工程酵母助力真菌嵌合萜类合酶的快速系统挖掘

doi:10.12211/2096-8280.2021-084

合成生物学 ›› 2021, Vol. 2 ›› Issue (5): 666-673.DOI: 10.12211/2096-8280.2021-084

工程酵母助力真菌嵌合萜类合酶的快速系统挖掘

范震¹^,², 潘海学¹^,², 唐功利¹^,²

^1.中国科学院大学杭州高等研究院，化学与材料科学学院，浙江杭州 310024
^2.中国科学院上海有机化学研究所，生命有机化学国家重点实验室，上海 200032

收稿日期:2021-08-13 修回日期:2021-08-29 出版日期:2021-11-19 发布日期:2021-11-19
通讯作者: 唐功利
作者简介:范震（1987—），男，博士，博士后，主要从事天然产物生物合成研究。E-mail：fanzhen@sioc.ac.cn
唐功利（1971—），男，博士，研究员，主要从事天然产物生物合成和化学生物学研究。E-mail：gltang@sioc.ac.cn
基金资助:
国家自然科学基金重点项目(31930002)

Engineered yeast facilitates rapid and systematic mining of fungal chimeric terpene synthases

Zhen FAN¹^,², Haixue PAN¹^,², Gongli TANG¹^,²

^1.School of Chemistry and Materials Science，Hangzhou Institute for Advanced Study，University of Chinese Academy of Sciences，Hangzhou 310024，Zhejiang，China
^2.State Key Laboratory of Bio-Organic and Natural Products Chemistry，Shanghai Institute of Organic Chemistry，Chinese Academy of Sciences，Shanghai 200032，China

Received:2021-08-13 Revised:2021-08-29 Online:2021-11-19 Published:2021-11-19
Contact: Gongli TANG

摘要/Abstract

摘要：

真菌特有的嵌合萜类合酶（简称为PTTS）由C端异戊烯基转移酶（PT）结构域和N端I型萜类合酶（TS）结构域组成，可直接催化异戊二烯单元产生结构多样的二萜和二倍半萜化合物。自2007年发现第一个PTTS以来，只有约20个PTTS的功能被验证，人们对PTTS的起源和功能进化的认识还十分有限。最近武汉大学刘天罡教授课题组与美国田纳西大学陈峰教授课题组合作，运用高效萜类前体供给的酿酒酵母底盘并结合高通量的自动化平台，对PTTS的起源和功能进化进行了深入系统的探索，扩充了嵌合萜类合酶家族成员的数量，并为萜类化合物的挖掘提供了一个高效的方法。

关键词: 生物合成, 嵌合萜类合酶, 进化, 萜类, 酵母底盘

Abstract:

Chimeric terpene synthases (PTTSs), which consist of C-terminal prenyltransferase (PT) and N-terminal ClassⅠterpene synthase (TS) domains, are unique to fungi for catalyzing the synthesis of structurally diverse diterpenes and sesterterpenes. Since the first PTTS was discovered in 2007, only about 20 PTTSs have been functionally verified, and understanding of the origin and functional evolution of PTTS genes is very limited. Recently, Professor Tiangang Liu's research team at Wuhan University and Professor Feng Chen's research team at the University of Tennessee in the United States have conducted in-depth exploration on the origin and functional evolution of PTTS with an efficient terpene precursor yeast chassis system combined with the high-throughput automatic screening platform. The research not only expands the chimeric terpene synthase family, but also provides an advanced method for mining more terpene synthases for the biosynthesis of terpenoids.

Key words: biosynthesis, chimeric terpene synthase, evolution, terpenoids, yeast chassis

中图分类号:

范震, 潘海学, 唐功利. 工程酵母助力真菌嵌合萜类合酶的快速系统挖掘[J]. 合成生物学, 2021, 2(5): 666-673.

Zhen FAN, Haixue PAN, Gongli TANG. Engineered yeast facilitates rapid and systematic mining of fungal chimeric terpene synthases[J]. Synthetic Biology Journal, 2021, 2(5): 666-673.

图/表 6

图1 异戊烯基转移酶（PT）和萜类合酶（TS）催化萜类基本骨架形成的简略过程

Fig. 1 Formation of terpenoid skeletons through the catalysis by isopentenyl transferase (PT) and terpene synthase (TS)

图2 Ⅰ型和Ⅱ型萜类合酶（TS）催化环化的过程的两例

Fig. 2 Two cases for cyclization catalyzed by TypeⅠand TypeⅡterpene synthases (TSs)

图3 4种嵌合萜类合酶催化反应范例

Fig. 3 Catalytic reactions of 4 chimeric terpene synthases

图4 PTTS催化的两种环化类型

Fig. 4 Two cyclization reactions catalyzed by PTTS

图5 代谢工程结合化学合成的方法合成青蒿素和(-)-englerin A

Fig. 5 Synthesis of artemisinin and (-)-englerin A by metabolic engineering combined with synthetic chemistry

图6 54个PTTS催化合成的萜类化合物及其结构(红色标记为之前报道的PTTS及其催化合成的化合物结构；粉红色标记为刘天罡课题组新鉴定的24个PTTS及其催化合成的化合物结构，其中黑体加粗标记为新化合物，绿色标记为已报道的植物来源化合物，蓝色标记为已报道的微生物来源化合物)

Fig. 6 Terpenoids and their structures synthesized under the catalysis of 54 PTTSs(Previously reported PTTSs and resulting products are marked in red, and 24 newly identified PTTSs by Liu Tiangang et al and resulting products are marked in pink. Products from plants and microorganisms are marked in green and blue, respectively)

参考文献 23

1	Dictionary of natural products[EB/OL]. .
2	CHRISTIANSON D W. Structural and chemical biology of terpenoid cyclases[J]. Chemical Reviews, 2017, 117(17): 11570-11648.
3	GERSHENZON J, DUDAREVA N. The function of terpene natural products in the natural world[J]. Nature Chemical Biology, 2007, 3(7): 408-414.
4	CHEN M B, CHOU W K W, TOYOMASU T, et al. Structure and function of fusicoccadiene synthase, a hexameric bifunctional diterpene synthase[J]. ACS Chemical Biology, 2016, 11(4): 889-899.
5	GAO J, KO T P, CHEN L, et al. "Head-to-Middle" and "Head-to-Tail" cis-Prenyl transferases: structure of isosesquilavandulyl diphosphate synthase[J]. Angewandte Chemie-International Edition, 2018, 57(3): 683-687.
6	SALLAUD C, RONTEIN D, ONILLON S, et al. A novel pathway for sesquiterpene biosynthesis from Z,Z-farnesyl pyrophosphate in the wild tomato Solanum habrochaites[J]. The Plant Cell, 2009, 21(1): 301-317,
7	CHRISTIANSON D W. Roots of biosynthetic diversity[J]. Science, 2007, 316(5812): 60-61.
8	THOLL D. Terpene synthases and the regulation, diversity and biological roles of terpene metabolism[J]. Current Opinion in Plant Biology, 2006, 9(3): 297-304.
9	THOLL D. Biosynthesis and biological functions of terpenoids in plants[J]. Biotechnology of Isoprenoids, 2015, 148: 63-106
10	MITSUHASHI T, ABE I. Chimeric terpene synthases possessing both terpene cyclization and prenyltransfer activities[J]. ChemBioChem, 2018, 19(11): 1106-1114.
11	KANG W, MA T, LIU M, et al. Modular enzyme assembly for enhanced cascade biocatalysis and metabolic flux[J]. Nature Communications, 2019, 10(1): 4248.
12	FAYLO J L, EEUWEN T VAN, KIM H J, et al. Structural insight on assembly-line catalysis in terpene biosynthesis[J]. Nature Communications, 2021, 12(1): 3487.
13	MINAMI A, OZAKI T, LIU C W, et al. Cyclopentane-forming di/sesterterpene synthases: widely distributed enzymes in bacteria, fungi, and plants[J]. Natural Product Reports, 2018, 35(12): 1330-1346.
14	TOYOMASU T, TSUKAHARA M, KANEKO A, et al. Fusicoccins are biosynthesized by an unusual chimera diterpene synthase in fungi[J]. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(9): 3084-3088.
15	CHIBA R, MINAMI A, GOMI K, et al. Identification of ophiobolin F synthase by a genome mining approach: a sesterterpene synthase from Aspergillus clavatus[J]. Organic Letters, 2013, 15(3): 594-597.
16	CHEN R, JIA Q D, MU X, et al. Systematic mining of fungal chimeric terpene synthases using an efficient precursor-providing yeast chassis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2021, 118(29): e2023247118.
17	YE Y, MINAMI A, MANDI A, et al. Genome mining for sesterterpenes using bifunctional terpene synthases reveals a unified intermediate of di/sesterterpenes[J]. Journal of the American Chemical Society, 2015, 137(36): 11846-11853.
18	MATSUDA Y, MITSUHASHI T, LEE S, et al. Astellifadiene: structure determination by NMR spectroscopy and crystalline sponge method, and elucidation of its biosynthesis[J]. Angewandte Chemie-International Edition, 2016, 55(19): 5785-5788.
19	ZHU F Y, ZHONG X F, HU M Z, et al. In vitro reconstitution of mevalonate pathway and targeted engineering of farnesene overproduction in Escherichia coli[J]. Biotechnology and Bioengineering, 2014, 111(7): 1396-1405.
20	RO D K, PARADISE E M, OUELLET M. et al. Production of the antimalarial drug precursor artemisinic acid in engineered yeast[J]. Nature, 2006, 440(7086): 940-943.
21	KAMPRANIS S C, MAKRIS A M. Developing a yeast cell factory for the production of terpenoids[J]. Computational and Structural Biotechnology Journal, 2012, 3: e201210006.
22	PADDON C J, WESTFALL P J, PITERA D J, et al. High-level semi-synthetic production of the potent antimalarial artemisinin[J]. Nature, 2013, 496(7446): 528-532.
23	SIEMON T, WANG Z Q, BIAN G K, et al. Semisynthesis of plant-derived englerin A enabled by microbe engineering of guaia-6,10(14)-diene as building block[J]. Journal of the American Chemical Society, 2020, 142(6): 2760-2765.

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工程酵母助力真菌嵌合萜类合酶的快速系统挖掘

Engineered yeast facilitates rapid and systematic mining of fungal chimeric terpene synthases

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