期刊论文

Expression, purification, and characterization of the recombinant hydroxyquinol-1, 2-dioxygenase from Pseudomonas sp.strain.

中文

华中师范大学学报(自然科学版)

1000-1190

2015

49

5

758-762

10.19603/j.cnki.1000-1190.2015.05.018

31371893:国家自然科学基金 31071653:国家自然科学基金 31101595:国家自然科学基金

本校为第一机构

由pnpC编码的偏苯三酚1,2-双加氧酶(Hydroxyquinol 1,2-dioxygenase,PnpC)是微生物分解硝基芳香族类环境污染物的关键酶.本研究从对硝基苯酚(p-nitrophenol,PNP)降解菌HS-D38(Pseudomohas sp.)中克隆pnpC基因,利用E.coli BL21 (DE3)高效表达重组PnpC,通过亲和色谱纯化并分析其催化特性.实验结果表明:HS-D36的pnpC开放阅读框长度为873 bp,编码290个氨基酸,酶蛋白相对分子量33 KD;20℃、异丙基硫代-β-半乳糖苷(Isopropyl β-D-1 thiogalactopy-ranoside,IPTG)诱导可高效表达重组PnpC;重组酶经Ni-NTA亲和色谱一步分离可达到电泳纯;纯酶比活力9.3 U/mg,纯化倍数37.2,活力收率23.8％;重组PnpC催化邻...

Hydroxyquinol 1, 2-dioxygenase （PnpC）, encoded by pnpC gene, is a key enzyme for bacteria to degrade a series of recalcitrant nitrophenol pollutants, such as p-nitrophenol （PNP）. In the present work, a pnpC gene from Pseudomonas sp. strain HS- D36 was cloned which was able to degrade PNP efficiently. The open reading frame of the pnpC contains 873 nucleotides and codes for 290 amino acids （PnpC） with the molecular weight of 33 KD. The recombinant PnpC was highly expressed in E. coli BL21 （DE3） as a histidine-tag fusion protein and purif...