of phosphorylase.     

Effect of dark treatment on the starch degradation and the agar quality of cultivated Gracilariopsis lemaneiformis (Rhodophyta,Gracilariales) from Venezuela

The effect of darkness and ammonia on Gracilariopsis lemaneiformis agar yield and quality was studied. Plants were cultivated under controlled conditions in the laboratory and fertilized with ammonia prior to a month storage in the dark. Starch inclusions are known to interfere with the mechanical properties of the isolated agar. The starch content of algal tissue and the activities of the floridean starch phosphorylase and -glucosidase were measured during this storage period as a way to follow the enhancement of starch degradation in the absence of light, and to measure the effect of darkness on the content and quality of the synthesized agar. The agar yield, gel strength, 3,6-anhydrogalactose, sulphate and starch content were considerably affected by this dark treatment. The values obtained reflect a pattern of optimization of agar quality when Gracilariopsis lemaneiformis was stored in the dark. The isolated agar following dark treatment is less contaminated by starch inclusions. This is shown by the improvement in the agar yield and quality, the nutrient status of the plant and the reduction of the starch content.Abbreviations DW: Dry weight - FW: Fresh weight - S: Sulphate - 3,6-AG: 3,6-anhydrogalactose - GS: Gel strenght at 1.5% concentration - CX: Cold extract at 25 °C - A: Autoclaved extract at 121 °C - MW: Molecular weight - PFD: Photon flux density     

重组hepcidin的分离纯化及鉴定

建立了重组hepcidin的分离纯化方法,并鉴定其抗菌活性。经金属螯合初步纯化的重组蛋白在cysteine/cystine氧化还原体系中氧化形成二硫键,用变性条件下的凝胶过滤除去多聚体,稀释复性后用于肠激酶酶切反应,得到重组hepcidin。融合蛋白His-hepcidin经氧化、复性后的总收率为50%,纯度大于95%。酶切后所得重组hepcidin经抑菌圈试验检验,对枯草芽孢杆菌具有抗菌活性。LC-ESI-MS与园二色光谱检测显示重组hepcidin与天然hepcidin相对分子质量相同、二级结构相似。     

Studies on thyroid hormone-binding proteins. I. The subunit structure of human thyroxine-binding globulin and its interaction with ligands.

Thyroxine-binding globulin was isolated from human plasma by ammonium sulfate fractionation, chromatographic separations on diethylaminoethyl-Sephadex, gel chromatography, and two different electrophoretic procedures. The highly purified was homogeneous when subjected to polyacrylamide gel electrophoresis, ultracentrifugation analyses, and immunochemical determinations. The weight average molecular weight as determined by sedimentation equilibrium ultracentrifugations was 54,000 and by sedimentation diffusion data 55,000. Amino acid analyses indicated a minimum of 110 amino acid residues per molecule. By determination of the minimum in the curve for the fraction of maximum deviation from the amino acid analyses it was found that the minimum molecular weight for the polypeptide was 12,200. Carbohydrate analyses demonstrated the presence f equimolar amounts of amnnose, galactose, and glucosamine, and the carbohydrate portion constituted 7.5% of the total weight. The amino acid analyses suggested that thyroxine-binding globulin is composed of 4 subunits. Molecular weight determinations by gel chromatography in 6 M guanidine hydrochloride indicated the presence of three species of globulin with apparent molecular weights 52,000, 25,000, and 13,500, respectively. Prolonged storage in guanidine hydrochloride promoted a more than 60% yield of the monomeric species. Moreover, a half-molecule of thyroxine-binding globulin was isolated and shown to consist of two polypeptide chains of similar molecular weight...     

Development of an eco-friendly agar extraction technique from the red seaweed Gracilaria lemaneiformis

The red seaweed, Gracilaria lemaneiformis growing as an aquaculture bioremediator along the coasts of Liaodong Peninsula, China, was investigated for the agar production. An eco-friendly method called agar photobleaching extraction process was developed for the benefit of workers' health and safety of the environment. The native agar (NA), alkali-modified agar (AA), chemical-bleached agar (CA) and photobleached agar (PA), which were extracted using different processes, were evaluated for their physical and chemical properties. The PA showed most desirable performances in terms of gel strength, gelling temperature, sulfate content and 3,6-anhydro-l-galactose content. Among the different processed agars, PA gel strength was 1913 g/cm2, the highest among the different processed agars, which increased 8.6% on the basis of the AA. Further we applied this new technique to extract agars from Gracilaria asiatica, and similar results were obtained with that of G. lemaneiformis. This indicates that the agar photobleaching extraction process is a feasible method for Gracilaria species and has a potential application. During the whole agar photobleaching extraction process the pigment content of G. lemaneiformis declined gradually and the TOC concentration in photobleaching solution increased along with the increase in the irradiation time. The mechanism of agar photobleaching could be elucidated by the photolysis theory.     

Agar Processed from Gracilaria foliifera of Florida

Agar producing a firm gel with specific rheological properties as well as a good commercial value was processed from Gracilaria foliifera (Forsskal) Børgesen, a red alga harvested along the Florida west coast. The alkali pretreatment of the seaweed based on the Funaki-Kojima’s method was applied before the process of agar extraction. The chemical and physical properties of the processed agar were compared with other typical agar experimentally processed.     

Purification and characterization of a molybdenum-pterin-binding protein (Mop) in Clostridium pasteurianum W5.

A large-scale fractionation scheme purified the major molybdenum(Mo)-binding protein (Mop) from crude extracts of Clostridium pasteurianum, with a 10 and 0.2% yield of Mo and protein, respectively. The apparent molecular weight of the purified molybdoprotein is 5,700, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein contains 0.7 mol of Mo per mol of protein with a molecular weight of 5,700. Mop, as isolated, has a peak absorbency at 293 nm. Denaturation and oxidation of the molybdoprotein released multiple pterin like fluorescent compounds. Mop appears to contain a pterin derivative and Mo, but phosphate analysis indicated that the pterin at the very least is not phosphorylated; phosphorylation is required for functional molybdenum cofactor. All treatments used to release the putative Mo-pterin species from Mop failed to yield a molybdopterin that had detectable molybdenum cofactor activity.     

Purification of undegraded ceruloplasmin from outdated human plasma

A method for the rapid isolation of homogeneous undegraded ceruloplasmin from outdated human plasma is reported. The procedure consists of a precipitation step with polyethylene glycol 4000, batchwise adsorption and elution from QAE-Sephadex, and gradient elution from DEAE-Sepharose CL-6B. Ceruloplasmin was purified 1740-fold and the yield from outdated plasma was 67%. The purified ceruloplasmin was found to be homogeneous on anionic polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulfate-PAGE, isoelectric focusing, and low-speed equilibrium centrifugation. The isoelectric point as determined by isoelectric focusing was 4.4. The purified enzyme was sensitive to storage; when a sample was resubmitted to PAGE after 4 months of storage at 4 degrees C, two bands were obtained and the fast-moving band showed no oxidase activity. The molecular weight estimated by gel electrophoresis and sedimentation equilibrium centrifugation was 130,000.     

Purification, characterization, and immunological properties of fumarase from Euglena gracilis var. bacillaris.

A rapid three-step procedure utilizing heat treatment, ammonium sulfate fractionation, and affinity chromatography on Matrex gel Orange A purified fumarase (EC 4.2.1.2) 632-fold with an 18% yield from crude extracts of Euglena gracilis var. bacillaris. The apparent molecular weight of the native enzyme was 120,000 as determined by gel filtration on Sephacryl S-300. The preparation was over 95% pure, and the subunit molecular weight was 60,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that the enzyme is a dimer composed of two identical subunits. The pH optimum for E. gracilis fumarase was 8.4. The Km values for malate and fumarate were 1.4 and 0.031 mM, respectively. Preparative two-dimensional gel electrophoresis was used to further purify the enzyme for antibody production. On Ouchterlony double-immunodiffusion gels, the antifumarase serum gave a sharp precipitin line against total E. gracilis protein and purified E. gracilis fumarase. It did not cross-react with purified pig heart fumarase. On immunoblots of purified E. gracilis fumarase and crude cell extracts of E. gracilis, the antibody recognized a single polypeptide with a molecular weight of approximately 60,000, indicating that the antibody is monospecific. This polypeptide was found in E. gracilis mitochondria. The antibody cross-reacted with an Escherichia coli protein whose molecular weight was approximately 60,000, the reported molecular weight of the fumA gene product of E. coli, but it failed to cross-react with proteins found in crude mouse cell extracts, Bacillus subtilis extracts, or purified pig heart fumarase.     

Stabilization and purification of ornithine transcarbamylase from Neisseria gonorrhoeae.

Ornithine transcarbamylase was stabilized in cell-free extracts by the presence of either carbamyl phosphate or glycerol. The enzyme was rapidly purified by a procedure consisting of ion-exchange chromatography and electrofocusing. The native molecular weight of the enzyme was determined by gel filtration to be 110,000. A subunit molecular weight of 36,000 was determined by polyacrylamide electrophoresis under dissociating conditions. These findings indicated a trimeric quaternary structure for the enzyme. The isoelectric point of the purified enzyme was 4.75, and no evidence of multiple forms of active enzyme was found in either crude or purified preparations. An inactive form of the enzyme appeared upon storage in the absence of stabilization buffer.     

Does storage time influence yield and agar properties in the tropical agarophyte Gracilaria cornea?

The agar yield and quality characteristics of Gracilaria cornea from Yucatán, Mexico, werestudied during 18 months of storage. Biomass wasstored at a temperature of 22.1 ± 0.9 ^°Cand humidity of 59.8±3.6%. The agar contentvaried erratically, but the average value waspractically constant over the storage period with an average of 20.1 ± 1.5±. Gel strength, gelling and meltingtemperatures were negatively affected by the totalstorage time. No significant changes were found duringthe first five months for these characteristics withmean values of 1134 ± 57 g cm^-2, 40.8 ±0.4 ^°C and 91.2 ± 0.9 ^°Crespectively. Agar degradation was evident after thefifth month and accounted for a 17± loss in gelstrength and 7± in gelling and meltingtemperatures. Nevertheless, gel strength valuesremained around 930 ± 23 g cm^-2 with nosignificant changes until the end of the storageperiod. The decrease in gel strength showed asignificant relationship with decrease in3,6-anhydrogalactose but not variation in sulphatecontent. This was probably due to agar hydrolysiscaused by enzymatic processes of endogenous and/ormicrobial origin. These results suggest that thetropical G. cornea had a similar resistance todegradation during storage to that observed for G. chilensis, a cold water species. Agar qualityand yield in G. cornea after one and a half yearof storage are within the range of food grade agar.     

Purification and properties of an NADP-specific 6-phosphogluconate dehydrogenase from Streptococcus faecalis.

A procedure is described for the purification of 6-phosphogluconate dehydrogenase (6-phospho-D-gluconate:NADP oxidoreductase (decarboxylating) EC 1.1.1.44) from cell extracts of Streptococcus gaecalis. A 180-fold purification was achieved with an over-all yield of about 12% and an average specific activity of 14. The enzyme was homogeneous as determined by polyacrylamide gel electrophoresis, immunoelectrophoresis, and sedimentation equilibrium, studies. Its weight average molecular weight, as measured by sedimentation equilibrium, was 108,000 +/- 3,600. Other methods employed for molecular weight determinations gave values that ranged between 106,000 and 115,000. An analysis of the enzyme by sodium dodecyl sulfate polyacrylamide gel electrophoresis showed it to be a dimer composed of subunits having equal molecular weight. The amino acid composition of the streptococcal enzyme is reported. The apparent Km values for NADP and 6-phosphogluconate were calculated from kinetic data and found to be 0.015 mM and 0.024 mM, respectively. Kinetic studies also indicated that the binding of one substrate did not affect the apparent affinity of the enzyme for the other substrate.     

On the properties of agar gel containing ionic and non-ionic surfactants

Rheological and thermal properties of agar sol and gel in presence of various cationic, anionic and non-ionic surfactants are reported. The agar used was from the red seaweed Gelidiella acerosa. The gel strength, viscosity, rigidity (G'), gelling temperature and melting temperature were observed to decrease in presence of non-ionic surfactants whereas these were enhanced in presence of ionic surfactants. TGA studies showed that 1.5% agar gels containing non-ionic surfactants lose water at a lower temperature than the control agar gel whereas gels containing ionic surfactants hold on to water more tenaciously. DSC studies, on the other hand, show that the gel to sol transition occurs at lower temperatures in presence of non-ionic surfactants and at higher temperature in presence of ionic surfactants when compared with the control gel. The non-ionic surfactants, Triton X-100 and Brij 35, enabled relatively concentrated agar extractive to be filtered readily, as a result of which water usage in the process could be reduced by 50%. The surfactant was subsequently removed through freeze-thaw operations to restore the gelling capacity of the agar. The finding that 0.3-0.4% (w/v) sodium lauryl sulfate (SLS) lowers the sol-gel transition temperature from 41 to 36 degrees C without adversely affecting gel strength is another useful outcome of the study that may enable better formulations of bacteriological agar to be prepared.