生物传感器在渔业监测中的研究进展

论述了生物传感器在监测渔业环境中的BOD、N、P、病原体、农药和重金属等方面的国内外最新进展 ,分析了生物传感器在海洋渔业应用中存在的主要问题及原因 ,同时阐述了利用基因工程微生物技术、分子烙印技术、多成分实时分析技术等先进技术提高生物传感器的性能并扩大其在海洋渔业领域中的应用范围 ,在海洋渔业环境中为实现生物传感器与计算机在线自动监测与控制 ,开拓新的广阔前景。     

微生物全细胞传感器在重金属生物可利用度监测中的研究进展

传统的物理化学方法主要用于测定环境重金属的总量,微生物全细胞传感器可以对土壤及水体环境的重金属生物可利用度进行监测.此外,微生物全细胞传感器还具有操作简单、快速、经济的特点,适用于污染事件的应急监测.微生物全细胞传感器的生物学元件主要由MerR、ArsR、RS等家族的金属调控蛋白和gfp、lux、luc等报告基因组成.调控蛋白、报告基因与微生物全细胞传感器的灵敏度、特异性和监测特点有关.受pH、金属螯合物及检测条件等因素的影响,不同的环境条件下的重金属生物可利用度是不同的.增加重金属在微生物细胞内的累积,进行调控蛋白的分子生物学改造,优化检测条件是提高传感器灵敏度、特异性和准确性的可行方案.实现污染物的原位和在线监测是微生物全细胞传感器的主要发展方向.     

Overexpression of NADH oxidase gene from Deinococcus geothermalis in Escherichia coli

When using stable enzyme genes from a thermophile to create a biosensor in Escherichia coli, it is vital that these genes be overexpressed in order to provide a sufficient supply of enzymes. In this study, overexpression of the NADH oxidase (Nox) gene from the thermophile Deinococcus geothermalis was successfully achieved with the aim of creating a stable biosensor active at room temperatures. To do so, modification of 10 nucleotides, GAAATTAACT, upstream of the start codon of the Nox gene was necessary.     

Overexpression of NADH oxidase gene from Deinococcus geothermalis in Escherichia coli

When using stable enzyme genes from a thermophile to create a biosensor in Escherichia coli, it is vital that these genes be overexpressed in order to provide a sufficient supply of enzymes. In this study, overexpression of the NADH oxidase (Nox) gene from the thermophile Deinococcus geothermalis was successfully achieved with the aim of creating a stable biosensor active at room temperatures. To do so, modification of 10 nucleotides, GAAATTAACT, upstream of the start codon of the Nox gene was necessary.     

Resistance and biosorption mechanism of silver ions by Bacillus cereus biomass

Biosorption of silver ions onto Bacillus cereus biomass was investigated. Overall kinetic experiments were performed for the determination of the necessary contact time for the attainment of equilibrium. It was found that the overall biosorption process was best described by pseudo second-order kinetic model. The crystals detected by scanning electron microscope and X-ray photoelectron spectroscopy suggested the precipitation was a possible mechanism of biosorption. The molecular genetics of silver resistance of B. cereus biomass was also detected and illustrated by a whole cell sensor tool.     

A GFP-based bacterial biosensor with chromosomally integrated sensing cassette for quantitative detection of Hg(II) in environment

A mercury biosensor was constructed by integrating biosensor genetic elements into E. coli JM109 chromosome in a single copy number, using the attP/attB recombination mechanism of λphage. The genetic elements used include a regulatory protein gene (merR) along with operator/promoter (O/P) derived from the mercury resistance operon from pDU1358 plasmid of Serratia marcescens. The expression of reporter gene gfp is also controlled by merR/O/P. Integration of the construct into the chromosome was done to increase the stability and precision of the biosensor. This biosensor could detect Hg(II) ions in the concentration range of 100-1700 nmol/L, and manifest the result as the expression of GFP. The GFP expression was significantly different (P ≤ 0.05) for each concentration of inducing Hg(II) ions in the detection range, which reduces the chances of misinterpretation of results. A model using regression method was also derived for the quantification of the concentration of Hg(II) in water samples.     

基于平面波导型荧光免疫传感器的双酚A检测适用性研究

双酚A是一种环境内分泌干扰物,逐渐受到国内外广泛关注.平面波导型荧光免疫传感器可以对水样中存在的痕量双酚A进行快速高灵敏度检测.在最优检测条件下测得双酚A标准曲线的检出限为(0.04±0.007)μg·L-1,线性区间为0.16~22.40μg·L-1,半抑制浓度为(1.67±0.47)μg·L-1.加入0.5%的EDTA到样品溶液中可以削弱水体硬度的干扰,并在最优条件下测得4种实际水样的加标回收率在88%~111%之间,相对标准偏差小于15%,表明该方法可以运用于实际水样中双酚A的检测.     

单室MFC型生物毒性传感器对重金属离子的检测研究

构建了单室微生物燃料电池(air-cathode microbial fuel cell,ACMFC)型生物毒性传感器,以含重金属离子(Cd2+、Cu2+)人工配水为检测对象,分析了有毒物质对检测仪的抑制率与有毒物质浓度的线性关系.结果表明,①单室MFC型生物毒性传感器结构简单,操作方便,灵敏度较高,可用于水体重金属离子(Cd2+、Cu2+)生物毒性的快速检测;②在实验条件下,该生物毒性传感器检测时间4h,清洗时间2～10min,恢复时间4h;③检测结果显示,实验室自配Cd2+、Cu2+及其混合液IC20值分别为0.6、0.8和0.25mg/L,有毒物质浓度与MFC产电量的抑制率呈显著正相关,相关系数分别为0.9960、0.9744和0.9907.     

微生物传感器测定河豚毒素研究

以地衣芽孢杆菌、假单胞菌和枯草芽孢杆菌为识别元件,采用夹层法固定化微生物膜与氧电极组成毒性微生物传感器,实验了3种不同菌株生物传感器对河豚毒素的响应能力,得出对河豚毒素敏感的微生物菌株为地衣芽孢杆菌.实验得出地衣芽孢杆菌传感器最佳工作条件为pH=7.76,温度35℃,底物GGA质量浓度为21.5 mg/L.在此条件下传感器对河豚毒素响应的标准曲线y=0.0025x+0.0122,线性范围为10～100 μg/mL,相关系数0.9776.     

苯酚在预活化聚酰胺膜酪氨酸酶生物传感器上的响应特性

研究苯酚在预活化聚酰胺膜酪氨酸酶生物传感器上的响应特性,适量儿茶酚的存在可加快传感器到达恒态的速度。被固定酶的量、活性及传感器的有效面积等是影响检测灵敏度的主要因素。首次采用预活化聚酰胺膜固定酪氨酸酶,并与浸蜡石墨电极组合成酪氨酸酶生物传感器,在温度２５℃电位－０．２００ＶｖｓＳＣＥ,儿茶酚浓度５×１０（－７）ｍｏｌ／Ｌ,底液０．１ｍｏｌ／Ｌ磷酸盐缓冲溶液ｐＨ６．５０下,采用电流法测定苯酚及水样中的酚。酶膜的制备、替换和贮存极为方便,酶膜贮存５个月未见活性下降,有良好的重现性和线性范围２×１０（－７）－１．２５×１０（－５）ｍｏｌ／Ｌ。