多酶催化串联策略在复杂天然产物合成中的应用

doi:10.12211/2096-8280.2020-010

摘要/Abstract

摘要：

结构新颖、复杂多样的天然产物及其衍生物一直是新药发现的重要源泉。然而，传统的从动植物及微生物中提取分离天然产物的方法往往存在效率低、耗时长、成本高等问题，极大地制约了新型药物的研发。天然产物的全合成主要包括化学合成和生物合成，其中体外酶催化合成因其具有催化效率高、反应选择性强、目标产物专一、反应条件简单温和、绿色环保等优点，为复杂天然产物的合成提供了有效策略。本文总结了近年来采用体外酶催化方法合成复杂天然产物的研究进展，着重介绍了多酶催化串联方法在聚酮类、生物碱类、萜类、甾体类、大环内酰胺类、核苷类等复杂天然产物合成中的应用，并对其目前存在的问题以及解决对策进行了讨论，同时对其未来发展也进行了展望。

关键词: 天然产物, 生物合成, 酶催化, 串联

Abstract:

Structurally diverse natural products and their derivatives have been an indispensable source for the discovery of new drugs. However, the traditional extraction, isolation and characterization of natural products from animals, plants and microorganisms often have challenges such as low efficiency, time-consuming and expensive cost, which greatly hinder their development as new drugs. The complete synthesis of natural products mainly includes chemical synthesis and biosynthesis. Among them, In vitro enzymatic synthesis has the advantages of high catalytic efficiency, high chemo-, stereo- and regio-selectivity, specific product spectrum, simple and mild reaction conditions and environmental friendly, which make it an effective strategy for the synthesis of complex natural products. This paper reviews recent research advances in the synthesis of complex natural products by in vitro enzymatic methods, especially the applications of the multienzyme-catalyzed tandem strategy for synthesis of complex natural products including polyketides, alkaloids, terpenoids, steroids, macrolides and nucleosides. Furthermore, the bottlenecks, corresponding solutions and the perspective of this strategy are discussed.

Key words: natural products, biosynthesis, enzymatic catalysis, multienzyme-catalyzed tandem strategy

中图分类号:

Q814.9

贺俊斌, 孟松, 潘海学, 唐功利. 多酶催化串联策略在复杂天然产物合成中的应用[J]. 合成生物学, 2020, 1(2): 226-246.

Junbin HE, Song MENG, Haixue PAN, Gongli TANG. Applications of the multienzyme-catalyzed tandem strategy in the synthesis of complex natural products[J]. Synthetic Biology Journal, 2020, 1(2): 226-246.

图/表 14

图1 多酶催化串联法合成肠道菌素（1）和wailupemycins[SAM，S-腺苷蛋氨酸（S-adenosyl methionine）；NADPH，烟酰胺腺嘌呤二核苷酸磷酸（nicotinamide adenine dinucleotide phosphate）；ATP，三磷酸腺苷（adenosine triphosphate）；ferredoxin，铁氧还蛋白；ferredoxin-NADP+ reductase，铁氧还蛋白- NADP+还原酶；catalase，过氧化氢酶]

Fig. 1 Multienzyme-catalyzed tandem synthesis of enterocin (1) and wailupemycins

图2 Gilvocarcin V和司替霉素的结构式及多酶催化串联法合成腊伯罗霉素（5）、defucogilvocarcin M（6）和presteffimycinone（7）[flavin reductase，黄素还原酶；FAD，黄素腺嘌呤二核苷酸(flavin adenine dinucleotide)]

Fig. 2 Chemical structures of gilvocarcin V and steffimycin and the multienzyme-catalyzed tandem synthesis of rabelomycin (5), defucogilvocarcin M (6) and presteffimycinone (7)

图3 多酶催化串联法合成赛洛西宾（8）

Fig. 3 Multienzyme-catalyzed tandem synthesis of psilocybin (8)

图4 多酶催化串联法合成苄基异喹啉生物碱[（9）～（16）]

Fig. 4 Multienzyme-catalyzed tandem synthesis of benzylisoquinoline alkaloids [(9)～(16)]

图5 Thaxtomins的生物合成途径（L-arginine，L-精氢酶）

Fig. 5 Biosynthetic pathway of thaxtomins

图6 双环霉素（22）的生物合成途径（a）和体外酶催化合成（b）（α-ketoglutarate，α-酮戊二酸；L-ascorbic acid，抗坏血酸）

Fig. 6 Biosynthetic pathway (a) and in vitro multienzyme-catalyzed synthesis (b) of bicyclomycin (22)

图7 多酶催化串联法合成赤霉素GA4（23）[ACS,乙酰辅酶A合成酶（acetyl-CoA synthase）；AACT,乙酰乙酰辅酶A硫解酶（acetoacetyl-CoA thiolase）；HMGS,羟甲基戊二酰辅酶A合成酶（hydroxymethylglutaryl-CoA synthase）；HMGR,羟甲基戊二酰辅酶A还原酶（hydroxymethylglutaryl-CoA reductase）；MVK,甲羟戊酸激酶（mevalonate kinase）；PMVK,磷酸甲羟戊酸激酶（phosphomevalonate kinase）；DMDC,二磷酸甲羟戊酸脱羧酶（diphosphomevalonate decarboxylase）；IPI,异戊烯基焦磷酸异构酶（isopentenyl diphosphate isomerase）；GGPS,牻牛儿基牻牛儿基焦磷酸合成酶（geranylgeranyl diphosphate synthase）]

Fig. 7 Multienzyme-catalyzed tandem synthesis of gibberellin A4 (23)

图8 多酶催化串联法合成熊去氧胆酸（24）（lactate，乳酸盐；pyruvate，丙酮酸盐；gluconate，葡萄糖酸盐；glucose，葡萄糖）

Fig. 8 Multienzyme-catalyzed tandem synthesis of ursodeoxycholic acid (24)

图9 斑鸠霉素（25）的生物合成途径

Fig. 9 Biosynthetic pathway of ikarugamycin (25)

图10 多酶催化串联法合成UK-2A类似物

Fig. 10 Multienzyme-catalyzed tandem synthesis of UK-2A analogues

图11 多酶催化串联法合成islatravir（41）（fructose，果糖；glucose 1-phosphate，葡萄糖-1-磷酸）

Fig. 11 Multienzyme-catalyzed tandem synthesis of islatravir (41)

图12 酶催化串联法合成的一些其他化合物（45～57）

Fig. 12 Enzymatic tandem synthesis of other compounds 45～57

图13 化学-酶催化串联法合成多杀菌素A（58）

Fig. 13 Chemoenzymatic synthesis of spinosyn A (58)

图14 化学-酶催化串联法合成超低分子量肝素（68）和（69）

Fig. 14 Chemoenzymatic synthesis of ultralow molecular weight heparins （68） and （69）

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