A Thermophilic Biofunctional Multienzyme Cascade Reaction for Cell-Free Synthesis of Salvianic Acid A and 3,4-Dihydroxymandeiic Acid
A Thermophilic Biofunctional Multienzyme Cascade Reaction for Cell-Free Synthesis of Salvianic Acid A and 3,4-Dihydroxymandeiic Acid
复制标题
嗜热生物功能多酶级联反应无细胞合成丹酚酸 A 和 3,4-二羟基扁桃酸
DOI：
10.1021/acssuschemeng.9b05496
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发表时间：
2019-11-18
影响因子：
8.4
通讯作者：
Wang, Jian
中科院分区：
文献类型：
--
作者：
Li, Chao;Zhang, Zhendong;Wang, Jian
高价值酚酸的生产对制药业的可持续性具有重要意义。本研究以低成本的苯丙酮酸(PPA)和2-苯乙氧基酸(PGA)为双底物，构建了由D-扁桃酸脱氢酶(ManDH)、苯丙氨酸4-羟基酶(PAH)和羟基苯乙酸3-羟基酶(HpaH)组成的人工亲热反应级联反应体系。PPA和PGA分别以理论产率的84.9%和90.9%合成了二羟基酚酸丹参酸A(SAA)和3，4-二羟基扁桃酸(DOMA)。反应成功的关键是来自巴氏热球菌的新型TbManDH和来自热单胞菌Curvata的TcPAH表现出广泛的底物专一性。辅因子6，7-二甲基-5，6，7，8-四氢蝶呤(DMPH4)的新原位再生和烟酰胺腺嘌呤二核苷酸(NADH)的回收也有助于提高产量。考虑到许多其他天然酶广泛的底物专一性，设计这样的双功能甚至多功能催化剂系统来高效地生产增值化学品是很有吸引力的。
The production of high-value phenolic acids is of significant importance for sustainability in the pharmaceutical industry. In this study, an artificial thermophilic reaction cascade composed of D-mandelate dehydrogenase (ManDH), phenylalanine 4-hydroxylase (PAH), and hydroxyphenylacetate 3-hydroxylase (HpaH) was constructed to utilize the low-cost phenylpyruvic acid (PPA) and 2-phenylglyoxylic acid (PGA) as dual substrates. The dihydroxyphenolic acids salvianic acid A (SAA) and 3,4-dihydroxymandelic acid (DOMA) were then produced at 84.9% and 90.9% of the theoretical yields from PPA and PGA, respectively. The key to success of the reaction was the novel TbManDH from Thermococcus barophilus and TcPAH from Thermomonospora curvata that exhibited a broad substrate specificity. The newly in situ regeneration of the cofactor 6,7-dimethyl-5,6,7,8-tetrahydropterine (DMPH4) in combination with nicotinamide adenine dinucleotide (NADH) recycling also contributed to the high production yields. Considering the broad substrate specificity of many other natural enzymes, it is attractive to design such dual- or even multifunctional catalyst systems for efficiently producing value-added chemicals.