Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1
Identification and characterization of a meta-cleavage product hydrolase involved in biphenyl degradation from Arthrobacter sp. YC-RL1
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参与节杆菌属联苯降解的元裂解产物水解酶的鉴定和表征。
DOI：
10.1007/s00253-019-09956-z
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发表时间：
2019-06
影响因子：
5
通讯作者：
Yan Yanchun
中科院分区：
文献类型：
--
作者：
Jia Yang;Wang Junhuan;Ren Chao;Nahurira Ruth;Khokhar Ibatsam;Wang Jiayi;Fan Shuanghu;Yan Yanchun
多氯联苯（PCBs）是一组广泛存在于环境中的持久性有机污染物（POPs）。节杆菌属YC-RL 1是一种联苯降解菌，对芳香族化合物表现出代谢多样性。从YC-RL 1菌株中克隆了一个参与联苯生物降解的2-羟基-6-氧代-6-苯基己-2，4-二烯酸（HOPDA）水解酶（BphD）基因，并在大肠杆菌BL 21（DE 3）中异源表达。对重组BphDYC-RL 1进行了纯化和鉴定。BphDYC-RL 1在45 °C和pH 7下显示出最高活性。它在很宽的温度范围内（20-50 °C）是稳定的。该酶的Km值为0.14 mM，Kcat为11.61 s-1，Vmax为0.027 U/mg。温度依赖性催化表现出在20 °C下具有转变的双相Arrhenius图。BphDYC-RL 1可被SDS、Tween 20、Tween 80、Trition X-100、DTT、CHAPS、NBS、PMSF和DEPC灭活，但对EDTA不敏感。活性位点残基的定点突变显示，催化三联体残基（Ser 115，His 275，Asp 247）的BphDYC-RL 1是必要的，其活动。对BphDYC-RL 1的研究不仅为联苯的生物降解提供了新的潜在酶源，而且有助于加深对联苯降解催化过程和机理的认识。
Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants (POPs) widely existing in the environment. Arthrobacter sp. YC-RL1 is a biphenyl-degrading bacterium that shows metabolic versatility towards aromatic compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrolase (BphD) gene involved in the biodegradation of biphenyl was cloned from strain YC-RL1 and heterologously expressed in Escherichia coli BL21 (DE3). The recombinant BphDYC-RL1 was purified and characterized. BphDYC-RL1 showed the highest activity at 45 °C and pH 7. It was stable under a wide range of temperature (20–50 °C). The enzyme had a Km value of 0.14 mM, Kcat of 11.61 s−1, and Vmax of 0.027 U/mg. Temperature dependence catalysis exhibited a biphasic Arrhenius Plot with a transition at 20 °C. BphDYC-RL1 was inactivated by SDS, Tween 20, Tween 80, Trition X-100, DTT, CHAPS, NBS, PMSF, and DEPC, but insensitive to EDTA. Site-directed mutagenesis of the active-site residues revealed that the catalytic triad residues (Ser115, His275, and Asp247) of BphDYC-RL1 were necessary for its activity. The investigation of BphDYC-RL1 not only provides new potential enzyme resource for the biodegradation of biphenyl but also helps deepen our understanding on the catalytic process and mechanism.
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DOI：
10.3390/ijerph15091880
发表时间：
2018-08-30
影响因子：
--
作者：
Petersen MS;Halling J;Jørgensen N;Nielsen F;Grandjean P;Jensen TK;Weihe P
通讯作者：
Weihe P
影响因子：
5.6
作者：
M. Torruella;M. Flawiá;C. Eisenschlos;L. M. Vedia;C. Rubinstein;H. Torres
通讯作者：
M. Torruella;M. Flawiá;C. Eisenschlos;L. M. Vedia;C. Rubinstein;H. Torres
影响因子：
4.5
作者：
F. Yadetie;E. Oveland;A. Døskeland;F. Berven;Anders Goksøyr;O. Karlsen
通讯作者：
F. Yadetie;E. Oveland;A. Døskeland;F. Berven;Anders Goksøyr;O. Karlsen
DOI：
10.1074/jbc.273.36.22943
发表时间：
1998-09
期刊：
The Journal of Biological Chemistry
影响因子：
--
作者：
S. Seah;G. Terracina;J. Bolin;P. Riebel;V. Snieckus;L. Eltis
通讯作者：
S. Seah;G. Terracina;J. Bolin;P. Riebel;V. Snieckus;L. Eltis
影响因子：
2.8
作者：
M. Sakai;E. Masai;H. Asami;K. Sugiyama;K. Kimbara;M. Fukuda
通讯作者：
M. Sakai;E. Masai;H. Asami;K. Sugiyama;K. Kimbara;M. Fukuda