Purification and characterization of the aromatic desulfinase, 2-(2'-hydroxyphenyl)benzenesulfinate desulfinase.

Purification and characterization of the aromatic desulfinase, 2-(2'-hydroxyphenyl)benzenesulfinate desulfinase.
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芳香族脱硫酶、2-(2-羟苯基)苯亚磺酸脱硫酶的纯化和表征。

DOI:
10.1016/s0003-9861(03)00230-3
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发表时间:
2003
影响因子:
3.9
通讯作者:
G. T. Mrachko
G. T. Mrachko
中科院分区:
生物学3区
文献类型:
--
作者:
L. Watkins;R. Rodriguez;D. Schneider;R. Broderick;M. Cruz;R. Chambers;E. Ruckman;M. Cody;G. T. Mrachko

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2-(2′-羟基苯基)苯亚磺酸脱硫酶 (HPBS 脱硫酶) 催化 2-(2′-羟基苯基)苯亚磺酸 (HPBS) 的碳硫键断裂,形成羟基联苯和亚硫酸盐。这是二苯并噻吩脱硫的最后一步,二苯并噻吩是用于研究石油中间馏分油生物脱硫的有机硫化合物。 HPBS 脱硫酶从红球菌 IGTS8 中纯化 1600 倍。使用荧光分光光度测定和 SDS-PAGE 监测纯化情况。 HPBS脱硫酶的pI为5.6,最适温度为35℃,最适pH为7.0。 HPBS 脱硫酶的 Km 为 0.90±0.15μM,kcat 为 1.3±0.07min−1。测试了几种类似物作为 HPBS 脱硫酶底物或抑制剂的能力。没有发现替代底物和很少的抑制剂。当 Cu2+ 和 Zn2+ 存在时,HPBS 脱硫酶活性降低,而没有金属显着增强酶活性。 HPBS 脱硫酶对酪氨酸、色氨酸和半胱氨酸特异性修饰剂敏感。
2-(2′-Hydroxyphenyl)benzenesulfinate desulfinase (HPBS desulfinase) catalyzes the cleavage of the carbon–sulfur bond of 2-(2′-hydroxyphenyl)benzenesulfinate (HPBS) to form hydroxybiphenyl and sulfite. This is the final step in the desulfurization of dibenzothiophene, the organosulfur compound used to study biodesulfurization of petroleum middle distillate. HPBS desulfinase was purified 1600-fold from Rhodococcus IGTS8. The purification was monitored using a spectrofluorimetric assay and SDS–PAGE. The pI of HPBS desulfinase is 5.6, the temperature optimum is 35°C, and the pH optimum is 7.0. HPBS desulfinase has a Kmof 0.90±0.15μM and a kcatof 1.3±0.07min−1. Several analogs were tested for their ability to act as substrates or inhibitors of HPBS desulfinase. No alternative substrates and very few inhibitors were identified. HPBS desulfinase activity decreases in the presence of Cu2+and Zn2+, while no metals significantly enhance enzyme activity. HPBS desulfinase is susceptible to tyrosine, tryptophan, and cysteine specific modification agents.
半胱氨酸 42 对于维持 O-乙酰丝氨酸硫化氢解酶的完整活性位点非常重要,从而导致 α-氨基丙烯酸酯中间体的稳定。
DOI: 10.1021/bi980647k
发表时间: 1998
期刊: Biochemistry.
影响因子: --
作者:
Tai,CH;Yoon,MY;Kim,SK;Rege,VD;Nalabolu,SR;Kredich,NM;Schnackerz,KD;Cook,PF
通讯作者: Cook,PF
DOI: 10.1016/s0163-7258(97)00099-5
发表时间: 1997-10
影响因子: 13.5
作者:
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通讯作者: W. Montfort;A. Weichsel
DOI: 10.1021/bi00020a002
发表时间: 1995-05-23
期刊: BIOCHEMISTRY
影响因子: 2.9
作者:
FISHER, AJ;RAUSHEL, FM;RAYMENT, I
通讯作者: RAYMENT, I
Cystalysin,一种来自密螺旋体的 46 kDa L-半胱氨酸脱硫酶:生化和生物物理特征。
DOI: 10.1086/515164
发表时间: 1999
期刊: Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.
影响因子: --
作者:
Chu,L;Ebersole,JL;Kurzban,GP;Holt,SC
通讯作者: Holt,SC