以五節芒的超氧歧化酵素和過氧化酵素做為環境污染之指標的可行性研究

Abstract

本論文從事環境污染問題的研究，目的在找尋是否有酵素隨著環境污染的梯度變化而呈規則性的改變，盼能得到如是的環境污染的指標酵素。做法上以現場堪查及植被調查為前導，接下去才進行酵素研究的採樣工作，結合野外調查與實驗室分析的工作，以使實驗室的研究結果直接反映野外調查的現況。 禮樂煉銅廠及台灣金屬礦業公司，皆以煉銅污染對當地自然環境造成重大衝擊，選擇位於禮樂廠旁的子各山的一處連續性污染梯度植被，以及台金廠工廠腹地的不連續性污染分級地區，為主要研究地點。 植被調查工作以簡單梯度穿越線來決定樣區的位置，樣區調查的方法則依丹麥學派。發現五節芒已生長約一年的植株最能反映當地的土地污染的情況，是為煉銅污染的最佳指標植物。 酵素採樣工作就以五節芒幼株為材料，分別於禮樂廠進行三個系列的採樣，台金廠亦是如此，以聚丙烯醯胺膠體電泳分析法將超氧歧化酵素與過氧化酵素電泳分離其同功酵素，進行酵素活性染色，以測驗這兩種酵素是否有某同功酵素隨著採樣地點的義意而呈現規則性的變化。 五節芒的超氧歧化酵素共有四群及主要九個的同功酵素，其中四個是Mn-SODs，五個是CuZn-SODs，其中以Mn-SODIII最能展現環境污染梯度的規則性變化；過氧化酵素則有四群及主要十個的同功酵素，但無任何一個同功酵素有如是規則性的變化。 膠體解析能力亦為重要的技術問題，發現到膠體的解析不只在於分離大馱的同功酵素，尚包括濃縮微量的同功酵素，本論文的重要成果Mn-SODIII即是在15％均質膠體集中濃縮而後才得以展現出來。

The essence of this study was focused on the observing the expression of enzymes responsing to the gradient polluted environments. At first, we had made a vegetation investigation work on the polluted area, then selected the indicator plant for marker enzymes study. Due to the particles and fumes which were produced during the processure of refining the metal, the Lii-Leh Copper Smelter and Taiwan Metal Mining Company were always the largest source of pollution for local strong envirometal stress. Therefore, we selected the hillside of the Tzu-Ko Mountain adjacent the Lii-Leh Smelter and the hinterland of Taiwan Company for the essence study area. The determination of the position of stands for vegetation investigation according to the method of simple gradient transect. The study method of vegetation stand was by the Danish School. We obtained Miscanthus floridulus as the indicator plant of the polluted area and selected its one-year-growth plants for enzymes study. After some serial of harvest the Miscanthus, the identification of the activities of superoxide dismutase (SOD) and peroxidase (PO) isozymes were undergone on the 10% acrylamide gel by activity staining. And we wished to observe the activity of the marker enzymes of the Miscanthus regularly changed with the gradient polluted stands. There were four group and nine SOD isozymes activity presenting on the gel. There were four Mn-SODs and five CuZn-SODs. Among those, the Mn-SODIII activity changed apparently according to the gradient polluted environments. Peroxdase had four group and ten isozymes. But we did not find any meaningly change in the peroxdase isozymes activity. In addition, we found that the resolvability of the gel was an important factor influence the identification of isozymes. The resolvability of gel is not only on seperating a crowd of isozymes but also on concentrating low amount isozymes. The marker enzyme Mn-SODIII showed by concentrating ability of 15% homogenous gel.