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壬基酚降解酶的分离纯化及性质研究
  • ISSN号:1009-6094
  • 期刊名称:《安全与环境学报》
  • 时间:0
  • 分类:X17[环境科学与工程—环境科学]
  • 作者机构:[1]东华大学环境科学与工程学院,上海201620
  • 相关基金:国家自然科学基金项目(20677008);上海市重点学科建设项目(B604)
中文摘要:

从恶臭假单胞菌(Pseudomonas putida)X-8中分离纯化出一种壬基酚降解酶,研究其酶学特性。将该菌株X-8在NP-尿素培养基中振荡培养26h(30℃),其细胞经超声波破壁,得到粗提酶液。粗提酶液经硫酸铵分级盐析、透析脱盐、Sephadex G-200分子筛柱层析、DEAE—Sepharose Fast Flow离子交换层析,得到凝胶电泳均一的NP降解酶。SDS—PAGE电泳结果表明,该酶相对分子质量为37000±1,适宜作用条件为30℃、pH=7.0。该酶在pH=7.0—10.0、10—50℃时非常稳定。本研究得到的壬基酚降解酶活力强,常温下反应速度快、稳定性高,可用于污水中壬基酚的生物降解。将壬基酚降解酶纯化至电泳纯时工艺复杂、收率较低,但实际应用中只需对该酶进行粗分离,其工艺简单、收率高,因此该酶可用于含NP废水的处理。

英文摘要:

The present paper aims to introduce an approach to the purification and analysis of the characteristic features of a nonylphenol degrading enzyme from Pseudomonas putida. The nonylphenol degrading enzyme mentioned here can be purified into a SDS- PAGE homogenous state. The actual process of purification can be stated as follows: at the beginning, it was proceeding very complicated with low yield, but when used to degrade nonylphenol, the enzyme can just be sorted out in the first two steps. If so, the purification process could be made simplified with high yield. From what is said above, it carl be seen that the isolation or purification method is feasible and economically practical suppose the enzyme obtained can be applied for sewage treatment with NP. Mention is also needed that this research has been conducted based on selecting high efficient NP degradation bacteria, known as Pseudomonas put/da X - 8 from the activated sludge. The Pseudomonas put/da X - 8, cultured in urea medium at 30℃ for 26 hours, and then the with the cells collected by centrifugation at a speed of 8 000 r/min, the cell wall tends to be cracked by the ultrasonic disintegration at 4 ℃, at which the crude enzyme can be extracted with buffer. Then the crude enzyme can be purified into homogenous SDS - PAGE by ammonium sulfate precipitation, dialysis, Sephadex G - 200 gel filtration and DEAE - Sepharose Fast Flow. The result of SDS - PAGE shows that the molecular weight of the enzyme is 37 000 ± 1, and the optimal reaction factors of the enzyme proves at pH = 7.0 and 30 ℃. In such a situation, the enzyme purified remains stable at the temperature between 10- 50 ℃ with pH=7.0- 10.0.

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期刊信息
  • 《安全与环境学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国兵器工业集团公司
  • 主办单位:北京理工大学 中国环境科学学会 中国职业安全健康协会
  • 主编:冯长根
  • 地址:北京市海淀区中关村南大街5号
  • 邮编:100081
  • 邮箱:aqyhjxb@263.net;aqyhjxb@wuma.com.cn
  • 电话:010-68913997
  • 国际标准刊号:ISSN:1009-6094
  • 国内统一刊号:ISSN:11-4537/X
  • 邮发代号:2-770
  • 获奖情况:
  • 获首届《CAJ-CD》执行优秀期刊奖,中国科技论文统计源期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:17182