狐尾藻化感物质的提取工艺优化及其急性毒性研究

发布时间：2018-05-17 05:23

  本文选题：铜绿微囊藻 + 化感物质　； 参考：《上海海洋大学》2015年硕士论文

【摘要】：近年来,环境污染日趋严重,水体富营养化不断加剧,水华频繁,铜绿微囊藻(Microcystis aeruginosa)作为常见水华蓝藻,给人们生活和工农业发展带来严重威胁。本研究以狐尾藻(Myriophyllum spicatum)茎部化感物质为研究对象,分析超声波-纤维素酶法提取狐尾藻化感物质的最佳工艺条件,探讨提取液对铜绿微囊藻细胞抗氧化酶系统的影响以及斑马鱼的急性毒性。研究发现超声波-纤维素酶提取液能快速有效的抑制铜绿微囊藻的生长,破坏藻细胞抗氧化酶系统,杀死藻细胞,但是对斑马鱼为低毒性物质,为今后进一步的研究提供一定的理论支持。采用超声波-纤维素酶法提取狐尾藻的抑藻成分,以提取液对铜绿微囊藻的生长阻碍率为评价指标,通过单因素和正交试验获得提取狐尾藻粗提液的最优工艺条件:酶解时间为4h,最适p H为3.0,温度为30℃,酶用量(纤维素酶质量/藻粉质量)为21%,超声波功率为120W,超声时间为1h。无水乙醇试剂空白对铜绿微囊藻的生长无明显影响。在最优工艺条件下,超声波-纤维素酶狐尾藻提取液对铜绿微囊藻的生长阻碍率远远高于水浸提液对铜绿微囊藻的生长阻碍率,两者存在显著性差异(P0.05)。与水浸提液相比,仅低浓度组(5g/L、10g/L)在7d后对铜绿微囊藻抑制作用消失,其他各组仍保留着比水浸提液更高的抑制能力,尤其是在提取液浓度为40g/L,反应时间为24h时,超声波-纤维素酶提取液对铜绿微囊藻的最大生长阻碍率为271.74%,而水浸提液对铜绿微囊藻的生长阻碍率只有129.78%。超声波-纤维素酶狐尾藻提取液作用于铜绿微囊藻,随着反应时间的延长和提取液浓度的升高,铜绿微囊藻T-SOD活力、CAT活力和MDA含量12h内快速下降,12h后下降速率减慢,尤其是高浓度组(30g/L、40g/L)T-SOD活力、CAT活力和MDA含量96h时趋于0;随着浓度升高,T-SOD活力、CAT活力和MDA含量与提取液浓度呈负相关,与对照组相比,各浓度组均存在显著性差异(P0.05)。狐尾藻水浸提液和超声波-纤维素酶提取液对斑马鱼急性毒性研究中,水浸提液和超声波-纤维素酶提取液各浓度组斑马鱼存活率均高于对照组存活率,尤其是超声波-纤维素酶提取液各浓度组斑马鱼存活率在反应120h内均为100%,对斑马鱼存活率无影响。根据化学物质对鱼类和水蚤的毒性等级评价可知,水浸提液和超声波-纤维素酶提取液对斑马鱼均为低毒性物质。因此,超声波-纤维素酶提取液对于生态系统是安全的,可被用于生产实践中去除水华,保护水生生态系统,保护环境。
[Abstract]:In recent years, environmental pollution is becoming more and more serious, eutrophication of water is becoming more and more serious, and Shui Hua is frequent. Microcystis aeruginosa, as a common Shui Hua cyanobacteria, has brought serious threat to people's life and the development of industry and agriculture. In this study, the allelopathic substances in the stem of Myriophyllum spicatum were studied, and the optimum conditions for extracting allelochemicals from Myriophyllum spicatum by ultrasonic and cellulase method were analyzed. To study the effect of extract on antioxidant enzyme system of microcystis aeruginosa cells and acute toxicity of zebrafish. It was found that the ultrasonic cellulase extract could inhibit the growth of microcystis aeruginosa quickly and effectively, destroy the antioxidant enzyme system of algal cells and kill the algal cells, but it was a low toxic substance to zebrafish. It provides some theoretical support for further research in the future. Ultrasonic wave cellulase method was used to extract the anti-algal components of Microcystis aeruginosa. The inhibitory rate of the extract on the growth of Microcystis aeruginosa was used as the evaluation index. Through single factor and orthogonal experiment, the optimum conditions of extracting crude extract of Sargassum foxellus were obtained: the enzymatic hydrolysis time was 4 h, the optimum pH was 3.0, the temperature was 30 鈩，

本文编号：1900100