PLA降解菌论文巨大芽孢杆菌(Bacillus megaterium)的选育及其PLA降解酶的研究.docVIP

PLA降解菌论文：巨大芽孢杆菌（Bacillus megaterium）的选育及其PLA降解酶的研究 【中文摘要】随着对塑料制品需求的不断增加,带来了不可忽视的能源危机和环境危机。普通塑料在自然环境下几十年都不能被降解,因此开发可降解塑料迫在眉睫。目前开发的可降解塑料主要是聚酯类物质,而其中PLA以其优良的物理化学性能和潜在的成本优势尤受人们的关注。虽然PLA在生产和应用上有比较明显的优势,但是它的生物降解研究比较滞后。因此,本文从PLA降解菌株出发,从菌株的筛选到鉴定,然后测定其最适产酶条件,并进行PLA降解酶的分离纯化。本文主要的工作内容及结果如下:1.对辽宁省抚顺市抚顺石油一厂污水处理车间曝气池中的活性污泥进行PLA降解菌株的筛选。获得了一株能够降解PLA的菌株编号为DS04—W。2.对筛选的DS04—W进行形态学观察,生理生化性质测定,以及16S rDNA序列的测定,最后鉴定该菌为Bacillus megaterium(巨大芽孢杆菌)。3.对DS04—W菌降解PLA膜能力的测定,用扫描电镜观察PLA膜降解前后的形态变化。发现在降解过程中透明度逐渐下降,表面变得粗糙,并且肉眼可见细微小泡突起。该结果证明DS04-W菌株确实对PLA膜具有降解能力。4.最适产酶条件的测定,首先从明胶、酪氨酸、酪蛋白、酵母粉、蛋白胨中选出最佳的诱导物,根据实验结果选定为酵母粉;然后通过对温度、pH值、摇床转数、接种量单因素水平进行测定,再进行四因素三水平的正交实验,确定最佳发酵条件为培养时间为48h,培养基起始pH为8.5,摇床转速为200r/min,种子培养基接种量为6%(V/V)。 【英文摘要】Along with the increased to plastic products demand, it has brought the noticeabl e energy crisis and the environment crisis. The ordinary plastic cannot be degraded in the natural environment for decades, therefore it is important to develop biodegradabl e plastic.The development of biodegradable plastic mainly include polyesters, among all polyesters, poly(L-Lactic acid) (PLA) is especially promising with its excellent physicochemical properties and the potential price benefit. Although PLA has distinct advantages in production and application over other polyesters, the study of its biodegradation is relatively lagging behind its synthesis. In this thesis, we screening and appraisal from strains, determine the optimal conditions of producting enzyme , isolate and purify of PLA梔egrading enzymes. The major results of the thesis are as follows:1. Poly (L-lactic acid) (PLA)-degrading strain, was isolated by screening activated sludge produced from Fu shun petroleum factory. The PLA strain is numbered for DS04 - W.2. The DS04梂 is observed on morphological, physiological and biochemical properties mensuration, and determine 16S rDNA sequence. Finally the DS04梂 is identified the bacterium Bacillus megaterium.3.The DS04梂 bacteria is determined the degradation ability of PLA film.