海洋微生物来源的岩藻多糖降解酶

近年来研究发现,岩藻多糖及其降解产物具有多种重要的生物学活性,对岩藻多糖降解酶的关注日益增多。本文概述了海洋微生物来源的岩藻多糖降解酶的发现、活性检测方法、性质、应用等方面的研究进展。同时展望了现代组学及结构生物学技术快速发展对海洋微生物来源的新型岩藻多糖降解酶研究的推动作用。     

岩藻多糖在2型糖尿病大鼠模型的作用及机制研究

目的:研究岩藻多糖(Fucoidan)对链脲佐菌素诱导实验性2型糖尿病大鼠的治疗作用.方法:在SD大鼠建立2型糖尿病动物模型,随机分为5组,分别用生理盐水、岩藻多糖高、中、低剂量(600mg/kg、400mg/kg、150mg/kg)、二甲双胍(200mg/kg)灌胃给药30天,同时采用正常SD大鼠为正常对照.测定各组大鼠体重、空腹血糖、血脂和血清胰岛素水平.结果:高、中剂量岩藻多糖能够明显降低模型动物空腹血糖水平,降低空腹胰岛素水平,改善胰岛素抵抗,降低血清胆固醇、甘油三酯、低密度脂蛋白胆固醇含量(P<0.05),升高高密度脂蛋白胆固醇含量(P<0.05),其作用效果与阳性对照药二甲双胍效果相当.低剂量岩藻多糖作用效果不明显.结论:岩藻多糖有降血糖、调节血脂紊乱的功能,时实验性2型糖尿病大鼠有治疗作用.     

岩藻多糖改善胃部疾病的研究进展

岩藻多糖具有多种生物活性,能改善胃部炎症,缓解胃癌化疗副作用,在改善胃部疾病方面具有一定的健康功效,在食品、药品的研发中具有广阔的发展前景。本文综述了岩藻多糖在改善胃部疾病方面的研究进展。     

过硫酸化岩藻多糖硫酸酯

岩藻多糖硫酸酯(fucoidan或fucan sulfate,FS)是一种硫酸多糖,具有多种生物活性。硫酸基团对多糖活性起重要作用,并且多糖活性与硫酸基团含量呈正相关。近些年,国内外研究者对SF进行硫酸化修饰以获得高硫酸基团含量和高活性的过硫酸化岩藻多糖硫酸酯。为促进对FS硫酸化修饰的深入了解,本文对FS活性与硫酸基团含量及位置的关系、过硫酸化方法及过硫酸化岩藻多糖硫酸酯活性的研究进展进行了综述。     

黄杆菌菌株RC2-3基因组生物信息学分析及高效降解岩藻多糖功能基因的挖掘

以从海带中筛选获得的一株具有降解岩藻多糖能力的黄杆菌菌株RC2-3为研究对象，该菌株产的岩藻多糖酶可以高效降解不同来源的岩藻多糖。为进一步探究菌株RC2-3降解岩藻多糖的机制，推动岩藻寡糖的酶法生产，采用Illumina测序技术对菌株RC2-3进行基因组测序、基因功能注释和碳水化合物活性酶注释以及岩藻多糖降解相关基因的生物信息学分析。结果表明，黄杆菌菌株RC2-3基因组全长3 414 532 bp,共编码2 967个基因，GC含量为30.92%。经碳水化合物活性酶数据库注释获得213个基因，与岩藻多糖降解有关的包括7个岩藻糖结合结构域的基因；2个β-D-岩藻糖苷酶(EC 3.2.1.38)基因；2个属于GH141家族的α-L-岩藻糖苷酶(EC 3.2.1.51)基因；12个属于GH29家族的α-1, 3/1, 4-L-岩藻糖苷酶(EC 3.2.1.111)基因；9个属于GH95家族的α-1, 2-L-岩藻糖苷酶(EC 3.2.1.63)基因。此外，通过与已报道的蛋白序列比对发现，岩藻多糖酶基因RC2.3＿GM001247编码的蛋白序列与FunA蛋白序列同源性达到70.98%,岩藻多糖酶...     

扁藻多糖的性质及结构特征的研究

利用不同浓度范围的乙醇从培养至平衡期后的扁藻细胞内沉淀分离出不同组分的扁藻多糖级分,对其理化性质和结构特征进行了相应的测定和对比分析。结果表明,经过凝胶层析分离后,乙醇浓度在0%~50%范围时沉淀的扁藻多糖与乙醇浓度为50%~75%范围时沉淀的扁藻多糖相比,前者分子量大、糖含量高、其结合的蛋白质也多,但前者的溶解度和粘度小于后者。两个级分的扁藻多糖的官能团相似,均含有硫酸基和氨基,但其含量不同。所得结果为深入研究扁藻多糖的结构及其实际应用提供了实验依据。     

HILIC-ESI-HCD-MS/MS寡糖图谱分析用于海参褐藻糖胶的结构鉴定

褐藻糖胶也称为岩藻黄质、岩藻多糖、褐藻多糖,是一种硫化的多糖,常见于海参、海藻等褐藻纲及海藻类的植物中。研究发现,具有特定硫酸化结构的糖类结构域在生物功能中起着至关重要的作用。因此,对褐藻糖胶寡糖进行精细的结构表征,尤其是确定其硫酸化结构,对于理解褐藻糖胶的结构-功能关系至关重要。     

金属硫蛋白及其在心血管系统疾病发生中的作用

金属硫蛋白（MT）是一类富含半胱氨酸的低分子量非酶蛋白,具有重金属解毒、调节微量元素代谢、清除自由基抗氧化等多种生物学功能。MT可作为高效的内源性细胞保护剂,对多种心血管系统疾病具有重要的保护作用。简要综述了MT的理化性质及生物学特性,MT在心肌缺血再灌损伤、心力衰竭、阿霉素诱发的心脏毒性、高血压及动脉粥样硬化等心血管疾病发病过程中的作用及其可能的机制。     

热休克蛋白的分子生物学

热休克应答是一种普遍的生物学现象。热休克蛋白(hsp)分为高分子量和低分子量两大类。hsp基因的表达是在转录水平和翻译水平进行调节的。hsp的功能与调节机体的耐热性有关。     

抗菌肽及其重组表达系统研究进展

抗菌肽作为一类具有多种生物学功能的小肽,具有分子量小、不含外源成分、不易产生耐药性等特性,成为近年来的研究热点。该文综述了抗菌肽的种类、作用机制及其重组表达系统等方面的研究进展,并对存在的热点问题进行了探讨。     

Melanogenesis inhibitory effect of low molecular weight fucoidan from <Emphasis Type="Italic">Undaria pinnatifida</Emphasis>

In this study, fucoidans with different molecular weight that were isolated from the brown alga Undaria pinnatifida (Phaeophyceae, Laminariales) were investigated for their ability to inhibit melanogenesis and scavenge superoxide and hydroxyl radicals. Fucoidan samples with low molecular weights of 89, 35, 17, and 6 kDa were prepared by radiation-degradation of a 378 kDa fucoidan isolated from U. pinnatifida. The inhibitory activity of fucoidan against melanin biosynthesis in B16BL6 melanoma cells was enhanced for low molecular weight samples. To investigate the increase in melanogenesis inhibition exhibited by the low molecular weight fucoidan, tyrosinase inhibition activity and radical scavenging activities were measured. There was an increase in the tyrosinase inhibition activity for low molecular weight samples. Additionally, the radical scavenging activity was increased for lower molecular weight fucoidans. These results suggest that low molecular weight fucoidans from seaweeds may have beneficial biological properties.     

Microbial fucoidan degradation by Luteolibacter algae H18 with deacetylation

A bacterial strain that assimilates fucoidan from Cladosiphon okamuranus as sole carbon source was isolated as Luteolibacter algae H-18. It was found that it degraded fucoidan by intracellular enzymes, and that the degradation reactions were catalyzed by multiple enzymes. One enzyme, designated fraction B, was established to exhibit the deacetylation reaction of fucoidan. Other enzyme(s), designated fraction A, catalyzed the reaction(s) lowering the molecular weight of fucoidan.     

Microbial Fucoidan Degradation by Luteolibacter algae H18 with Deacetylation

A bacterial strain that assimilates fucoidan from Cladosiphon okamuranus as sole carbon source was isolated as Luteolibacter algae H-18. It was found that it degraded fucoidan by intracellular enzymes, and that the degradation reactions were catalyzed by multiple enzymes. One enzyme, designated fraction B, was established to exhibit the deacetylation reaction of fucoidan. Other enzyme(s), designated fraction A, catalyzed the reaction(s) lowering the molecular weight of fucoidan.     

Fucoidan exerts antidepressant-like effects in mice via regulating the stability of surface AMPARs

The inflammatory hypothesis is one of the most important mechanisms of depression. Fucoidan is a bioactive sulfated polysaccharide abundant in brown seaweeds with anti-inflammatory activity. However, the antidepressant effects of fucoidan on chronic stress-induced depressive-like behaviors have not been well elucidated. Here, we used two different depressive-like mouse models, lipopolysaccharide (LPS) and chronic restraint stress (CRS) models, to explore the detailed molecular mechanism underlying its antidepressant-like effects in C57BL/6J mice by combining multiple behavioral, molecular and immunofluorescence experiments. Adenovirus-mediated overexpression of caspase-1 and pharmacological inhibitors were also used to clarify the antidepressant mechanisms of fucoidan. We found that acute administration of fucoidan did not produce antidepressant effects in the tail suspension test (TST) and forced swim test (FST). Interestingly, chronic fucoidan administration not only dose-dependently reduced stress-induced depressive-like behaviors in the TST, FST, sucrose preference test (SPT), and novelty-suppressed feeding test (NSFT), but also alleviated the downregulation of brain-derived neurotrophic factor (BDNF)-dependent synaptic plasticity via inhibiting caspase-1-mediated inflammation in the hippocampus of mice. Moreover, fucoidan significantly ameliorated behavioral and synaptic plasticity abnormalities in the overexpression of caspase-1 in the hippocampus of mice. Furthermore, blocking BDNF abolished the antidepressant-like effects of fucoidan in mice. Therefore, our findings clearly indicate that fucoidan provides a potential supplementary noninvasive treatment for depression by inhibition of hippocampal inflammation.     

Characterization of sugar binding by osteoclast inhibitory lectin

Osteoclast inhibitory lectin (OCIL) is a membrane-bound C-type lectin that blocks osteoclast differentiation and, via binding to its cognate receptor NKRP1D, inhibits natural killer cell-mediated cytotoxicity. OCIL is a member of the natural killer cell receptor C-type lectin group that includes CD69 and NKRP1D. We investigated carbohydrate binding of soluble recombinant human and mouse OCIL in enzyme-linked immunosorbent assay-based assays. OCIL bound immobilized high molecular weight sulfated glycosaminoglycans, including fucoidan, lambda-carrageenan, and dextran sulfate, but not unsulfated dextran or sialated hyaluronic acid. Carbohydrate binding was Ca(2+)-independent. Binding of immobilized low molecular weight glycosaminoglycans, including chondroitin sulfate (A, B, and C forms) and heparin, was not observed. However, the soluble forms of these low molecular weight glycosaminoglycans competed for OCIL binding of immobilized fucoidan (as did soluble fucoidan, dextran sulfate, and lambda-carrageenan), indicating that OCIL does recognize these carbohydrates. Inhibition constants for chondroitin sulfate A and heparin binding were 380 and 5 nm, respectively. Immobilized and soluble monosaccharides did not bind OCIL. The presence of saturating levels of fucoidan, dextran sulfate, and lambda-carrageenan did not affect OCIL inhibition of osteoclast formation. The fucoidan-binding lectins Ulex europaeus agglutinin I and Anguilla anguilla agglutinin did not block osteoclast formation or affect the inhibitory action of OCIL. Although the osteoclast inhibitory action of OCIL is independent of sugar recognition, we have found that OCIL, a lectin widely distributed, but notably localized in bone, skin, and other connective tissues, binds a range of physiologically important glycosaminoglycans, and this property may modulate OCIL actions upon other cells.     

Purification and characteristics of fucoidanase obtained from <Emphasis Type="Italic">Dendryphiella arenaria</Emphasis> TM94

Dendryphiella arenaria TM94 is an obligate marine fungus. Fucoidanase expressed by TM94 by solid state fermentation was purified. The fermented solid medium was extracted with citric acid buffer, and the extracts were precipitated by acetone and separated on Sephadex G-100 chromatography. The specific fucoidanase activity of purified enzyme was 27-fold than that of the crude enzyme. The recovery of the enzyme was 17.69%. SDS-PAGE was used to identify the purity and the molecular weight of the fucoidanase. A single band appeared on SDS-PAGE gel which suggested that relatively pure fucoidanase has been obtained. The molecular weight of fucoidanase is 180 kDa and the isoelectric point was about pH 4.4. The purified fucoidanase appeared to have the maximum enzymatic activity at pH 6.0. K_M and the maximum velocity of the enzyme was 6.56 mg·mL⁻¹ and 6.55 mg·mL⁻¹·min⁻¹ by using fucoidan from Fucus vesiculosus as substrate. The enzyme may be a type of endo-fucoidanase which could hydrolyze high molecular weight fucoidan to low molecular weight fucoidan rather than to fucose.     

Interaction of fucoidan with the proteins of the complement classical pathway

Fucoidan inhibits complement by mechanisms that so far remain to be unraveled, and the objective of this work was to delineate the mode of inhibition by this sulfated polysaccharide. For that purpose, low molecular weight fractions of algal (Ascophyllum nodosum) fucoidan containing the disaccharide unit [-->3)-alpha-L-Fuc(2SO3(-))-(1-->4)-alpha-L-Fuc(2,3diSO3(-))-(1-->](n) have been studied. Gel co-affinity electrophoresis and a new affinity capillary electrophoresis (ACE) method have been implemented to characterize fucoidan-complement protein complexes. Fucoidan binds C1q, likely to its collagen-like region through interactions involving lysine residues, and then prevents the association of the C1r(2)-C1s(2) subunit, required to form the fully active C1. In addition to C1q, fucoidan forms a complex with the protein C4 as observed by ACE. The fucoidan inhibits the first steps of the classical pathway activation that is of relevance in view of the proinflammatory effects of the subsequent products of the cascade. This study shows that a high level of inhibitory activity can be achieved with low molecular weight carbohydrate molecules and that the potential applicability of fucoidan oligosaccharides for therapeutic complement inhibition is worthy of consideration.     

Fucoidan isolated from the popular edible brown seaweed Sargassum fusiforme suppresses lipopolysaccharide-induced inflammation by blocking NF-κB signal pathway

Journal of Applied Phycology - The anti-inflammatory effect of a fucoidan with a molecular weight of 102.67&nbsp;kDa isolated from an enzymatic digest of Sargassum fusiforme was investigated in...     

Polysaccharides composition from tropical brown seaweeds

Polysaccharides composition of the tropical brown seaweeds Turbinaria turbinata, Sargassum filipendula, Dictyota caribaea and Padina perindusiata collected at Yucatan Peninsula (Mexico) was determined in this study. Crude fucoidan extracted with HCl and alginate extracted with a hot alkali solution were characterized in terms of their molecular weight, sulfate content, uronic acid, total carbohydrate and neutral sugar components. Low molecular weight sulfated‐fucoidan was the major component in all species studied. Fucoidan from T. turbinata and from D. caribaea were characterized as a homofucan, with fucose as the neutral sugar. Fucoidan from S. filipendula was composed of a galactofucan, and fucoidan from P. perindusiata was characterized as a heterofucan consisting of fucose, glucose and galactose. The Fourier transform infrared (FT‐IR) spectra of fucoidan extracted from species studied indicated that the majority of sulfate groups are located at C‐4 and to a lesser extent at C‐2 and/or C‐3 of the fucopyranose residues. This could be advantageous since several therapeutic effects have been reported for fucoidans with similar characteristics. FT‐IR spectra from D. caribaea and P. perindusiata revealed the presence of O‐acetyl groups in crude fucoidan, which could be potentially utilized as an immune stimulant. Molecular weight of alginate varied between 595 and 1301 kDa with similar uronic acid content in all species. Alginate M : G ratio inferred from FT‐IR spectra suggests a high content of G‐block in all species. Potential applications of these polysaccharides are discussed.     

Mild acid hydrolysis of fucoidan: characterization by electrophoresis and FT-Raman spectroscopy

Along with proteins, lipids, water and minerals, polysaccharides are the main chemical compounds of which macroalgae are built. Among the chemical compounds now widely examined is fucoidan (fucan, fucosan, sulfate fucan or sulfated fucan), a fucose-containing sulfated polysaccharide. Fucoidans isolated from different species have been extensively studied because of their varied biological properties, including anticoagulant and antitumor effects. Methodology based on mild acid hydrolysis can be used as an efficient tool to study the relationship between molecular weight of the sulfated polysaccharides and their biological activities. Anticancer activity of fucoidans can be significantly enhanced by lowering their molecular weight only when they are depolymerized under mild conditions. In this study, fucoidan was identified during extraction with H₂SO₄ and HCl; its presence was confirmed by FT-Raman spectroscopy in aqueous solution. In particular, shifts at 840 cm⁻¹ were analysed, which are due to the presence of sulfate at the axial C-4 position, as were the shifts at about 811–809 cm⁻¹, for which the sulfated fucoidan is responsible. Shifts of electrophoretic bands of fucoidan resulting from mild acid hydrolysis in H₂SO₄ and HCl were also analysed. The analytical procedure was developed using apparatus for cellulose acetate membrane electrophoresis and this was supplemented by semi-quantitative analysis.