Production of sesquiterpenoids <italic style="font-style: italic">α</italic>-neoclovene and <italic style="font-style: italic">β</italic>-caryophyllene by engineered <italic style="font-style: italic">Saccharomyces cerevisiae</italic>

LI Xiaodong; YANG Chengshuai; WANG Pingping; YAN Xing; ZHOU Zhihua

doi:10.12211/2096-8280.2021-014

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Production of sesquiterpenoids α-neoclovene and β-caryophyllene by engineered Saccharomyces cerevisiae

Research Article | 更新时间：2026-03-18

- Production of sesquiterpenoids α-neoclovene and β-caryophyllene by engineered Saccharomyces cerevisiae
- Synthetic Biology Journal Vol. 2, Issue 5, Pages: 792-803(2021)
- 作者机构：
  
  1.中国科学院分子植物科学卓越创新中心，合成生物学重点实验室，上海　200032
  2.中国科学院大学，北京　100049
  3.中国科学院深圳先进技术研究院，深圳合成生物学创新研究院，中国科学院定量工程生物学重点实验室，广东　深圳　518055
- 作者简介：
- 基金信息：
- DOI：10.12211/2096-8280.2021-014
  CLC： Q939.9;Q789
- Received：31 January 2021，
  
  Revised：2021-03-10，
  
  Published：31 October 2021
- 稿件说明：
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李晓东，杨成帅，王平平，严兴，周志华. 构建酿酒酵母细胞工厂从头合成倍半萜类化合物α-新丁香三环烯和β-石竹烯［J］. 合成生物学， 2021， 2（5）： 792-803

LI Xiaodong， YANG Chengshuai， WANG Pingping， YAN Xing， ZHOU Zhihua. Production of sesquiterpenoids α-neoclovene and β-caryophyllene by engineered Saccharomyces cerevisiae［J］. Synthetic Biology Journal， 2021， 2（5）： 792-803
李晓东，杨成帅，王平平，严兴，周志华. 构建酿酒酵母细胞工厂从头合成倍半萜类化合物α-新丁香三环烯和β-石竹烯［J］. 合成生物学， 2021， 2（5）： 792-803 DOI： 10.12211/2096-8280.2021-014.

LI Xiaodong， YANG Chengshuai， WANG Pingping， YAN Xing， ZHOU Zhihua. Production of sesquiterpenoids α-neoclovene and β-caryophyllene by engineered Saccharomyces cerevisiae［J］. Synthetic Biology Journal， 2021， 2（5）： 792-803 DOI： 10.12211/2096-8280.2021-014.

摘要

倍半萜类化合物（sesquiterpenoids）

-新丁香三环烯（

-neoclovene）和

-石竹烯（

-caryophyllene）都是人参挥发油中的主要组成成分，不仅具有重要的药用开发价值，而且在高能量密度生物能源的开发中也受到关注。然而，它们在人参和其他植物中含量均十分稀少且难以分离纯化，其开发与应用研究严重滞后。本研究利用天然产物合成生物学的方法，以BY4742为出发菌株构建了高产倍半萜类化合物前体FPP的通用底盘菌株SQTBY03。在此底盘菌株中异源表达了密码子优化的植物内生真菌

Hypoxylon

sp. EC38来源的多产物倍半萜合酶元件基因

ec38

和黄花蒿（

Artemisia annua

）来源的倍半萜合酶元件基因

QHS1

，分别构建了合成

-新丁香三环烯和

-石竹烯的两个倍半萜细胞工厂NCVBY01和CPLBY01。细胞工厂NCVBY01的主要倍半萜产物为

-新丁香三环烯（

39%），细胞工厂CPLBY01的主要产物为

-石竹烯（

96%）。它们的摇瓶发酵产量分别达到25.8 mg/L和250.4 mg/L。通过在1.3 L发酵罐中进行批次补料发酵，细胞工厂NCVBY01合成

-新丁香三环烯的产量达到了487.1 mg/L，为目前首次在微生物细胞工厂中合成

-新丁香三环烯；CPLBY01合成

-石竹烯的产量达到2949.1 mg/L，为目前报道的最高水平。上述研究成果为

-新丁香三环烯和

-石竹烯的规模化制备提供新的途径。

Abstract

Sesquiterpenoids

-neoclovene and

-caryophyllene are major components in volatile oils from

Panax ginseng

which have been demonstrated to play important roles in antibacteria

antitumor and cardiovascular protection. Moreover

they have attracted attentions for potential use as biofuels with high-energy-density. However

the industrial production of

-neoclovene and

-caryophyllene as well as other sesquiterpenoids are mainly relied on extraction from plant materials

which is too costly for applications at a large scale. Currently

this challenge could be addressed by advances in synthetic biology for natural product biosynthesis. Through hetero

logously assembling and integrating of their biosynthetic pathways into microbial chassis cells

targeted natural compounds from plants could be produced by microbial fermentation in a sustainable

low-cost and large-scale way. In this study

by comparing the production potential of sesquiterpenes between different

Saccharomyces cerevisiae

strains

and followed by enhancing the endogenous mevalonate pathway

a yeast sesquiterpene chassis strain (SQTBY03) with an increase of 458 times in farnesyl pyrophosphate production was constructed. Then by inserting the codon-optimized sesquiterpene synthase gene

ec38-cs

from the endophytic fungi

Hypoxylon

sp. EC38 and the codon-optimized caryophyllene synthase gene

QHS1

from

Artemisia annua

into SQTBY03

respectively

we built yeast cell factories NCVBY01 and CPLBY01 for

de novo

production of

-neoclovene and

-caryophyllene at their titers of 25.8 mg/L and 250.4 mg/L

respectively

in shake flasks. Furthermore

fed-batch fermentation using NCVBY01 and CPLBY01 resulted in the

de novo

production of 487.1 mg/L

-neoclovene and 2949.1 mg/L

-caryophyllene from glucose. It is also possible to further chemically catalyze

-caryophyllene to produce

-neoclovene. Our work provides strategies for the sustainable production of

-neoclovene and

-caryophyllene from glucose through microbial fermentation

which would benefit their applications as medicine and other functional products. In addition

our yeast chassis for the sesquiterpene production could offer a platform for the sustainable production of other valuable sesquiterpenoids

via

synthetic biology approach.

关键词

Keywords

references

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Related Institution

Key Laboratory of Synthetic Biology， Center for Excellence in Molecular Plant Sciences， Chinese Academy of Sciences

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⁰

Production of sesquiterpenoids α-neoclovene and β-caryophyllene by engineered Saccharomyces cerevisiae

DOI：10.12211/2096-8280.2021-014

摘要

Abstract

关键词

Keywords

references