摘要:本设计分为两部分，其一为茄子中提取的过氧化物酶的分离纯化，其二为茄子过氧化物酶的理化性质的研究。为优化茄子过氧化物酶的提取及分离纯化工艺，本设计通过匀浆破碎法、硫酸盐析、透析、离子交换层析、分子筛柱层析对茄子过氧化物酶逐步提纯，排除了其他杂质对过氧化物酶纯度的影响，获得了提取茄子中过氧化物酶的成熟工艺，以吸光度作为衡量茄子过氧化物酶浓度高低的指标，结果表明：经上述各步分离纯化后最终酶茄子过氧化物酶分子质量大小约为53.3KDa，比活力为324.444U/mg，纯化倍数为294.950, 可以看出经过多步分离纯化后，得到了纯度较高的茄子过氧化物酶制品。为了了解茄子中提取的过氧化物酶的理化性质，本设计在获得成熟提取分离纯化茄子过氧化物酶的基础上考察了茄子过氧化物酶的最适反应温度、热稳定性、最适反应pH、酸碱稳定性、金属离子对其的影响。结果表明：茄子过氧化物酶的最适反应温度为57℃，在温度在20℃-60℃酶活性较稳定性，最适反应pH为6，对于酸碱稳定性较差较敏感，Zn2+，Fe2+，Cu2+对茄子过氧化物酶有较好的激活作用；Ca2+，Mg2+，Mn2+，Al3+，Ba2+，Na+，K+等对该酶有较强的抑制作用，表型活性剂对茄子的过氧化物酶的影响很小。
关键词 茄子；过氧化物酶；分离纯化；理化性质
Abstract:The design was divided into two parts, the one was the separation and purification of peroxidase from eggplant, and the second was for the study of the physical and chemical properties of eggplant peroxidase. In order to optimize the separation and purification of the eggplant peroxidase. The peroxidase was purified through sulfate precipitation, dialysis, ion exchange chromatography, molecular sieve chromatography, and it gradually excluded other impurities on the purity of peroxidase, the results showed that: after the step-by-step separation and purification, the molecular mass of the enzyme was 53.3KDa, the final enzyme activity was 324.444U/mg, and the purification factor was 294.950. It could be seen high purity eggplant peroxidase products after a multistep separation and purification. In order to understand the physical and chemical properties of peroxidase which was extracted from the eggplant, the design investigated the effects of the eggplant peroxidase optimum reaction temperature, thermal stability, optimum reaction pH, pH stability, metal ions a surface-active agent. The results showed that: The optimum reaction temperature of eggplant peroxidase was 57℃. The enzyme was stable under 20~60℃. The optimum reaction pH was 6. Its activity could be strongly activated byZn2+, Fe 2+and Cu2+ , but inhibited by Ca2+, Mg2+, Mn2+, Al3+, Ba2+, Na+, K+.
Keywords Eggplant Peroxidase Separation and purification Physical and chemical properties
