Investigation of the prevalence and catalytic activity of rubredoxin-fused alkane monooxygenases (AlkBs).
Investigation of the prevalence and catalytic activity of rubredoxin-fused alkane monooxygenases (AlkBs).
复制标题
Rubredoxin稠合烷烃单加氧酶（AlkBs）的流行率和催化活性的研究。
DOI：
10.1016/j.jinorgbio.2021.111409
复制
发表时间：
2021-06
影响因子：
3.9
通讯作者：
Austin RN
中科院分区：
文献类型：
--
作者：
Williams SC;Forsberg AP;Lee J;Vizcarra CL;Lopatkin AJ;Austin RN
对了解烷烃单加氧酶（AlkB）的环境分布的兴趣导致鉴定出超过100种不同的烷烃单加氧酶（AlkB）含有共价结合或融合的红氧还蛋白。从Dietzia cinnamea的rubredoxin融合AlkB克隆为全长蛋白质和缺失rubredoxin结构域的截短蛋白质。将点突变（V91 W）引入全长蛋白质中，目的是评估推定底物通道中的空间体积如何影响选择性。基于与烷烃和烯烃底物的活性研究，rubredoxin融合AlkB相对于其rubredoxin结构域缺失对应物氧化类似范围的烷烃底物。末端烯烃的氧化生成环氧化物和末端醛。V91 W突变体催化烯烃氧化的产物具有比在野生型蛋白存在下形成的产物更高的双链-环氧化物比。这些结果与导致影响底物定位的结构变化的该突变一致。
Interest in understanding the environmental distribution of the alkane monooxygenase (AlkB) enzyme led to the identification of over 100 distinct alkane monooxygenase (AlkB) enzymes containing a covalently bound, or fused, rubredoxin. The rubredoxin-fused AlkB from Dietzia cinnamea was cloned as a full-length protein and as a truncated protein with the rubredoxin domain deleted. A point mutation (V91W) was introduced into the full-length protein, with the goal of assessing how steric bulk in the putative substrate channel might affect selectivity. Based on activity studies with alkane and alkene substrates, the rubredoxin-fused AlkB oxidizes a similar range of alkane substrates relative to its rubredoxin domain-deletion counterpart. Oxidation of terminal alkenes generated both an epoxide and a terminal aldehyde. The products of V91W-mutant-catalyzed oxidation of alkenes had a higher aldehyde-to-epoxide ratio than the products formed in the presence of the wild type protein. These results are consistent with this mutation causing a structural change impacting substrate positioning.
登录
查看更多内容
影响因子：
4.4
作者：
Julsing, Mattijs K.;Schrewe, Manfred;Buehler, Bruno
通讯作者：
Buehler, Bruno
影响因子：
4.4
作者：
Hamamura, N;Yeager, CM;Arp, DJ
通讯作者：
Arp, DJ
影响因子：
13.6
作者：
Leticia Colin, Veronica;Fernanda Castro, Maria;Beatriz Villegas, Liliana
通讯作者：
Beatriz Villegas, Liliana
影响因子：
4.1
作者：
Gloeckner, Frank Oliver;Yilmaz, Pelin;Ludwig, Wolfgang
通讯作者：
Ludwig, Wolfgang
影响因子：
2.6
作者：
Belhaj, A;Desnoues, N;Elmerich, C
通讯作者：
Elmerich, C