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Cloning and expression of the first gene for biodegrading microcystin LR by Sphingopyxis sp. USTB-05
Hai Yan Huasheng Wang Junfeng Wang Chunhua Yin Song M Xiaolu Liu Xueyao Yin 《环境科学学报(英文版)》2012,24(10):1816-1822
Harmful cyanobacterial blooms are a growing environmental problem worldwide in natural waters, the biodegradation is found to be the most efficient method for removing microcystins (MCs) produced by harmful cyanobacteria. Based on the isolation of a promising bacterial strain of Sphingopyxis sp. USTB-05 for biodegrading MCs, we for the first time cloned and expressed a gene USTB-05-A (HM245411) that is responsible for the first step in the biodegradation of microcystin LR (MC-LR) in E. coli DH5αup, with a cloning vector of pGEM-T easy and an expression vector of pGEX-4T-1, respectively. The cell-free extracts (CE) of recombinant E. coli DH5αup containing USTB-05-A had high activity for biodegrading MC-LR. The initial MC-LR concentration of 40 mg/L was completely biodegraded within 1 hr in the presence of CE with a protein concentration of 0.35 mg/mL. Based on an analysis of the liquid chromatogram-mass spectrum (LC-MS), the enzyme encoded by gene USTB-05-A was found to be active in cleaving the target peptide bond between 3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-deca-4,6-dienoic acid (Adda) and arginine of MC-LR, and converting cyclic MC-LR to linear MC-LR as a first product that is much less toxic than parent MC-LR, which offered direct evidence for the first step on the pathway of MC-LR biodegradation by Sphingopyxis sp. USTB-05. 相似文献
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He Zhenrong He Jiawan Yu Minjuan Qiao Mingye Shen Zhi Wu Liandi Wu Suozhu Guo Yunfeng Fumio Kondo Ken-ichi Harada 《环境科学学报(英文版)》1997,9(1):113-119
Seven types of microcystins,isolated from Microcystis waterbloom in Lake Dalai, were characterized.The major toxins:MCYST-LR,MCYST-RR,[D-Asp3]MCYST-LR and [Dha7]MCYST-LR were identified by high performance liquid chromatography(HPLC),as compared with the authentic microcystins.The minor toxins:MCYST FR,[L-Mser7]MCYST-LR and an unknown MCYST which was most likely to be MCYST-(H4)YR were identified with fritfast atom bombardment liquid chromatography/mass spectrometry(Frit-FAB LC/MS)and amino acid analysis.The toxigenic diversity in blue-green algae(cyanobacteria)was discussed. 相似文献
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微囊藻毒素在土壤中的环境行为及污染风险 总被引:1,自引:0,他引:1
微囊藻毒素(Microcystins,MCs)是富营养化水体中发生蓝藻水华时所产生的一类肝毒素,在环境中大量出现时将对生态系统带来冲击并可通过食物链进入人体进而危害人类健康.关于微囊藻毒素的环境行为和污染风险近年来已成为研究热点.论文基于地表物质循环原理,分析了水体中微囊藻毒素经土壤进入人体的途径,指出了土壤在微囊藻毒素迁移转化链条中的位置,概述了微囊藻毒素在土壤中的环境化学行为、农作物的吸收特性及其生态毒理效应等方面的研究进展,提出了在微囊藻毒素迁移转化过程中土壤的净化和传递两大功能,并在此基础上展望了今后的研究方向. 相似文献
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珠江三角洲城市周边典型中小型水库富营养化与蓝藻种群动态 总被引:8,自引:0,他引:8
为了解珠江三角洲地区城市周边中小型水库的水质状况,于2009年8月(丰水期)和2010年3月(枯水期)调查了4座典型中小型水库--横岗水库、水濂山水库、契爷石水库和东风水库,分析了水库的富营养化与蓝藻种类组成、种群动态及微囊藻毒素水平.丰水期总磷浓度为0.05~0.083 mgL-1,枯水期为0.026~0.082 m... 相似文献
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水华期间微囊藻毒素的释放会严重影响洱海饮用水源水质安全.应用荧光定量PCR结合酶联免疫法快速检测洱海水华高发秋季(8—11月)主要旅游区和饮用水源地产毒微囊藻丰度和微囊藻毒素LR浓度.结果表明,荧光定量PCR方法能够快速定量洱海中总微囊藻种群和产毒微囊藻丰度,并且能有效预测洱海水体产毒潜能.洱海总微囊藻平均丰度为8.15×10~6copies·L~(-1),9月份均值最高,产毒微囊藻平均丰度为6.42×10~5copies·L~(-1),也于9月达到最高值,占总微囊藻的比例为1.0%~69.8%,水温和TP显著影响洱海总微囊藻丰度,低温、低P是洱海微囊藻水华的限制因子.洱海毒素峰值期为10月上旬,期间水源地微囊藻毒素和胞外微囊藻毒素全部检出,叶绿素a显著影响总微囊藻毒素和胞外微囊藻毒素分布,说明在一定程度上Chl a值能预测水体微囊藻释放的毒素风险.洱海总微囊藻毒素LR最高值达2.17μg·L~(-1),已超过集中式生活饮用水地表水源地对MC-LR的限值(1.0μg·L~(-1)),表明水华期间洱海饮用水水源安全问题不容忽视. 相似文献
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以野外收集的水华蓝藻为原料,经过75%甲醇溶液浸提,快速色谱分离和半制备色谱纯化,从滇池水华蓝藻中分离纯化出1种微囊藻毒素变体。电喷雾质谱、紫外分光光度计和HPLC检测结果表明,所得毒素为[Dha7]MCRR,是MCRR的1种去甲基化变体,其纯度大于 95%。该毒素的分子组成为环(Ala-Arg-MeAsp-Arg-Adda-Glu-Dha),分子量为1023,其紫外扫描光谱(200~300nm)在239nm处有特征吸收。[Dha7]MCRR在滇池水华蓝藻中普遍存在,有时会成为MC的主要种类。 相似文献
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铜绿微囊藻及其所产生的藻毒素严重威胁人类健康和生命安全,给水处理带来很大困难.利用直线加速器产生的电子束对铜绿微囊藻分别进行1、2、3、4和5 kGy的辐射,通过胞内毒素和胞外毒素的变化,分析了电子束辐射对藻细胞毒素产生和释放过程的影响.结果表明,2~5 kGy的辐射使藻细胞叶绿素a含量迅速大幅下降,对胞外毒素的去除率分别为76.9%、76.2%、76.2%和97.7%,表明电子束辐射对微囊藻毒素的降解作用.胞内毒素、胞外毒素与叶绿素a浓度之间的相关性拟合进一步证实电子束辐射抑制了胞内毒素的产生. 相似文献