摘要:海藻糖是一种非还原性双糖,是很多种生物体受环境胁迫所产生的重要的应激性代谢产物之一。本论文先对6种出发菌种进行筛选,确定安琪酿酒高活性干酵母菌株为最佳生产菌株,然后对该菌株在高渗、有毒、高温、铜等胁迫条件下海藻糖的累积进行研究,同时对海藻糖累积的最适pH值进行研究。发现在pH为6.5时最适合海藻糖的生产累积,在高渗、有毒、高温、铜胁迫单一存在时,海藻糖的积累量有大幅度上升。最适海藻糖积累的高渗条件是1%NaCl,最适有毒条件是5%乙醇,最适高温胁迫温度为42℃。但几种胁迫条件同时存在时与单一胁迫条件相比效果并不佳,证明并非环境越恶劣海藻糖累积量越多。关键词:海藻糖;酵母;胁迫7470
RESEARCH ON PRODUCTION OF TREHALOSE WITH YEAST
Abstract:The Trehalose is a non-reducing disaccharides,which is one of the important reactive metabolites produced by a lot of organisms under the stress of environment. In this paper, I first screen 6 strains, and determine that the fourth strains is the best one for producing. Then study in the hypertonic, toxic, high copper condition. At the same time, study the optimal pH of trehalose accumulation. And I get the result that in the pH of 6.5, it is most suitable for production and accumulation of trehalose; in the hypertonic, toxic, high copper condition, trehalose accumulation has jumped sharply; 1%NaCl is optimum hypertonic conditions for trehalose accumulation, the optimum toxic condition is 5% ethanol, the optimal temperature of high temperature stress is 42. Compared with the single stress condition, the effect is not good in the condition of several stress existing. So it proved that there isn’t more trehalose accumulation if the condition is worse.
key words: Trehalose; yeast; stress
目 录
1绪论 - 1 -
1.1 本课题的国内外研究进展 - 1 -
1.1.1 海藻糖的构成及性质 - 1 -
1.1.2 海藻糖的生理功能 - 1 -
1.1.3 海藻糖的国内外研究进展 - 2 -
1.2本课题的研究内容、目的及意义 - 3 -
1.2.1 海藻糖对生物质的保护作用机理 - 3 -
1.2.2海藻糖的生产方法 - 5 -
1.2.3海藻糖的应用 - 7 -
1.2.4本论文的研究内容 - 9 -
2 材料与方法 - 10 -
2.1实验材料 - 10 -
2.1.1 实验试剂 - 10 -
2.1.2 实验器材 - 12 -
2.2 分析方法: - 12 -
2.2.1 细胞干重法测定 - 12 -
2.2.2 蒽酮-硫酸法测定海藻糖含量 - 12 -
2.2.3 pH测定 - 12 -
2.2.4 酵母菌生长测定 - 12 -
2.3 实验方法 - 12 -
2.3.1 标准海藻糖曲线测定 - 12 -
2.3.2 酵母破壁方法 - 13 -
2.3.3出发菌株的筛选 - 14 -
2.3.4出发菌株的生长曲线测定 - 14 -
2.3.5 酵母内海藻糖积累的主要影响因素的研究 - 14 -
3 结果与讨论 - 15 -
3.1海藻糖标准曲线的绘制 - 15 -
3.2 酵母中海藻糖的破壁方法 - 16 -
3.3 出发菌株初筛 - 17 -
3.4 酵母菌生长曲线测定 - 18 -
3.5 酵母内海藻糖积累的主要影响因素的研究 - 19 -
3.5.1 pH值对海藻糖累积的影响 - 19 -
3.5.2 NaCl浓度对酵母菌中海藻糖累积量的影响 - 20 -
3.5.3 毒性胁迫对海藻糖累积的影响 - 21 - 发酵法生产海藻糖工艺研究+文献综述:http://www.751com.cn/shiping/lunwen_5445.html