Improved isolation of glucuronan from algae and the production of glucuronic acid oligosaccharides using a glucuronan lyase

A new source for the production of bioactive glucuronic acid oligosaccharides (GlcUAOs) from the depolymerization of green seaweed Ulva lactuca glucuronan (Algal glucuronan) has been investigated. Algal glucuronan purification was optimized by the acidic precipitation method which allowed us to separate the polysaccharide mixture extracted from the cell wall of Ulva lactuca using hot water containing sodium oxalate. A series of the GlcUAOs were obtained by enzyme degradation of algal glucuronan with a glucuronan lyase (GL) isolated from Trichoderma strain. The putative bioactive GlcUAOs generated were then purified by size-exclusion chromatography in gram quantity and characterized by ¹H/¹³C NMR spectroscopy and ESI-Q/TOF-mass spectrometry.     

New species of freshwater Ulva, Ulva limnetica (Ulvales, Ulvophyceae) from the Ryukyu Islands, Japan

Ulva limnetica Ichihara et Shimada, sp. nov. (Ulvales, Ulvophyceae) is described from the Ryukyu Islands, Japan, and is characterized by thalli that are: (i) branched, tubular, fragile and wrinkled; (ii) up to 80 cm in height and up to 2 cm in diameter; (iii) light to yellowish green in color; and (iv) having an asexual reproduction by means of quadriflagellate swarmers. Rhizoidal cells bear tubular extensions on the outside of the cell layer in the stipe. Ulva limnetica is distinguished from species with similar thalli by chloroplast disposition, branching pattern, number of pyrenoids per cell and gross morphology. It is also distinguished by sequences of the nuclear-encoded 18S ribosomal RNA gene, internal transcribed spacer 2 region and the plastid-encoded large subunit of ribulose-1,5-bisphosphate carboxylase/oxgenase gene ( rbc L). Ulva limnetica was clustered with other Ulva species in an early diverging lineage within the genus.     

THE HORMESIS OF THE GREEN MACROALGA ULVA FASCIATA WITH LOW‐DOSE 60COBALT GAMMA RADIATION1

Homogenous germlings of the marine macroalga Ulva fasciata D. (synonym, Ulva lactuca L.) were used to study hormesis effects in macroalgae grown under a low dose of ⁶⁰Co γ‐ray radiation. The results of this study are the first to confirm the effects of macroalgal hormesis. Here it was demonstrated that growth of U. fasciata germlings was promoted substantially under 15 Gy of ⁶⁰Co γ‐ray radiation, with an average increase of algal biomass of 47.43%. The levels of polysaccharides and lipids varied among the tested material and showed no effects from the ⁶⁰Co γ‐ray radiation. However, the amount of protein was higher in the irradiated algae than in the control; the highest protein content of the irradiated algae was 3.958% (dry weight), in contrast to 2.318% in nonirradiated samples. This technique was applied to a field algal mass culture, which decreased the harvest time from 90 to 60 d. The mass culture approach may facilitate the production of macroalgae under unstable weather conditions such as typhoons in the summer or strong waves in the winter. The mass‐cultured macroalgae could be used as a source of bioenergy through the fermentation of algal simple sugars that derived from polysaccharides to produce ethanol.     

TRUE IDENTITY OF THE EUROPEAN FRESHWATER ULVA (CHLOROPHYTA,ULVOPHYCEAE) REVEALED BY A COMBINED MOLECULAR AND MORPHOLOGICAL APPROACH1

A set of 18 freshwater and morphologically similar marine samples of Ulva were collected from inland and coastal waters throughout Europe to assess their taxonomic identity and invasive potential. An additional 11 specimens were obtained from herbaria. The material was studied using a combination of classical morphological methods and molecular techniques; the latter included sequencing of the nuclear internal transcribed spacer (ITS) region (ITS1‐5.8S‐ITS2) and the chloroplast RUBISCO LSU (rbcL) gene and comparison of the ITS2 secondary structure predictions. Based on classical methods, all the specimens could be determined as U. flexuosa Wulfen and could be further divided into three groups matching three infraspecific taxa. This pattern was generally well supported by molecular phylogenetic analyses. All sequenced samples formed a monophyletic lineage within Ulva, showing a putative synapomorphy in the ITS2 secondary structure. The individual subspecies corresponded to phylogenetic clusters within this lineage. In freshwater habitats, the dominant taxon was U. flexuosa subsp. pilifera, but subsp. paradoxa was also occasionally recorded. In marine habitats, only U. flexuosa subsp. flexuosa and subsp. paradoxa were located. These findings support the view that U. flexuosa subsp. pilifera is primarily a freshwater alga that probably dominates in Europe. As confirmed by the study of herbarium specimens, U. flexuosa should be regarded as indigenous, although it has a tendency to form blooms under certain conditions. Besides clarifying the identity of prevailing European freshwater Ulva, the study provides novel data concerning the distribution and morphological plasticity within the U. flexuosa complex.     

Ulva flexuosa (Ulvaceae,Chlorophyta) inhabiting inland aquatic ecosystems: molecular,morphological and ecological discrimination of subspecies

Species of Ulva (Ulvaceae, Chlorophyta) are relatively common macroalgae in most coastal areas. In this paper, we examine Ulva in inland aquatic ecosystems to determine relationships between the chemical parameters of the water and the Ulva taxa present. Two groups of samples of freshwater Ulva were selected from: (i) habitats with high salinity levels, and (ii) habitats with fresh or slightly saline waters. We showed that these two groups of Ulva, which occupy distinct habitats, represent two separate taxa. The first group represents Ulva flexuosa subsp. pilifera (Kützing) M.J. Wynne, which is very common and regularly encountered in fresh waters, but is equally well represented in slightly saline waters. The second is U. flexuosa subsp. paradoxa (C. Agardh) M.J. Wynne, which is less common, preferring mainly salt-wedge saline waters. We did not find both Ulva flexuosa subspecies together at any of the research sites.

There were significant morphological differences between the Ulva flexuosa subspecies. Ulva flexuosa subsp. pilifera had longer and wider thalli and wider cells than those of subsp. paradoxa. Ulva flexuosa subsp. paradoxa cells were longer and of greater circumference than those of subsp. pilifera. Analyses of the rRNA ITS region and partial rbcL gene sequences demonstrated minor sequence variation within the two subspecies. The results of this study provide detailed information on the characteristics of limnic Ulva habitats and indicate the niches where such populations are likely to occur.     

Application of response surface method to evaluate the cytotoxic potency of Ulva fasciata Delile,a marine macro alga

Bioprospecting of marine natural products has recently produced a substantial number of drug candidates. Ulva fasciata Delile, belonging to the family Ulvaceae, is a green marine macro alga that grows profusely on the coastal seashore of South India. In the present study, we investigated the in vitro cytotoxic potential of a methanolic extract of U.fasciata Delile (MEUF) using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay against human colon carcinoma (HT-29), human hepatocyte carcinoma (Hep-G2), and human breast carcinoma (MCF-7) cell lines. Response surface methodology (RSM) was applied using central-composite experimental design (CCD) to obtain optimum combined effect of concentration and cancer cells with highest cytotoxicity. The effect of concentration, cancer cell lines as independent variables on absorbance (OD), percent cell survival and percent cell inhibition as dependent variables was investigated. Maximum cytotoxic activity of MEUF was established for Hep-G2 with lowest OD or percent cell survival; highest percent cell inhibition with significant difference (p > 0.05) was compared to HT-29 and MCF-7.     

Production of recombinant protein in Escherichia coli cultured in extract from waste product alga,Ulva lactuca

This study examined the potential for waste product alga, Ulva lactuca, to serve as a media component for recombinant protein production in Escherichia coli. To facilitate this investigation, U. lactuca harvested from Jamaica Bay was dried, and nutrients acid extracted for use as a growth media. The E. coli cell line BL21(DE3) was used to assess the effects on growth and production of recombinant green fluorescent protein (GFP). This study showed that media composed of acid extracts without further nutrient addition maintained E. coli growth and recombinant protein production. Extracts made from dried algae lots less than six‐months‐old were able to produce two‐fold more GFP protein than traditional Lysogeny Broth media. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:784–789, 2014     

盐度、光照和营养盐对孔石莼(Ulva pertusa)光合作用的影响

通过测定光合作用产氧速率研究了孔石莼Ulvapertusa不育性变种在不同盐度、光强度和营养盐水平下的光合作用特性.结果表明,盐度可影响光合作用速率,在2710lx光强下孔石莼在盐度20‰左右有最大光合作用速率;光合作用参数Pm为79.62O2,μg/cm2*h,Is为191.08μE/m2*s,IC为10.12μE/m2*s;对营养盐(NH4+-N,NO2--N,PO43--P)的吸收特征可用Michaelis-Menten方程描述,低光强(106lx)下氨氮超过0.07mmol/L时可抑制孔石莼的光合作用.     

UV-B辐射增强对海洋大型藻与微型藻种群生长关系的影响

选用孔石莼和青岛大扁藻为海洋大型藻和微型藻的代表,通过室内添加模拟试验研究了UV-B辐射增强对孔石莼(重量固定)与青岛大扁藻(密度不同)种群生长关系的影响。结果表明:(1)在单养情况下,4个UV-B辐射剂量都对孔石莼的生长产生抑制作用;对青岛大扁藻生长的影响却不同,低剂量(U-1)的UV-B辐射对青岛大扁藻的生长有促进作用,而高剂量的UV-B辐射则有显著的抑制作用;且因初始接种密度不同而各异。(2)在共养情况下,微藻对孔石莼的生长表现出一定的抑制作用,随着微藻初始接种密度的增加,其抑制作用亦增加;反之,在共培养的初始阶段(6 d内)孔石莼对微藻的生长也有抑制作用,但后期阶段(9 d后)表现出促进作用。(3)在共培养的同时附加UV-B辐射处理,随着初始接种密度的增加,青岛大扁藻对孔石莼生长的抑制作用更加明显;同时,与共养相比较,孔石莼对微藻生长的抑制作用亦趋于明显。     

Allelopathic effects of Ulva lactuca on selected species of harmful bloom-forming microalgae in laboratory cultures

Allelopathic effects of the green macroalgae Ulva lactuca on the growth of three species of red tide microalgae, Heterosigma akashiwo, Alexandrium tamarense, and Skeletonema costatum were tested in laboratory co-cultures precluding the nutrient and light limitation and the effect of high pH. The growth of all three species of microalgae was significantly (p < 0.01) inhibited by fresh U. lactuca. In nutrient replete semicontinuous co-cultures with U. lactuca, H. akashiwo was completely dead in 12 days, and the growth of A. tamarense and S. costatum was reduced by 48 and 46%, respectively by U. lactuca within 12 days. The U. lactuca culture filtrate exhibited a significant (p < 0.05) inhibitory effect on the microalgae in the first 1 or 2 days, but growth resumed in the following days, and S. costatum growth was slightly (p > 0.05) promoted from day 3. The results suggested that the allelopathic compounds are quickly degradable and a long-term inhibition might need the continuous addition of compounds originated from macroalgae. Dried U. lactuca also exhibited inhibitory effects on the microalgae, and the normalized mean growth rates of microalgae decreased with the biomass of dried U. lactuca. The dependent relationships were y = −2.1208x² + 1.0159x + 0.9752 for H. akashiwo, y = 0.7133x² − 3.5813x + 1.1665 for A. tamarense, and y = −0.2114x² − 1.063x + 1.0873 for S. costatum, respectively. The potential feasibility of utilization of dried U. lactuca against red tide microalgae was 2.0 g dry wt L⁻¹. The present study shows that U. lactuca exhibits negative allelopathic effects on harmful bloom-forming microalgae.