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Structural analysis and anti-complement activity of polysaccharides extracted from Grateloupia livida (Harv.) Yamada

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Journal of Oceanology and Limnology 2019年第3期37卷 806-814,I0001-I0002页

作者： ZHANG Wenjing JIN Weihua duan delin ZHANG Quanbin Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China School of Medicine Zhejiang UniversityHangzhou 310058China College of Biotechnology and Bioengineering Zhejiang University of TechnologyHangzhou 310014China Laboratory for Marine Biology and Biotechnology Qingdao National Laboratory for Marine Science and TechnologyQingdao 266071China State Key Lab of Seaweed Bioactive Substances Qingdao 266000China Center for Ocean Mega-Science Chinese Academy of SciencesQingdao 266071China

Polysaccharides were extracted from Grateloupia livida(Harv.) Yamada using hot water(extracted product denoted WGW) and then degraded in dilute sulfuric acid(degraded product denoted WGWD). The degraded mixture was then separated into four fractions through anion exchange chromatography on 2-diethylaminoethanol(DEAE)-Bio-Gel Agarose FF gel. Electrospray ionization collision-induced dissociation tandem mass spectrometry(ESI-CID-MS/MS) was performed to elucidate the structural features of all fractions. In combination with nuclear magnetic resonance spectroscopy(NMR)and infrared spectroscopy(IR) data, the major polysaccharide structures were concluded to be μ-carrageenan and κ-carrageenan. μ-Carrageenan usually has a backbone of alternating 1,3-linked β-D-galactopyranose residues sulfated at C-4 and 1,4-linked a-D-galactopyranose residues sulfated at C-6, while κ-carrageenan consists of alternating 1,3-linked β-D-galactopyranose residues sulfated at C-4 and 1,4-linked a-D-3,6-anhydrogalactopyranose residues. Trace v-carrageenan, composed of 1,3-linked β-D-galactopyranose residues sulfated at C-4 and 1,4-linked a-D-galactopyranose residues sulfated at C-2 and C-6, was also detected. Furthermore, the polysaccharide had a backbone comprising 1,3-linked β-D-galactopyranose and1,4-linked α-L-galactopyranose sulfated at C-6, which is the agarose precursor. The hydroxy groups in the galactopyranose were partially substituted by methyl and pyruvic acid acetal(PA) groups. The anticomplementary activities of WGW and its derivatives against classical pathways were measured. The native polysaccharides in WGW had higher activities, while the derivatives had much weaker activities. This indicated that the molecular weight and sulfate content were important factors affecting the anti-complement activity.

关键词： red alga Grateloupia livida(Harv.)Yamada mass spectrometry structure

Preparation and characterization of agar, agarose,and agaropectin from the red alga Ahnfeltia plicata

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Journal of Oceanology and Limnology 2019年第3期37卷 815-824页

作者： ZHANG Yu FU Xiaoting duan delin XU Jiachao GAO Xin College of Food Science and Engineering Ocean University of ChinaQingdao 266003China Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China State Key Lab of Seaweed Bioactive Substances Qingdao 266400China

Agar,agarose,and agaropectin were extracted from the red alga Ahnfeltia plicata,and their properties and structures were *** was extracted by a comparatively low alkaline consumption of 1.2%.It exhibited a gel strength of 1 152.50±74.25 g/cm^2 and a sulfate content of 0.55%±0.08%.The yield of agar from *** was 24.53%,which is higher than those of other agarophytes commonly used in *** kinds of the method were compared for the purification of agarose,and the physicochemical properties of agarose that was prepared under the optimal condition were identical to those of commercially available ***,agaropectin was purified from *** and characterized by GC,HPLC,UV-spectrum,and FI-IR to understand its composition and *** was the first time to comprehensively study the agar and its fractions from the red alga of *** research provided an eco-friendly agar extraction method from *** and revealed its potential application for the production of agar,agarose,and agaropectin.

关键词： Ahnfeltia plicata agar agarose agaropectin

Isolation, Expression and Characterization of rbc L Gene from Ulva prolifera J. Agardh(Ulvophyceae, Chlorophyta)

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Journal of Ocean University of China 2015年第6期14卷 1087-1095页

作者： SHAO Zhanru LI Wei GUO Hui duan delin Institute of Oceanology Chinese Academy of Sciences Institute of General Botany and Plant Physiology Friedrich Schiller University University of Chinese Academy of Sciences

Ulva prolifera is a typical green alga in subtidal areas and can grow tremendously fast. A highly efficient Rubisco enzyme which is encoded by Up Rbc L gene may contribute to the rapid growth. In this study, the full-length Up Rbc L open reading frame(ORF) was identified, which encoded a protein of 474 amino acids. Phylogenetic analysis of Up Rbc L sequences revealed that Chlorophyta had a closer genetic relationship with higher plants than with Rhodophyta and Phaeophyta. The two distinct residues(aa11 and aa91) were presumed to be unique for Rubisco catalytic activity. The predicted three-dimensional structure showed that one α/β-barrel existed in the C-terminal region, and the sites for Mg^(2+)coordination and CO_2 fixation were also located in this region. Gene expression profile indicated that Up Rbc L was expressed at a higher level under light exposure than in darkness. When the culture temperature reached 35℃, the expression level of Up Rbc L was 2.5-fold lower than at 15℃, and the carboxylase activity exhibited 13.8-fold decrease. Up Rbc L was heterologously expressed in E. coli and was purified by Ni^(2+) affinity chromatography. The physiological and biochemical characterization of recombinant Rubisco will be explored in the future.

关键词： Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit sequence analysis real-time PCR in vitro expres-sion Ulva prolifera

Genetic Mapping of Laminaria japonica and L. Iongissima Using Amplified Fragment Length Polymorphism Markers in a ＂Two-Way Pseudo- Testcross＂ Strategy

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Journal of Integrative Plant Biology 2007年第3期49卷 392-400页

作者： Yuhui Li Yingxia Yang Jidong Liu Xiuliang Wang Tianxiang Gao delin duan Ocean University of China Qingdao 266003 China Institute of Oceanology the Chinese Academy of Sciences Qingdao 266071 China Ludong University Yantai 264025 China

With a ＂two-way pseudo-testcross＂ mapping strategy, we applied the amplified fragment length polymorphism （AFLP） markers to construct two moderate density genetic linkage maps for Laminaria. The linkage maps were generated from the 60 progenies of the F1 cross family （Laminaria iongissima Aresch. × L. Japonica Miyabe） with twenty pairs of primer combinations. Of the 333 polymorphic loci scored in 60 progenies, 173 segregated in a 1：1 ratio, corresponding to DNA polymorphisms heterozygous in a single parent, and the other 58 loci existing in both parents followed a 3：1 Mendelian segregation ratio. Among the loci with 1：1 segregating ratios, 79 loci were ordered in 14 linkage groups （648.6 cM） of the paternal map, and 72 loci were ordered in 14 linkage groups （601.9 cM） of the maternal map. The average density of loci was approximately 1 per 8 cM. To Investigate the homologies between two parental maps, we used 58 loci segregated 3：1 for further analysis, and deduced one homologous linkage group. The linkage data developed in these maps will be useful for detecting loci-controlling commercially important traits for Laminaria.

关键词： amplified fragment length polymorphism makers genetic linkage map Laminaria japonica Laminaria longissima segregation ratio two-way pseudo-testcross strategy

Characterization of Polysaccharides Extracted from a Cultivated Brown Alga Costaria costata During the Harvest Period

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Journal of Ocean University of China 2018年第5期17卷 1209-1217页

作者： LIU Nannan WU Xin FU Xiaoting duan delin XU Jiachao GAO Xin College of Food Science and Engineering Ocean University of ChinaQingdao 266003China State Key Laboratory of Seaweed Bioactive Substances Qingdao 266000China Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China

In this study, variations in composition and properties of polysaccharides isolated from brown algae Costaria costata were analyzed. The algae were collected from May to July of the harvest period. The carbohydrates alginate and fucoidan were extracted with selected solvents. 1H Nuclear magnetic resonance and rheology were used to investigate the monomer composition and rheological characteristics of alginate. Gas chromatography and Fourier transform-infrared spectroscopy were employed to investigate the compositional properties of purified fucoidan. The results indicated that the composition and properties of alginate and fucoidan varied during the life of this alga. The alginate from the alga harvested in May and June had a higher molecular weight, viscosity, and proportion of mannuronic acid, whereas that harvested in July had a lower molecular weight and viscosity but a higher proportion of guluroinc acid. The alginate from C. costata had a higher molecular weight and a different mannuronic acid： guluronic acid ratios compared with other algae; thus, it could be used in the chemical, food, cosmetics, and pharmaceutical industries. Fucoidan content reached the maximum in June. Substantial changes in the molecular weight distribution, monosaccharide composition, and sulfate content occurred simultaneously. The fraction of fucose in the polysaccharides decreased significantly from June to July, whereas that of mannose increased. This alga can be harvested during different growth periods to obtain fucoidans and alginates with different compositions and, therefore, with different biological properties.

关键词： alginate brown alga Costaria costata fucoidan polysaccharides

Bioactive substances of various seaweeds and their applications and utilization

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Journal of Oceanology and Limnology 2019年第3期37卷 779-782页

作者： duan delin CRITCHLEY Alan T. FU Xiaoting PEREIRA Leonel Key Lab of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China Center for Ocean Mega-Science Chinese Academy of ScienceQingdao 266071China State Key Lab of Seaweed Bioactive Substances Qingdao 266400China Verschuren Centre for Sustainability in Energy and the Environment Cape Breton UniversitySydneyNova Scotia B1P 6L2Canada College of Food Science & Engineering Ocean University of ChinaQingdao 266000China MARE (Marine and Environmental Sciences Centre) Department of Life SciencesFaculty of Sciences and TechnologyUniversity of CoimbraCoimbra 3000-456Portugal

Over the past century or more,traditional uses of seaweeds began and initially comprised the simple collection,sorting and drying of seaweeds harvested from the wild,or cast upon the beach,for use as soil“manure/fertilizer/improver”.Some macroalgae were used for unprocessed food purposes and subsequently harvested and cultivated biomasses of selected brown and red seaweeds becameindustrially processed to yield alginates,mannitol,iodine,agar,agarose,carrageenan,and more recently biostimulants,*** from various seaweeds are very diverse and are commonly applied in a range that includes textiles and dying,food/feed and supplements and specialized medical applications.

关键词： century collection macroalgae

Low genetic diversity in the endangered marine alga Silvetia siliquosa(Ochrophyta:Fucaceae)and the implication to conservation

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Journal of Oceanology and Limnology 2022年第1期40卷 216-225页

作者： Yanshuo LIANG Jie ZHANG Xiaohan SONG Han-Gil CHOI Xu GAO delin duan Zi-min HU CAS and Shandong Province Key Laboratory of Experimental Marine Biology Center for Ocean Mega-ScienceInstitute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory for Marine Biology and Biotechnology Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China University of Chinese Academy Sciences Beijing 100049China Faculty of Biological Science and Institute for Environmental Science Wonkwang UniversityIksan 54538Korea Ocean School Yantai UniversityYantai 264005China

Although signifi cant research eff orts have been targeted toward conservation and management of endangered terrestrial fl ora and fauna,attempts have been limited to conserve threatened *** siliquosa is an ecologically and commercially vital brown alga that is uniquely distributed in the Yellow-Bohai Sea and along the southwest coast of Korea.A massive decline in its distribution range and biomass from the mid-1990s onward indicates that this species has become *** the present study,we used nuclear internal transcribed spacer and concatenated mitochondrial cytochrome oxidase I subunit+intergenic spacer to estimate the genetic diversity,population connectivity,and degree of genetic diff erentiation of *** in China and *** molecular results exhibited strikingly low levels of haplotype/ribotype and nucleotide diversity in *** populations,with only three mitochondrial haplotypes and nuclear ribotypes detected in 136 and 143 specimens,*** analysis of molecular variance revealed 85%-95%of genetic variance among *** diff erentiation coeffi cient(F_(ST))and gene fl ow(N_(m))suggested that two populations(JIN and GWA)along the southern coast of Korea are highly divergent from the others,with weak genetic *** signifi cant genetic diff erentiation was observed among populations either in China or along the geographically proximate west coast of ***,four independent management units were designated for sustainable management:the LII and RUS populations in China,the YEO and CHA populations along the west coast of Korea,and each of the GWA and JIN populations along the south coast of *** suggest that artifi cial cultivation and transplantation of *** are the eff ective approaches for restoration and conservation.

关键词： biodiversity conservation endangered seaweed genetic diversity habitat loss Silvetia siliquosa

Histo-blood group antigens in Crassostrea gigas and binding profiles with GⅡ.4 Norovirus

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Journal of Oceanology and Limnology 2018年第4期36卷 1383-1391页

作者： MA Liping LIU Hui SU Laij in ZHAO Feng ZHOU Deqing duan delin Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China Key Laboratory for Sustainable Utilization of Marine Fisheries Resources Ministry of AgricultureYellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdao 266071China Laboratory for Marine Biology and Biotechnology Qingdao National Laboratory for Marine Science and TechnologyQingdao 266071China

Noroviruses(NoVs) are the main cause of viral gastroenteritis outbreaks worldwide, and oysters are the most common carriers of NoV contamination and transmission. NoVs bind specifically to oyster tissues through histo-blood group antigens(HBGAs), and this facilitates virus accumulation and increases virus persistence in oysters. To investigate the interaction of HBGAs in Pacific oysters with GⅡ.4 NoV, we examined HBGAs with ELISAs and investigated binding patterns with oligosaccharide-binding assays using P particles as a model of five GⅡ.4 NoV capsids. The HBGAs in the gut and gills exhibited polymorphisms. In the gut, type A was detected(100%), whereas type Leb(91.67%) and type A(61.11%) were both observed in the gills. Moreover, we found that seasonal NoV gastroenteritis outbreaks were not significantly associated with the specific HBGAs detected in the oyster gut and gills. In the gut, we found that strain-2006 b and strain-96/96 US bound to type A and H1 but only weakly bound to type Leb; in contrast, the Camberwell and Hunter strains exhibited weak binding to types H1 and Ley, and strain-Sakai exhibited no binding to any HBGA type. In the gills, strain-96/96 US and strain-2006 b bound to type Leb but only weakly bound to type H1; strains Camberwell, Hunter, and Sakai did not bind to oyster HBGAs. Assays for oligosaccharide binding to GⅡ.4 NoV P particles showed that strain-95/96 US and strain-2006 b strongly bound to type A, B, H1, Leb, and Ley oligosaccharides, while strains Camberwell and Hunter showed weak binding ability to type H1 and Ley oligosaccharides and strain-Sakai showed weak binding ability to type Leb and Ley oligosaccharides. Our study presents new information and enhances understanding about the mechanism for NoV accumulation in oysters. Further studies of multiple NoV-tissue interactions might assist in identifying new or improved strategies for minimizing contamination, including HBGA-based attachment inhibition or depuration.

关键词： Crassostrea gigas norovirus histo-blood group antigen binding

Early development of Chondrus ocellatus Holm(Gigartinaceae,Rhodophyta)

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Chinese Journal of Oceanology and Limnology 2006年第2期24卷 129-133页

作者： WANG Aihua WANG Jicheng duan delin Institute of Oceanology The Chinese Academy of Sciences Qingdao 266071China The Graduate School of the Chinese Academy of Sciences Beijing 100039China

Chondrus is an economically important red algae widely used for food and biochemical *** early development is crucial for the culture and seedling *** chose tetraspores and carpospores of Chondrus ocellatus as examples for experiment of the culture,induction and release in laboratory condition,aiming to understand early development of *** and to apply in seedling *** *** were collected in Qingdao,China,from *** *** the gametophyte and tetrasporophyte were brushed and washed with sterilized seawater,the algal materials were treated in 1.5%KI for 20 min,then were dried for lh to stimulate the releasing of *** the spores released overnight,it were cultured in PES medium,incubated at 18℃,10±2μmol/(m^(2).s^(1))in 12:12h(light:dark).The observation and recording under microscope were carried *** observation of the early development showed that both tetraspore and carpospore are similar to each *** general,three stages of the early development were shown being division,discoid crust and seedling *** the division stage,the most obvious feature was the increasing of cell number;during the discoid crust stage,the discoid crust had a three-dimensional axis,and it began to differentiate into two types of cells:the basal cells and the apical cells;and to the seedling stage,several protuberances-like appeared on the discoid crusts and formed juvenile *** and tetraspores exhibited a similar development process that included division stage,discoid crust stage and seedling stage.

关键词： Chondrus ocellatus early development carpospore tetraspore discoid crust seedlings