Application of Acetyl-CoA synthetase from Methanothermobacter thermautotrophicus to non-native substrates
Application of Acetyl-CoA synthetase from Methanothermobacter thermautotrophicus to non-native substrates
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DOI：
10.1016/j.enzmictec.2019.05.005
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发表时间：
2019-09-01
影响因子：
3.4
通讯作者：
Stewart, Jon D.
中科院分区：
文献类型：
--
作者：
Mouterde, Louis M. M.;Stewart, Jon D.
研究了嗜热甲烷杆菌乙酰辅酶A合成酶Trp（416）Gly突变体（Trp（416）Gly MT-ACS 1）的底物选择性。目标是确定其底物范围，特别是官能化羧酸底物，其允许使用复分解或点击化学对CoA硫酯或下游产物进行合成后官能化。通过原位形成酰基异羟肟酸铁（III）络合物测定相对活性。Trp（416）Gly MT-ACS 1对C-2至C-8的饱和直链羧酸、C-4至C-7的ω-烯基直链羧酸和C-5至C-7的ω-炔基直链羧酸显示出良好的活性。在筛选反应中显示>= 20%转化率的羧酸用于完全消耗所添加CoASH的制备转化。
The substrate selectivity of the Trp(416)Gly mutant of Methanothermobacter thermautotrophicus acetyl-CoA synthetase (Trp(416)Gly MT-ACS1) was explored. The goal was to identify its substrate range, particularly for functionalized carboxylic acid substrates that would allow post-synthesis functionalization of CoA thioesters or downstream products using metathesis or Click chemistry. Relative activities were determined by in situ formation of acyl-hydroxamate iron (III) complexes. Trp(416)Gly MT-ACS1 showed good activities for saturated straight chain carboxylic acids from C-2 to C-8, for omega-alkenyl straight chain carboxylic acids from C-4 to C-7 and for co-alkynyl straight chain carboxylic acids from C-5 to C-7. Carboxylic acids showing >= 20% conversion in screening reactions were used in preparative conversions that completely consumed the added CoASH.