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The activities and substrate specificities of extracellular enzymes in natural systems are not well understood, despite their critical role in microbial remineralization of organic carbon. These enzymes initiate organic carbon degradation by selectively hydrolyzing high molecular weight substrates to lower molecular weight products which can be transported into cells. A set of single- and dual-labeled fluorescent polysaccharides was synthesized and characterized to explore a variety of approaches for measuring enzymatic hydrolysis of biopolymers via photophysical techniques, focusing particularly on rapid and robust optical techniques which are amenable to field measurements in remote locales. A shotgun-labeling approach yielded dual-labeled probes that exhibited substantial donor fluorophore quenching. The photophysical response of these probes to hydrolysis via purified enzymes was investigated in the lab, and fluorescence polarization proved to be a rapid and reliable technique for monitoring probe hydrolysis. Initial field results were also obtained from hydrolysis experiments in sediment porewaters. Because polarization measurements are rapid and simple, this approach could be used to follow the extracellular enzymatic hydrolysis of a wide range of biopolymers which fuel microbial metabolism. 相似文献
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Effects of heavy metals on some soil biological parameters 总被引:11,自引:0,他引:11
Olga Mikanova 《Journal of Geochemical Exploration》2006,88(1-3):220
The soils most polluted by heavy metals are probably alluvial soils. One of them is the alluvium of the Litavka River. The heavy metal content was much higher in some cases than the threshold values, allowed by the Czech standards. The effect of soil contamination on enzyme activity is presented. Generally, the values of enzymatic activities were highest in the soil above the source of contamination and they decrease as they approach the source of contamination. All measured parameters were affected by heavy metal concentrations. Our results demonstrate that several parameters of microbial activity could be used as good indicators of increasing concentrations of Cd, Pb and Zn in soil. 相似文献
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以海洋脂肪酶ADM47601为研究对象,以树脂为载体,进行了固定化酶的研究。筛选了21种树脂,确定D316型阴离子交换树脂为载体。通过正交试验得到了固定化的最佳条件,每克载体加脂肪酶10 000 U,pH7.5,25℃,150 r/min,固定化时间3 h,固定化酶活力回收率可达65.53%±1.06%。固定化脂肪酶最适作用pH为7.5,在pH6.0~9.0的范围内能够保持85%的活力。最适作用温度40℃,在50℃下贮存1 h后依然保持66%的活力。25℃保存半衰期为55 d,操作8次仍然能维持60%以上的活性,且在多种有机试剂中能够稳定的保持活性。固定化脂肪酶Km和Vmax值分别为4.86×10~(–5) mol/L和1.31×10~(–5) mol/(L·min),对底物亲和性较好。 相似文献
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During spring the atmospheric concentration of Peroxyacetyl Nitrate, PAN, in the UK varies over three orders of magnitude due to long range transport and local photochemical production. An experiment at Great Dun Fell was used to determine which of these sources of PAN controlled spring time concentrations and to quantify the contribution of PAN to oxidised nitrogen in cloud water. The gas phase mixing ratio of a wide range of trace gas pollutants, including PAN, along with cloud water chemical composition and aerosol/cloud droplet distributions were measured. A chemical model was developed to determine the contribution of PAN hydrolysis to cloud water nitrification and to quantify the sink strengths of PAN. Chemical reaction following thermal decomposition was found to be the dominant sink mechanism in the model but transport was the most important factor controlling PAN concentration. During cloud episodes PAN concentrations varied from 100 ppt to over 1 ppb but the calculated inorganic oxidised nitrogen loading in the cloud due to PAN dissolution and hydrolysis was an insignificant fraction of the observed total oxidised nitrogen loading of the cloud. Extrapolation of the model to persistent cloud in marine conditions showed that PAN has a negligible effect on cloud nitrification. It was concluded that PAN concentrations were heavily influenced by regional concentrations and that the hydrolysis of PAN in cloud water was an insignificant source of oxidised nitrogen in cloud water. 相似文献
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采用生物蛋白酶酶解罗非鱼鱼排蛋白,以水解度(DH)为指标,进行了单酶、复合酶酶解效果的比较分析,并利用响应面分析方法(RSM)对酶解参数进行优化。结果表明,采用风味蛋白酶和木瓜蛋白酶双酶组合分段酶解,固定总酶添加量为3.0%、双酶复合比2:1、酶解时间3h、分段酶解时间比0.6:1,酶解温度56℃,分段酶解pH为7.0和6.2时,酶解效果最佳,罗非鱼鱼排蛋白水解度达到32.49%,酶解产物中游离氨基酸含量达32.84mg/g鱼排,占罗非鱼鱼排总氨基酸含量的25.43%,其中呈味氨基酸含量达18.48%,必需氨基酸含量达67.96%。 相似文献
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以鳕鱼皮为原料,采用双酶水解法制备鱼蛋白水解物再经亚铁修饰获得鱼蛋白水解物亚铁复合物(Fe-FPH),并对Fe-FPH进行了抗营养性贫血的实验研究。结果表明:(1)以水解度(DH)为5%的鱼蛋白水解物为原料,在pH7.0、温度20℃、螯合时间15min的条件下可获得Fe-FPH6.7%;(2)Fe-FPH蛋白含量为85.1%,铁含量达40254mg/kg,氨基酸价为59.62;(3)抗营养性动物实验研究表明:Fe-FPH给药组第5天可显著提高营养性贫血大鼠的Hb含量,第11天时表现出极显著效果;Fe-FPH组,第11天有显著降低大鼠血中PPIX作用,到第18天时表现出极显著效果。因此,Fe-FPH不仅含有较高的营养价值,而且有显著抗营养性贫血功能,可望成为鳕鱼加工副产物高效利用的有效途径。 相似文献