Joint Toxicological Effects and Proteomics in Zebrafish Brain under the Combination Stress of Methyl Parathion with Cadmium

对多种污染物产生联合毒性效应的研究已成为环境科学发展的重要方向之一。目前，有关污染物的生物效应研究多数还是集中在单个化合物，因而有关预测和治理多种毒物对生物产生联合毒性的研究，越来越受到环境科学家的关注和重视，特别是有关有机磷农药和重金属的联合毒性效应研究。本文选取了在水体污染中比较有代表性的甲基对硫磷(MP)和重金属镉(Cd)为共污染物，以经典的模式生物斑马鱼(Daniorerio)为实验材料，从生理生化指标的变化、蛋白的差异表达等几方面对MP和Cd对斑马鱼脑组织的联合胁迫效应进行了较为系统的研究。 首先，对MP和Cd分别胁迫斑马鱼后产生的急性毒性效应进行分析。结果显示MP对斑马鱼的半致死...Combined pollution has become one of the important directions of research in environment science at present. Before now most of the research has concentrated on the action of single compound against organisms. Due to the presence of multiple pollutants in natural environments, environmental scientists are paying more attentions on the joint toxicological effects, especially about the combination o...学位：理学博士院系专业：化学化工学院化学系_化学生物学专业学号：1912005140309

一种富集高温栅藻Desmodesmus sp. F51的新型生物絮凝剂

【背景】海科贝特氏菌(Cobetia marina)可产生大量具有絮凝活性的胞外产物，可视为一种新型的生物絮凝剂。高温栅藻(Desmodesmus sp. F51)是一种具有较高叶黄素含量的微藻，被认为是一种新兴的叶黄素来源，但利用该生物絮凝剂高效富集高温栅藻的相关研究迄今尚未见报道。【目的】以高温栅藻为对象，研究该新型生物絮凝剂的絮凝效率，并对絮凝机理进行初步探讨。【方法】探索在不同生长阶段微藻培养液添加生物絮凝剂、添加量、絮凝时间、pH对絮凝效率的影响，分析生物絮凝剂的功能基团，并测定在不同pH条件下添加生物絮凝剂前后高温栅藻的Zeta电位变化，以及在显微镜下分析藻细胞在添加生物絮凝剂前后的形态。【结果】在高温栅藻生长至稳定期，pH 8.0添加2 mL生物絮凝剂，絮凝15 min絮凝效果最佳，达82.1%。傅里叶红外图谱显示了多糖及酰胺结构的特征吸收峰，由此推测生物絮凝剂主要是多糖的混合物，含有少量蛋白质。根据Bradford法测定絮凝剂中蛋白含量约为0.4%(质量比)，通过苯酚-硫酸法测定总糖质量分数约为34.5%(质量比)，与FTIR谱分析结果基本相符。生物絮凝剂在pH 4.0～11.0保持60%以上的絮凝效率，说明无论是酸性或是碱性条件下絮凝效率都较高，结合Zeta电位的分析表明，推测生物絮凝剂对高温栅藻的絮凝机理中占主导地位的可能是吸附架桥作用。【结论】该研究对微藻生物絮凝具有重要的理论和实践意义。厦门市科技计划项目(3502Z20173018)福建省自然科学基金(2017J01077)国家自然科学基金项目 (31871779

Domestication of the high-sugar-tolerant Mortierella alpina on arachidonic acid(ARA) production

通讯作者：Tel: 86-592-2186038; : [email protected][中文摘要]【目的】提高花生四烯酸(Arachidonic acid,ARA)产量,克服ARA产生菌高山被孢霉(Mortierella alpina)在长期的保存及使用过程中易受到外界条件影响发生退化,从而导致菌种耗糖量降低、影响菌种摄入营养的能力和不利于工业化生产的缺点。【方法】首先采用固体培养基驯化,将菌种逐级涂布于梯度高糖PDA平板(含糖量分别为2%、5%、7%、10%和15%)培养,挑选经固体驯化后能耐受10%高糖浓度平板的菌种,转接到两种含不同氮源的梯度高糖(含糖量分别为3%、4%、5%和6%)液体培养基中进行驯化,最后对驯化后的菌种进行2 L发酵罐放大实验。【结果】当培养基中以酵母粉为氮源时,驯化后菌体的最高耗糖量由3 g/(L.d)提高到12 g/(L.d);当培养基中以玉米浆为氮源时,驯化后菌体的最高耗糖量由7 g/(L.d)提高到12 g/(L.d)。摇瓶驯化实验结果表明以玉米浆为氮源驯化的菌种发酵效果较好,发酵罐实验结果显示菌体生物量为50 g/L,总油脂为18 g/L,目的产物ARA产量为8 g/L。相比未驯化之前的发酵结果,生物量和总油脂含量提高了近3倍,ARA产量提高了近4倍。【结论】经过高糖驯化, 菌种的耗糖能力得到提高, 生物量、总油脂及ARA 的产量也都有所增加, 从而可以使菌种在保存和使用过程中不易退化,保持稳定。[英文摘要][Objective] In order to improve arachidonia acid(ARA) production,and prevent the degeneration of ARA-producing strain Mortierella alpina in long-term culture preservation and cultivation,which could lead to low consumption rate of substrates like carbon source.[Methods] Mortierella alpina strain was first domesticated in high-sugar PDA plate with gra-dient sugar content(2%、5%、7%、10% and 15%).The strain which grew better in the solid medium containing 10% sugar was then selected and transferred to two liquid high-sugar media with different nitrogen sources to domesticate. The gradient sugar contents were 3%、4%、5% and 6%, respectivlely. Finally, the domesticated strain was used to produce ARA in a 2 L bioreactor. [Results] The experimental results show that the consumption of sugar increased from 3 g/(L·d) up to 12 g/(L·d) in the medium with yeast extract, and from 7 g/(L·d) up to 12 g/(L·d) in the medium with corn steep liquor. After cultivation for 144 h in a 2 L bioreactor,the yield of ARA reached 8 g/L, which was 4 times higher than that obtained with the original strain. [Conclusion] The domestication of the high-sugar-tolerant Mortierella alpina increased the consumption of sugar, biomass, total fatty acid and ARA production. The domesticated strain could maintain the high sugar resistance in the preserving process and turn to be more stable.福建省科技厅重大专项项目(No.2010NZ0001-4);; 国家自然科学基金项目(No.31071488

Preparation of Supported Palladium Catalyst by Bioreduction

［中文文摘］将微生物可在常温下还原贵金属离子的特性引入催化剂的制备过程中,利用对Pd2+具有较强还原能力的地衣芽孢杆菌(简称R08)制得负载型Pd催化剂(简称催化剂)。采用X射线光电子能谱(XPS)和透射电镜(TEM)对催化剂进行表征。XPS测定结果表明,室温下R08菌体可将γ-A l2O3载体表面上的Pd2+基本还原为Pd0;生物还原法制得的催化剂的Pd微粒的平均粒径约为5nm。将该催化剂用于2%CO-98%空气(体积分数)混合气的催化氧化反应,CO完全氧化的最低反应温度为60℃,在此温度下催化剂的活性可恒定150h,结果优于相同条件下化学浸渍法制得的催化剂。XPS表征和催化活性评价结果说明,用于CO催化氧化反应的催化剂中单原子Pd活性中心的价态为0~+2。［英文文摘］Pd catalyst supported on γ2a lumina ( nanoparticles) with high dispersion was prepared by bioreduction with Bacillus lichenifoum is ( strain R08) , which was strong in reducing ab ility of Pd2 + . Catalysts prepared by bioreduction and impregnation were cha racte rized by m eans of XPS and TEM.XPS spectra indicated tha t s train R08 could almost completely reduce Pd2 + to Pd0. TEM images showed that average sizes of Pd particles on catalysts prepared by bioreduction and by impregnation were 5 nm and 18 nm, respectively. When catalyst p rep ared by bioreduc tion w as used in ox idization of carbon m onoxide to carbon dioxide, the carbon monoxide could be comp letely oxidized at the lowest temperature of 60 ℃and activity of catalyst could be main tained a t this tempe rature fo r 150 h. The result was better than catalyst prepared by imp regnation. Pd ( Ph2 PCH2 PPh2 ) 2 suppo rted on γ2alum ina was synthesized as catalyst for the oxidation. Results of XPS spectra and activity eva luation indicated that chemical valence state of Pd on active center of catalyst was between 0 and + 2.国家自然科学基金项目(20376067

Expression of α-1,6-dextranase from Penicillium minioluteum in P.pastoris

α-1,6-葡聚糖酶是专一作用于α-1,6糖苷键产生小分子葡聚糖的一类水解酶,广泛的运用于制糖工业和啤酒工业中。采用PCr法扩增朱黄青霉(PEnICIllIuM MInIOluTEuM)C12114的α-1,6-葡聚糖酶基因,将其插入毕赤酵母表达载体PPIC9k。经SACI酶线性化电击转入毕赤酵母基因组,构建重组酵母gS115/PPIC9k-dEX。对构建成功的转化子进行1.5%的甲醇诱导表达,在30℃条件下培养7d时酶活达到最大值,为88.35u/Ml。α-1,6-dextranase,which can hydrolyze dextran specifically by cutting off the α-1,6-glycosidic bond to release shorter saccharides,was widely used in many fields such as sugar industry and beer industry.The gene of α-1,6-dextranase(dex)was amplified through PCR by using Penicillium minioluteum C12114 genomic DNA as template.The amplified gene was cloned into vector pPIC9K and the recombinant plasmid pPIC9K-dex was linearzed with Sac I,then transformed into P.pastoris GS115 by electroporation.The positive transformant was induced to express the enzyme with 1.5% methanol for 7 days under the 30℃,and the activity of the enzyme could reach 88.35U/mL.广东省科技计划项

Studies on mechanisms of silver biosorption by Aeromonas SH10

［中文文摘］以气单胞菌SH10为生物吸附材料,从静电吸附作用、离子交换作用、络合作用、沉淀作用几方面研究了SH10吸附Ag+过程可能存在的作用机制.结果表明,气单胞菌SH10吸附银离子依赖于静电吸附作用;吸附Ag+前后溶液中Na+、K+、Mg2+浓度的变化较小,说明吸附过程中离子交换不明显;SH10表面的羧基、氨基和脂类基团经化学屏蔽后,对Ag+的吸附量明显下降,结合红外光谱分析结果,确定酰胺基CO-NH为SH10与银离子发生络合作用的主要官能团;电镜扫描结果表明,SH10在吸附过程中可以和Ag+形成沉淀沉积在细胞表面.［英文文摘］The study on mechanisms of metal biosorption is helpful to improve the biosorption ability of organism. Mechanisms of silver biosorption by Aeromonas SH10 were discussed from several aspects such as electrostatic adsorption, ion exchange, complexation and deposition. Results showed that silver biosorption by SH10 didn't mainly depend on ion exchange duo to only slight increase of the contents of Na~+, K~+, Mg~ 2+ in solution after biosorption, while electrostatic adsorption was found to be responsible for biosorption. Adsorptive capacity of silver ion decreased distinctly after functional groups such as carboxyl, amino and lipid groups on SH10 cell wall were chemicallymodified, respectively. Combined with FTIR spectra, amino groups were confirmed to be the main active groups that could combine with silver ion. SEM illustrated precipitation could take place on the cellwall of SH10.国家自然科学基金(No.20376067

Study on mechanisms of silver biosorption by laminaria japonica

生物吸附金属作用机制的研究,有助于提高生物吸附剂的吸附能力。文章以前期优选出的对银具有较强吸附能力的海带为生物吸附材料,从静电吸附作用、离子交换作用、络合作用、沉淀作用几方面研究了其吸附Ag+过程可能存在的作用机制。结果表明:海带吸附银离子不依赖于静电吸附作用;吸附Ag+前后溶液中Na+、K+、Mg2+质量浓度的变化说明吸附过程中存在离子交换机制;海带表面的羧基、氨基和脂类基团经化学屏蔽后,对Ag+的吸附量明显下降,结合红外光谱分析结果,确定酰胺基CO—NH和离子化羧基COO-为海带与银离子发生络合作用的主要官能团;电镜结果表明海带在吸附过程中可以和Ag+形成沉淀沉积在细胞表面。The study on mechanisms of metal biosorption is helpful to improve the biosorption ability of organism.Mechanisms of silver biosorption by Laminaria japonica were discussed from several aspects such as electrostatic(adsorption,) ion exchange,complexation and deposition.The results show that silver biosorption by Laminaria(japonica) doesn′t depend on electrostatic adsorption,while ion exchange is responsible for biosorption because of(increase) of the mass concentration of Na~+,K~+,Mg~(2+) in solution before and after biosorption.Adsorptive capacity of Ag~+ distinctly decreases after functional groups such as carboxyl,amino and lipid groups in Laminaria japonica cell wall are chemically modified respectively.Combined with FTIR spectra,carboxyl and amino groups are(confirmed) to be the main active groups which can combine with silver ion.Precipitation can take place on the cell wall of Laminaria japonica through SEM characterization.国家自然科学基金资助项目(20376067

Hot water extraction of longan polysaccharide assisted by microwave pretreatment

联系人: 李清彪。第一作者: 杨翠娴( 1982 ) ) , 女, 硕士研究生。龙眼多糖具有抗氧化、抗衰老等多种生物活性,但对龙眼多糖提取工艺和其化学结构方面的研究文献报道较少。采用微波前处理-热水浸提新工艺提取龙眼多糖,单因素考察及正交实验结果表明,在所考察实验范围内,龙眼多糖的最佳提取工艺条件为:微波前处理功率700W,处理时间60s,热水浸提料液比1∶15,浸提温度100℃,浸提时间7h,搅拌速率240r.min-1。在此条件下,龙眼多糖收率可达9.00mg.(g龙眼)-1(干重)。紫外和红外光谱分析结果显示,所获得的龙眼多糖是具乙酰氨基结构的β型吡喃酸性杂多糖。厦门市翔安区科技局资助项

优化提取与分离海兔大脑神经节蛋白质组

蓝斑背肛海兔(Notarcus Leachii cirrosusStimpson,NLCS)属于海洋软体动物.NLCS大脑神经节(cerebral ganglion,CG)富含高丰度的脂类物质,并干扰了三氯醋酸-丙酮法沉淀CG蛋白质.本实验的目的是找到一种适合提取、分离海兔CG蛋白的方法.经各种实验方法对比后,发现超速离心法能有效分离CG破碎液中的脂类化合物,提高双向凝胶电泳法分离BG蛋白质组的分辨率,大约可检测到450个蛋白质点

Secretive Expression and Optimized Inducement of Barnase

从解淀粉芽孢杆菌中PCR分别扩增解淀粉芽孢杆菌核糖核酸酶barnase基因及其抑制剂barstar基因,采用将barnase基因置于barstar基因保护下的克隆策略,以pET-22b(+)质粒为基础,构建大肠杆菌分泌型表达质粒.IPTG诱导表达目的蛋白后将培养基蛋白进行SDS-PAGE分析并从诱导温度、IPTG诱导浓度和诱导时间三方面初步优化诱导表达条件.Using PCR method to amplify Bacillus amyloliquefaciens ribonuclease barnase gene and its inhibitor barstar gene from Bacillus amyloliquefaciens. The secretive expression vector is constructed by the strategy of protective clone from plasmid pET-22b(+). After induced by IPTG, a 12 kD target barnase protein can be detected from medium protein by SDS-PAGE analysis. Inducing conditions of temperature, IPTG concentration and inducing time have also been optimized.福建省高等学校新世纪优秀人才支持计划资助项