用冻融结合超声波法辅助提取浒苔多糖的工艺条件

浒苔(Enteromorpha prolifera)多糖具有多种生物学活性,一种高效并对生物学活性影响小的多糖提取工艺对其生物学活性研究有重要意义。用冻融结合超声波法研究浒苔多糖的提取工艺,在单因素实验的基础上,选择水料质量比、超声波功率、提取时间、冻融-超声波次数进行4因素3水平的正交实验提取浒苔多糖。结果表明:浒苔多糖最佳提取条件为水料质量比55、超声波功率600 W、超声波作用时间8 min、冻融-超声波2次,浒苔多糖得率为19.124%。     

Biodegradation of <Emphasis Type="Italic">Enteromorpha</Emphasis> polysaccharides by intestinal micro-community from <Emphasis Type="Italic">Siganus oramin</Emphasis>

Micro-communities are supposed to have more potential functions of biodegradation of polysaccharides than single strain; however, the intestinal micro-communities involved in the biodegradation of Enteromorpha polysaccharides (EP) were seldom reported. In order to obtain the EP-degrading micro-community, the intestines of Siganus oramin was obtained to isolate the micro-communities, which were enriched by 0.3% of EP as the sole carbon source. A stable micro-community with EP degradative capability was achieved after seven generations of subculture, named H1. Results showed that H1 was able to degrade 75% of EP within 24 hours, and the activity of EP lyases reached 500 U mL^?1 in 32 hours. With denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analysis, ten bacteria closely related to Marinomonas pontica, Microbacterium sp., Leucobacter chironomi, Cyclobacterium sp., Algoriphagus winogradskyi, Pseudoalteromonas sp. and Vibrio sp. were determined. Furthermore, compared with the DGGE bands sequence and the clone library analysis, the dominant bacteria of the EP-biodegrading micro- community were Pseudoalteromonas sp. and Vibrio sp., with the respective proportion of 38% and 46%, and they should play an important role in EP degradation together with other degrading bacteria in the micro-community H1.     

Purification and characterization of iron-cofactored superoxide dismutase from Enteromorpha linza

A superoxide dismutase was purified from Enteromorpha linza using a simple and safe procedure, which comprised phosphate buffer extraction, ammonium sulphate precipitation, ion exchange chromatography on Q-sepharose column, and gel filtration chromatography on Superdex 200 10/300GL. The E. linza superoxide dismutase （E/SOD） was purified 103.6-fold, and a yield of 19.1% and a specific activity of 1 750 U/rag protein were obtained. The SDS-PAGE exhibited E/SOD a single band near 23 kDa and the gel filtration study showed E/SOD＇s molecular weight is near 46 kDa in nondenatured condition, indicating it＇s a homodimeric protein. E/SOD is an iron-cofactored superoxide dismutase （Fe-SOD） because it was inhibited by hydrogen peroxide, insensitive to potassium cyanide. The optimal temperature for its maximal enzyme activity was 35℃, and it still had 29.8% relative activity at 0℃, then E/SOD can be classified as a cold-adapted enzyme. E/SOD was stable when temperature was below 40℃ or the pH was within the range of 5 10. The first 11 N-terminal amino acids orE/SOD were ALELKAPPYEL, comparison of its N-terminal sequence with other Fe-SOD N-terminal sequences at the same position suggests it is possibly a chloroplastic Fe-SOD.     

湛江海域浒苔属Enteromorpha种类的形态与显微结构

用比较解剖学方法观察提江沿海浒苔属（Enteromorpha ）海藻的形态结构,并结合统计分析对其进行 物种鉴定。结果表明,依据藻体分校的有元、数量及分校与主枝直径的大小等形态特征,鉴定的浒苔属j海藻有6种1个变型,分别为缘管浒苔（E. linza）、肠浒苔（ E. intestinalis ）、扁浒苔（ E. compressa ） ,浒苔（E. proliferα）、 条浒苔（E. clathratα）、由浒苔（ E·flexω5α）和肠浒苔宽叶变型（ E. intestinalis f.broadifolium） 0藻体长度由 大到小依次为浒苔、条浒苔、缘管浒苔、肠浒苔、扁浒苔和炀浒苔宽叶变型：细胞呈多角形,藻体的位置不同其细胞大小有所变化,除由浒苔和条浒苔外,其他4种1个变型浒苔的中部细胞最小;所有种类的细胞均有淀粉核,其中肠浒苔、缘管浒苔和扇浒苔仅1个淀粉核,而其他种出现多个淀粉核,尤其条浒苔和曲浒苔数量最多,可达6个以上。定量描述浒苔属6种1个变型的形态特征、内部结构,并发现浒苔属海藻非中空营状的结构特征。     

Effect of polysaccharide from Enteromorpha prolifera on maize seedlings under NaCl stress

In this study,a polysaccharide from Enteromorpha prolifera(EP)was extracted and its effect on maize seedlings under NaCl stress was investigated.Firstly,the components and structure of the EP were determined.We found that EP is a sulfated polysaccharide of high-molecular weight(Mw,1 840 KDa)heteropoly saccharides and the main mono saccharide is rhamno se.The poly saccharide was applied to explore its effect on the growth of maize seedlings and its defense response under a salt stress.The results show that EP could promote the growth of maize seedlings under the salt stress.In addition,EP was shown able to significantly regulate membrane permeability and adjustment of osmotic substances such as soluble protein,soluble sugar,and proline,antioxidant enzymes containing superoxide dismutase,catalase,peroxidase,and ascorbate peroxidase.Therefore,EP is an effective salt-resistant substance for the growth of maize seedlings under NaCl stress.     

在栽培细基江篱繁枝变种的水塘中用百草枯清除浒苔试验

利用半咸淡水塘栽培细基江繁枝变种,已成为海水养殖事业中的重要组成部分。但目前遇到最难处理的问題是故害藻类的緣藻类浒苔大量繁殖,严重地影响了江篱的生产。为了清除浒苔,文章作者在海南省海口市附近的水塘中进行了用百草枯清除浒苔的试验。结果证明在放干水的塘底直接用3‰-6‰的百草枯溶液噴撤在江篱和浒苔藻体上,一小时后,再放进海水,则浒苔全部死亡,而江篱仍然很茂盛地生长。     

Selenylation Modification of Degraded Polysaccharide from Enteromorpha prolifera and Its Biological Activities

Polysaccharide extracted from Enteromorpha prolifera possessed excellent biological activities,but its molecular weight was greatly high which influenced the activity.Organic Se had higher biological activities and was safer than inorganic Se species.In the present study,Enteromorpha polysaccharide was degraded to low molecular weight by free-radical degradation method of H_2O_2 and ascorbic acid.By single factor and orthogonal experiments,the optimal degradation conditions were reaction time of 2 h,reaction temperature of 50℃,H_2O_2/ascorbic acid(n/n=1:1)concentration of 15 mmol L~(-1),and solid-liquid ratio of 1:50(g mL~(-1)).Then,the degraded polysaccharide was chemically modified to obtain its selenide derivatives by nitric acid-sodium selenite method.The selenium content was 1137.29μg g~(-1),while the content of sulfate radical had no change.IR spectra indicated that the selenite ester group was formed.Degraded polysaccharide selenide was characterized and evaluated for antioxidant,antifungal and antibacterial activities.The results showed that degraded polysaccharide selenide had strong capacity of scavenging DPPH and·OH free radical.It had significant antibacterial properties for Escherichia coli,Bacillus subtilis and Salmonella spp.,and it also had significant antifungal properties for Apple anthrax.The result ascertained degradation and selenylation modification did not change the main structure of polysaccharides.It was possible that free-radical degradation was an effective way for enhancing antioxidant activity to decrease molecular weight of polysaccharides.     

响应面法优化波吉卵囊藻多糖提取条件

【目的】探究波吉卵囊藻(Oocystis borgei)多糖提取的最优条件。【方法】通过单因素和响应面实验,考察不同提取温度、氢氧化钠浓度、提取时间对波吉卵囊藻多糖提取率的影响。【结果】各因素对波吉卵囊藻多糖提取率的影响显著性表现为氢氧化钠浓度提取温度提取时间;最佳提取条件:提取温度58.5℃、氢氧化钠浓度0.305mol/L、提取时间68 min,此时提多糖取率为11.86%,与模型预测值11.93%接近。【结论】采用响应面法建立的模型可以较好地预测提取温度、氢氧化钠浓度和提取时间对波吉卵囊藻多糖的提取率的影响。     

Molecular diversity of Enteromorpha from the coast of Yantai: a dual-marker assessment

We collected nine Enteromorpha specimens from the coast of Yantai and evaluated their diversity based on analyses of their ITS （internal transcribed spacer） and 5S rDNA NTS （non-transcribed spacer） sequences. The ITS sequences showed slight nucleotide divergences between Enteromorpha linza and Enteromorpha prolifera. In contrast, multiple highly variable regions were found in the ITS region of Enteromorpha flexuosa. In general, there were more variable sites in the NTS region than in the ITS region in the three species. The variations in 5S rDNA NTS sequences indicated that the molecular diversity of Enteromorpha from the coast of Yantai is very high. However, a phylogenetic tree constructed using 5S rDNA NTS sequence data indicated that genetic differences were not directly related to geographical distribution.     

Erratum to: Purification and characterization of iron-cofactored superoxide dismutase from Enteromorpha linza

<正>Erratum to:Chinese Journal of Oceanology and Limnology Vol.31 No.6,P.1190-1195,2013http://dx.doi.org/10.1007/s00343-013-3049-3The supporting grant information on the footnote of the original version of this article needs to be altered and updated.The new information is given below.     

Optimization of dilute acid hydrolysis of Enteromorpha

Acid hydrolysis is a simple and direct way to hydrolyze polysaccharides in biomass into fermentable sugars. To produce fermentable sugars effectively and economically for fuel ethanol, we have investigated the hydrolysis of Enteromorpha using acids that are typically used to hydrolyze biomass:H2SO4, HCl, H3PO4 and C4H4O4 (maleic acid). 5%(w/w) Enteromorpha biomass was treated for different times (30, 60, and 90 min) and with different acid concentrations (0.6, 1.0, 1.4, 1.8, and 2.2%, w/w) at 121°C. H2SO4 was the most effective acid in this experiment. We then analyzed the hydrolysis process in H2SO4 in detail using high performance liquid chromatography. At a sulfuric acid concentration of 1.8% and treatment time of 60 min, the yield of ethanol fermentable sugars (glucose and xylose) was high, (230.5 mg/g dry biomass, comprising 175.2 mg/g glucose and 55.3 mg/g xylose), with 48.6% of total reducing sugars being ethanol fermentable. Therefore, Enteromorpha could be a good candidate for production of fuel ethanol. In future work, the effects of temperature and biomass concentration on hydrolysis, and also the fermentation of the hydrolysates to ethanol fuel should be focused on.     

Chemical characteristics and anticoagulant activities of two sulfated polysaccharides from Enteromorpha linza (Chlorophyta)

Two sulfated polysaccharides, designated MP and SP, were extracted from the marine green alga Enteromorpha linza using hot water and then purified using ion-exchange and size-exclusion chromatography. The anticoagulant activities of MP and SP were examined by determination of their activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) using human plasma. Results showed that MP and SP were composed of abundant rhamnose with small amounts of xylose and glucuronic acid, whereas SP also contained a small amount of galactose. Approximate molecular weights of MP and SP were 535 and 502 kDa, respectively. As compared with SP, MP had higher contents of sulfate ester (19.0%) and uronic acid (14.9%). The MP mainly consisted of (1→4)-linked rhamnose residues with partially sulfated groups at the C-3 position, and small amounts of (1→3, 4)-linked rhamnose, (1→2, 4)-linked rhamnose, (1→4)-linked glucuronic acid and (1→4)-linked xylose residues. The SP contained abundant (1→4)-linked rhamnose with minor amounts of (1→3)-linked rhamnose, (1→3, 4)-linked rhamnose, (1→2, 4)-linked rhamnose, (1→4)-linked glucuronic acid, (1→4)-linked xylose, and (1→3)-linked galactose residues. The sulfate groups were mainly located at C-3 of (1→4)-linked rhamnose residues. Both MP and SP, in particular the former, effectively prolonged APTT and TT. This work demonstrates that MP and SP have unique structural characteristics distinct from those of other sulfated polysaccharides from Enteromorpha. The MP is a potential source of anticoagulant, and the difference in anticoagulant activities of the two sulfated polysaccharides is directly linked to the discrepancy of their chemical features.     

Erratum to: Molecular diversity of Enteromorpha from the coast of Yantai: a dual-marker assessment

<正>Erratum to:Chinese Journal of Oceanology and Limnology Vol.31 No.6,P.1210-1215,2013http://dx.doi.org/10.1007/s00343-013-3045-7The supporting grant information on the footnote of the original version of this article needs to be altered and updated.The new information is given below.     

Polyphenols Extracted from Enteromorpha clathrata Induce Apoptosis in Hepa1-6 Cell by Activating the Mitochondrial Apoptosis Signaling Pathways

Green alga Enteromorpha clathrata(E. clathrata) contains a variety of bioactive compounds, including polysaccharides,polyphenols and fat-soluble pigments etc., among which polyphenols exhibit a wide range of medicinal properties. E. clathrata polyphenols(ECPs) have shown various biological activities such as antioxidant, anti-inflammatory and antidiabetic effects; however, the potential of ECPs as an anti-cancer reagent remains unclear. The aim of this study was to investigate the anti-tumor act...     

Molecular phylogenetic analysis of attached Ulvaceae species and free-floating Enteromorpha from Qingdao coasts in 2007

Based on the sequence data of the nuclear ribosomal DNA internal transcribed spacer(ITS) 1,5.8 S,and ITS 2,the molecular phylogeny was analyzed on Ulvaceae species collected from Qingdao coasts in summer of 2007,including 15 attached Ulva and Enteromorpha samples from 10 locations and 10 free-floating Enteromorpha samples from seven locations.The result supported the monophyly of all free-floating Enteromorpha samples,implying the unialgal composition of the free-floating Enteromorpha,and the attached Ulvaceae species from Qingdao coasts were grouped into other five clades,suggesting that they were not the biogeographic origin of the free-floating Enteromorpha in that season.     

Preference of the herbivorous marine teleost Siganus canaliculatus for different macroalgae

The decomposition of a large amount of unexploited macroalgal resource along the coast of China often results in heavy environmental pollution. In order to pave a way of using macroalgae as the dietary ingredient of rabbitfish Siganus canaliculatus, one of a few farmed herbivorous marine teleosts in China, its preference （feeding selectivity） for different macroalgae was determined in this study. Seven seaweed species abundantly inhabiting the coast of east Guangdong Province were exposed simultaneously to rab-bitfish juveniles in laboratory （multiple-choice feeding） with their content and absolute intake assayed. It was found that the most preferred algae were Ulva prolifera, Gracilaria lemaneiformis and Chaetomorpha linum, less preferred algae were U. pertusa and Porphyra haitanensis, and least preferred ones were Sargassum fusiforme and Corallina sessilis. Such an order did not change when one to four relatively preferred seaweeds were removed. The preferred seaweeds were richer in protein and soluble sugar thus higher in energy than the least preferred. In addition, this fish was found to favor filamentous and flat algae rather than calcified ones. Ac-cordingly, the richness of nutrients and morphological characteristics determined the preference of S. canaliculatus for tested macro-algae.     

A Comparative Analysis of Four Kinds of Polysaccharides Purified from Furcellaria lumbricalis

1 Introduction Carrageenan, a water-soluble sulfated polymer, usually extracted from red algae, has been widely used in food industry due to its unique properties, such as gelation, emulsification, protein stabilizer, and so on. There are at least 13 types of carrageenans defined structurally based on the content of sulfate, galactose (Gal) and 3,6-anhydro-galactose (AnG), but only three types of them are commercially available, i.e., κ-, ι- and λ-type carrageenan. Different red algae prod…     

Overview of In-Situ Biodegradation and Enhancement

ＩｎｔｒｏｄｕｃｔｉｏｎＲｅｍｅｄｉａｔｉｏｎｂｙｎａｔｕｒａｌａｔｔｅｎｕａｔｉｏｎ (ＲＮＡ)ｉｓｔｈｅｐｒｅｆｅｒｒｅｄｍｅｔｈｏｄｆｏｒｒｅｍｅｄｉａｔｉｎｇｇｒｏｕｎｄｗａｔｅｒｃｏｎｔａｍｉ ｎａｔｅｄｂｙｔｈｅａｒｏｍａｔｉｃｆｕｅｌｈｙｄｒｏｃａｒｂｏｎｓ ,ｂｅｎｚｅｎｅ ,ｔｏｌｕｅｎｅ ,ｅｔｈｙｌｂｅｎｚｅｎｅ ,ａｎｄｘｙｌｅｎｅ (ＢＴＥＸ)ｅｔｃ .(Ｊｅｆ ｆｒｅｙ ,2 0 0 1) .ＢｕｔｔｈｅｒｅａｒｅｃｅｒｔａｉｎｃｏｎｄｉｔｉｏｎｓｕｎｄｅｒｗｈｉｃｈＲＮＡｉｓｉｎａｄｅｑｕａｔｅ…     

Ultrasonic Extraction of Polysaccharides from Laminaria japonica and Their Antioxidative and Glycosidase Inhibitory Activities

In the present study, ultrasonic extraction technique(UET) is used to improve the yield of polysaccharides from Laminaria japonica(LJPs). And their antioxidative as well as glycosidase inhibitory activities are investigated. Box-Behnken design(BBD) combined with response surface methodology(RSM) is applied to optimize ultrasonic extraction for polysaccharides. The optimized conditions are obtained as extraction time at 54 min, ultrasonic power at 1050 W, extraction temperature at 80℃ and ratio of material to solvent at 1:50(g m L-1). Under these optimal ultrasonic extraction conditions, an actual experimental yield(5.75% ± 0.3%) is close to the predicted result(5.67%) with no significant difference(P 0.05). Vitro antioxidative and glycosidase inhibitory activities tests indicate that the crude polysaccharides(LJP) and two major ethanol precipitated fractions(LJP1 and LJP2) are in a concentration-dependent manner. LJP2(30%-60% ethanol precipitated polysaccharides) possesses the strongest α-glucosidase inhibitory activity and moderate scavenging activity against hydroxyl radicals(66.09% ± 2.19%, 3.0 mg m L-1). Also, the inhibitory activity against α-glucosidase(59.08% ± 3.79%, 5.0 mg m L-1) is close to that of acarbose(63.99% ± 3.27%, 5.0 mg m L-1). LJP1(30% ethanol precipitated polysaccharides) exhibits the strongest scavenging activity against hydroxyl radicals(99.80% ± 0.00%, 3.0 mgm L-1) and moderate α-glucosidase inhibitory activity(47.76% ± 1.92%, 5.0 mg m L-1). LJP shows the most remarkable DPPH scavenging activity(66.20% ± 0.11%, 5.0 mg m L-1) but weakest α-glucosidase inhibitory activity(37.77% ± 1.30%, 5.0 mg m L-1). However, all these LJPs exert weak inhibitory effects against α-amylase. These results show that UET is an effective method for extracting bioactive polysaccharides from seaweed materials. LJP1 and LJP2 can be developed as a potential ingredient in hypoglycemic agents or functional food for the management of diabetes. This study provides scientific evidence and advances in the preparation technology and a hypoglycemic activities evaluation method for seaweed polysaccharides, especially glycosidase inhibition in combination with an antioxidative activity evaluation method.