4-Coumarate coenzyme A ligase and 4-hydroxybenzoate coenzyme A ligase from the hornwort Anthoceros agrestis expressed in E. coli were characterized on biochemical and molecular levels and showed interesting substrate specificities. Acyl-activating enzymes are associated with the biosynthesis or degradation of various metabolic products such as lipids, amino acids, sugars, and natural compounds. In this work, cDNA sequences encoding 4-coumarate coenzyme A ligase (4CL) and 4-hydroxybenzoate coenzyme A ligase (4HBCL) were amplified from the hornwort Anthoceros agrestis. The coding sequences were expressed in E. coli and purified by Ni-chelate chromatography. The CoA ligases exhibited different substrate specificities. 4CL catalyzed the activation of 4-coumaric acid, 3-coumaric acid, 2-coumaric acid, caffeic acid, isoferulic acid, ferulic acid, and cinnamic acid but lacked activities towards sinapic acid and benzoic acids. In contrast, 4HBCL preferred 4-hydroxybenzoic acid and benzoic acid, but also accepted other benzoic acid derivatives except salicylic acid and 3-aminosalicylic acid. Furthermore, 4HBCL also activated isoferulic acid, cinnamic acid, 2-coumaric acid, 3-coumaric acid, 4-coumaric acid and caffeic acid, but lacked affinity for ferulic acid and sinapic acid. These substrate specificities could be related to the phenolic compounds identified in Anthoceros agrestis.

中文翻译：

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金鱼藻中的酚类代谢：4-香豆酸 CoA 连接酶和 4-羟基苯甲酸 CoA 连接酶。

来自在大肠杆菌中表达的金银花 Anthoceros agrestis 的 4-香豆酸辅酶 A 连接酶和 4-羟基苯甲酸辅酶 A 连接酶在生化和分子水平上进行了表征，并显示出有趣的底物特异性。酰基活化酶与各种代谢产物如脂质、氨基酸、糖和天然化合物的生物合成或降解有关。在这项工作中，编码 4-香豆酸辅酶 A 连接酶 (4CL) 和 4-羟基苯甲酸辅酶 A 连接酶 (4HBCL) 的 cDNA 序列从金银花中扩增。编码序列在大肠杆菌中表达并通过镍螯合色谱法纯化。CoA 连接酶表现出不同的底物特异性。4CL催化4-香豆酸、3-香豆酸、2-香豆酸、咖啡酸、异阿魏酸、阿魏酸的活化，和肉桂酸，但对芥子酸和苯甲酸缺乏活性。相比之下，4HBCL首选4-羟基苯甲酸和苯甲酸，但也接受除水杨酸和3-氨基水杨酸外的其他苯甲酸衍生物。此外，4HBCL还能活化异阿魏酸、肉桂酸、2-香豆酸、3-香豆酸、4-香豆酸和咖啡酸，但对阿魏酸和芥子酸缺乏亲和力。这些底物特异性可能与 Anthoceros agrestis 中鉴定的酚类化合物有关。2-香豆酸、3-香豆酸、4-香豆酸和咖啡酸，但对阿魏酸和芥子酸缺乏亲和力。这些底物特异性可能与 Anthoceros agrestis 中鉴定的酚类化合物有关。2-香豆酸、3-香豆酸、4-香豆酸和咖啡酸，但对阿魏酸和芥子酸缺乏亲和力。这些底物特异性可能与 Anthoceros agrestis 中鉴定的酚类化合物有关。