黄芪多糖的超声波-闪式协同提取工艺优化及抗氧化活性研究

以黄芪为原料,采用超声波-闪式协同提取黄芪多糖。以黄芪多糖得率为评价指标,采用单因素试验和正交试验,确定最佳提取工艺。结果表明,超声波提取最佳工艺条件为：料液比1∶30（g/mL）,超声温度70 ℃,超声时间40 min,超声波功率500 W,在此条件下,黄芪多糖得率为7.11%;在对超声波提取工艺优化的基础上,协同闪式提取,最佳闪式提取工艺参数为：提取次数3 次,闪提时间100 s,提取电压160 V,在此条件下,黄芪多糖得率为11.03%;体外抗氧化活性试验结果表明：当黄芪多糖质量浓度为1.0 mg/mL 时,其对1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和2,2-联氨-双-3-乙基苯并噻唑啉-6-磺酸（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS）阳离子自由基的清除率分别为66.27%和58.27%。本研究获得了黄芪多糖的最佳提取方法,得到的黄芪多糖具有较强的抗氧化作用。该方法可为黄芪多糖的提取及开发应用提供参考。     

宁波苦丁有效成分提取方法的比较及其抗氧化活性的研究

采用水蒸气蒸馏法(SDE)、乙醇法(EE)、超声波法(UWE)、微波法(MAE)和超临界CO2萃取法(SCDE)提取宁波苦丁有效成分,得率分别为:0.048%、0.011%、0.631%、0.522%和0.935%,毛细管GC-MS法分别鉴定出10、9、12、9和20种成分,体外抗氧化活性测试均显示了抗氧化结果,活性强度依次为:超临界CO2萃取法＞超声波法＞水蒸气蒸馏法＞乙醇法＞微波法.     

抗菌肽对抗生素耐药菌株的抑菌活性研究

[目的]研究抗菌肽对抗生素耐药菌株的抑菌活性。[方法]利用抗性平板划线法从腹泻病牛血便中筛选分离出1株耐药菌,通过16S rDNA序列进行鉴定,采用琼脂孔穴扩散法通过梯度盐酸壮观霉素（spectinomycin,Spe⁺）、氨苄青霉素（ampicillin,Amp⁺）、硫酸卡那霉素（kanamycin,Kan⁺）和氯霉素（chloramphenicol,Cm⁺）试验确定该菌药敏特性,并利用1种抗菌肽制剂对该菌株进行药敏试验。[结果]经BLAST比对分析该菌16S rDNA序列,鉴定该耐药菌为科氏葡萄球菌（Staphylococcus cohnii）,此菌对Amp⁺敏感,但对试验中其他抗生素均有耐药性,各梯度抗菌肽对该耐药菌均具有明显的抑菌活性。[结论]抗菌肽能有效抑制耐药科氏葡萄球菌的生长,有望在畜牧生产中代替抗生素使用。     

产enterocin E5细菌素粪肠球菌发酵条件的优化

采用琼脂扩散法测定粪肠球菌发酵上清液对大肠埃希菌K88的抑菌活性,从而确定产enterocin E5细菌素粪肠球菌的发酵条件。结果表明,粪肠球菌E5产细菌素的发酵培养基为MRS培养基,最适温度为37℃,最适起始pH值6.5,最佳接种量2%,种龄14 h,发酵时间16 h,最佳培养基组分氮源为1%胰蛋白胨、0.5%酵母浸粉,最佳碳源为1%葡萄糖、0.5%蔗糖,0.1%Tween-80有利于enterocin E5的产生。这是分离自北京优良商品猪黑六产enterocin E5粪肠球菌的首次报道。     

Taxonomic and Bioactivity Characterizations of Mameliella alba Strain LZ-28 Isolated from Highly Toxic Marine Dinoflagellate Alexandrium catenella LZT09

Microalgae host varied microbial consortium harboring cross-kingdom interactions with fundamental ecological significance in aquatic ecosystems. Revealing the complex biofunctions of the cultivable bacteria of phycosphere microbiota is one vital basis for deeply understanding the mechanisms governing these dynamic associations. In this study, a new light-yellow pigmented bacterial strain LZ-28 was isolated from the highly-toxic and harmful algal bloom-forming dinoflagellate Alexandrium catenella LZT09. Collective phenotypic and genotypic profiles were obtained to confidently identify this strain as a new Mameliella alba member. Comparative genomic analysis showed that strain LZ-28 shared highly similar functional features with other four marine algae-derived M. alba strains in spite of their distinctive isolation sources. Based on the bioactivity assaying, the mutual growth-promoting effects between bacterial strain LZ-28 and algal strain LZT09 were observed. After the culture conditions were optimized, strain LZ-28 demonstrated an extraordinary production ability for its bioflocculanting exopolysaccharides (EPS). Moreover, the portions of two monosaccharides glucose and fucose of the EPS were found to positively contribute to the bioflocculanting capacity. Therefore, the present study sheds light on the similar genomic features among the selected M. alba strains, and it also reveals the potential pharmaceutical, environmental and biotechnological implications of active EPS produced by this new Mameliella alba strain LZ-28 recovered from toxic bloom-forming marine dinoflagellate.     

饲料中添加蛋白酶AG对鲤鱼鱼种生长和蛋白质消化酶活性的影响

通过2个试验研究了饲料中添加蛋白酶AG对鲤鱼生长和前肠蛋白质消化酶活性的影响.试验Ⅰ选用540尾均重11.7 g的鲤鱼,随机均分为6组,随机选取3组分别饲喂鱼粉含量为10%、15%、20%的3种等蛋白基础饲料,另外3组分别饲喂添加有175 mg/kg蛋白酶AG的以上3种饲料,试验期60 d.试验Ⅱ选取120尾均重48.7 g的鲤鱼,随机分为2组,一组饲喂鱼粉含量6%的基础饲料为对照组,另外一组饲喂添加175 mg/kg蛋白酶AG的基础饲料为试验组,试验期为30 d.试验Ⅰ结果表明:摄食10%鱼粉饲料和20%鱼粉 175 mg/kg蛋白酶AG饲料的鲤鱼分别具有最低和最高的增重率;在10%鱼粉饲料中添加蛋白酶AG显著提高了鱼体增重率(P<0.05),但在15%、20%鱼粉饲料中添加蛋白酶AG,对鱼体增重率没有显著影响.试验Ⅱ结果表明:相比于对照组,试验组鲤鱼增重率提高了6.4%(P<0.05),饲料系数降低了5.4%(P<0.05).试验Ⅰ和试验Ⅱ中,添加蛋白酶AG对鱼体肌肉水分、粗脂肪、粗蛋白质含量均无显著影响(P>0.05);对消化酶活性的测定表明,在鱼粉含量为10%、6%的基础饲料中添加蛋白酶AG,可显著提高前肠组织蛋白酶活性和食糜蛋白酶活性(P<0.05),但在鱼粉含量为15%、20%的基础饲料中添加蛋白酶AG,对前肠组织蛋白酶活性和食糜蛋白酶活性没有影响.综上所述,在鱼粉含量较低的饲料中添加蛋白酶AG,可提高鲤鱼消化道蛋白酶活性,改善生长性能.     

Antihypertensive Effects of IGTGIPGIW Peptide Purified from Hippocampus abdominalis: p-eNOS and p-AKT Stimulation in EA.hy926 Cells and Lowering of Blood Pressure in SHR Model

The aim of this study was to assess the potential hypertensive effects of the IGTGIPGIW peptide purified from Hippocampus abdominalis alcalase hydrolysate (HA) for application in the functional food industry. We investigated the antihypertensive effects of IGTGIPGIW in vitro by assessing nitric oxide production in EA.hy926 endothelial cells, which is a major factor affecting vasorelaxation. The potential vasorelaxation effect was evaluated using 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate, a fluorescent stain. IGTGIPGIW significantly increased the expression of endothelial-derived relaxing factors, including endothelial nitric oxide synthase and protein kinase B, in EA.hy926 cells. Furthermore, oral administration of IGTGIPGIW significantly lowered the systolic blood pressure (183.60 ± 1.34 mmHg) and rapidly recovered the diastolic blood pressure (143.50 ± 5.55 mmHg) in the spontaneously hypertensive rat model in vivo. Our results demonstrate the antihypertensive activity of the IGTGIPGIW peptide purified from H. abdominalis and indicate its suitability for application in the functional food industry.     

Fucose-Rich Sulfated Polysaccharides from Two Vietnamese Sea Cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera: Structures and Anticoagulant Activity

Fucosylated chondroitin sulfates (FCSs) FCS-BA and FCS-HS, as well as fucan sulfates (FSs) FS-BA-AT and FS-HS-AT were isolated from the sea cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera, respectively. Purification of the polysaccharides was carried out by anion-exchange chromatography on DEAE-Sephacel column. Structural characterization of polysaccharides was performed in terms of monosaccharide and sulfate content, as well as using a series of non-destructive NMR spectroscopic methods. Both FCSs were shown to contain a chondroitin core [→3)-β-d-GalNAc-(1→4)-β-d-GlcA-(1→]_n bearing sulfated fucosyl branches at O-3 of every GlcA residue in the chain. These fucosyl residues were different in pattern of sulfation: FCS-BA contained Fuc2S4S, Fuc3S4S and Fuc4S at a ratio of 1:8:2, while FCS-HS contained these residues at a ratio of 2:2:1. Polysaccharides differed also in content of GalNAc4S6S and GalNAc4S units, the ratios being 14:1 for FCS-BA and 4:1 for FCS-HS. Both FCSs demonstrated significant anticoagulant activity in clotting time assay and potentiated inhibition of thrombin, but not of factor Xa. FS-BA-AT was shown to be a regular linear polymer of 4-linked α-L-fucopyranose 3-sulfate, the structure being confirmed by NMR spectra of desulfated polysaccharide. In spite of considerable sulfate content, FS-BA-AT was practically devoid of anticoagulant activity. FS-HS-AT cannot be purified completely from contamination of some FCS. Its structure was tentatively represented as a mixture of chains identical with FS-BA-AT and other chains built up of randomly sulfated alternating 4- and 3-linked α-L-fucopyranose residues.     

丛枝菌根真菌对元宝枫生长及其根系形态的影响

【目的】阐明丛枝菌根（arbuscular mycorrhizal,AM）真菌对元宝枫生长状况及根系形态特性的影响,探讨元宝枫根系形态与其生长发育的关系。【方法】在元宝枫幼苗生长基质中接种摩西斗管囊霉（Funneliformis mosseae）和根内根孢囊霉（Rhizophagus intraradices）2种AM真菌,以不接种AM真菌为对照,通过盆栽试验,培养90 d后,测定元宝枫的生长状况、根系活力及根系形态指标,利用单因素方差分析和多重比较方法,分析接菌与不接菌处理之间的差异,采用envfit ()函数、冗余度分析和变差分解分析元宝枫生长状况与其他指标间的关系。【结果】①接种2种AM真菌后,元宝枫株高、地径、茎干质量和根系活力较对照显著提高,根冠比下降。根内根孢囊霉对元宝枫的促生作用比摩西斗管囊霉显著。②与对照相比,接种2种AM真菌后,元宝枫根表面积和根体积显著增大,粗根根长占总根长的比例增加,细根根长占总根长的比例显著降低。根内根孢囊霉对元宝枫根系形态的影响比摩西斗管囊霉显著。③元宝枫生长状况与其根系活力和根系形态密切相关。【结论】接种AM真菌有助于增强元宝枫的根系活力,改善其根系形态,促使其汲取更多养分,进而提高其生物量。     

6个鲜食葡萄品种不同部位生物活性物质含量及抗氧化活性比较

【目的】 对不同色系鲜食葡萄品种各器官生物活性物质含量和各器官提取物总活性进行比较研究,为鲜食葡萄育种和生物活性物质利用提供有效依据。【方法】 以三种色系鲜食葡萄(各含一个有籽和无籽鲜食葡萄品种)为材料,分别测定果梗、果皮、果肉、葡萄籽中多酚、黄酮、原花青素含量,用自由基清除法评价各器官提取物的总活性,并使用相关系数法分析3种生物活性物质含量和总活性的相关性。【结果】 结果表明,不同色系葡萄品种中,黄绿色系与红色系果梗多酚含量高于紫黑色系品种,无籽品种含量高于有籽品种;黄酮含量以红色系品种含量最高,且无籽高于有籽品种。果皮中多酚、黄酮及原花青素含量与果皮颜色深浅有关,颜色越深含量越高;原花青素含量在黄绿色系与红色系葡萄品种中无籽显著高于有籽葡萄。果肉多酚含量紫黑色系有籽葡萄显著低于其它品种;原花青素含量紫黑色系显著高于红色系品种,黄绿色系及红色系品种有籽显著高于无籽品种。葡萄籽中多酚及原花青素含量变化具有随着果皮颜色加深而显著增加的变化趋势。生物活性物质抗氧化活性在果梗中黄绿色系与红色系品种显著高于紫黑色系品种,无籽品种高于有籽品种;在果皮中紫黑色系品种显著高于黄绿色系与红色系品种;在果肉中黄绿色系有籽品种显著高于无籽品种;多酚、黄酮含量与提取物总的抗氧化活性呈极显著正相关关系(P <0.01)。综上可知,鲜食葡萄果梗中含有丰富的多酚和黄酮物质,且红色系无籽品种显著最高,可将葡萄采果后果梗作为多酚和黄酮物质提取的原材料;鲜食葡萄多酚、黄酮、原花青素等生物活性物质随着果皮颜色加深而加深,可选育果皮颜色深且品质好的品种做鲜食葡萄;葡萄籽中含有丰富的多酚、黄酮、原花青素,可作为生物活性物质提取的原材料;鲜食葡萄提取物总抗氧化活性主要由多酚、黄酮决定。