PLANTA
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2021
·Vol. 253
·期 5
·1-8
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IF 4.116
Fu Jinqiu,Xu Wenbo,Huang Wei,Wang Bipeng,Li Shengchun,Zhang Jiang,Chang Ling
Main conclusion Importation of taxadiene synthase into chloroplasts is important for the efficient heterologous production of taxadiene. Abstract Taxadiene, the first committed precursor to taxol, is synthesized from geranylgeranyl pyrophosphate (GGPP) by action of taxadiene synthase (TS). Heterologous production of taxadiene could potentially rely on both cytosolic mevalonic acid (MVA) pathway and the plastidic methylerythritol phosphate (MEP) pathway. We suggest the compartmentalized engineering in chloroplast as an efficient approach for taxadiene production. In this study, we directly introduced the TS gene from Taxus brevifolia into the tobacco chloroplast genome and found that the transplastomic plants accumulated a low content of taxadiene, ~ 5.6 μg/g dry weight (DW). Moreover, we tried a combination of MEP and MVA pathways for taxadiene synthesis by nuclear transformation with a truncated version of TS (without encoding a transit peptide) into the transplastomic plants. However, this did not further improve the taxadiene production. In contrast, we found that taxadiene could be produced up to 87.8 μg/g DW in leaves of transgenic plants expressing TS with a chloroplast transit peptide, which was significantly higher than that in leaves of transplastomic plants. Thus, this study highlights the importance of TS importation into chloroplast for production of taxadiene.
研究领域:
植物学
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代谢工程
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生物化学
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PubMed
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ACS Synthetic Biology
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2025
·
IF 3.9
Baodong Hu,Jingwen Zhou,Jianghua Li,Jian Chen,Guocheng Du,Fang Zhong,Yucheng Zhao,Xinrui Zhao
Marmesin, a plant dihydrofuranocoumarin, is an important intermediate in the synthesis of linear furanocoumarins and exhibits a variety of pharmacological activities. However, due to the lack of efficient prenyltransferases, the incompatibility of redox partners for P450 enzymes, and the insufficient supply of precursor (DMAPP), the microbial synthesis of marmesin remained at an extremely low level. Here, we report the efficient biosynthesis of marmesin in Escherichia coli by screening the robust 6-prenyltransferase PpPT1 and marmesin synthase PpDCΔ2–29 from Peucedanum praeruptorum. Next, the activities of PpPT1 and PpDCΔ2–29 were enhanced using fusion protein tags and redox partner engineering, respectively. In addition, the synthesis of marmesin was further improved by strengthening the methylerythritol phosphate (MEP) pathway to increase the availability of DMAPP and by optimizing the modular pathway in the engineered strain. Finally, the titer of marmesin reached 203.69 mg L–1 in the fed-batch fermentation with a molar conversion rate of umbelliferone of 81.4%, which is the highest titer for marmesin production using engineered microorganisms. The applied strategy and marmesin-producing strain constructed in this study lay the foundation for the green production of valuable complex furanocoumarins.
研究领域:
生物化学
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合成生物学
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药理学
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AFRICAN JOURNAL OF BIOTECHNOLOGY
·
2017
·Vol. 16
·期 50
·2307-2316
·
IF 0
Haiyuan Liu,Shu Han,Liping Xie,Jie Pan,Wei Zhang,Guihua Gong,Youjia Hu
2-C-Methyl-D-erythritol-4-phosphate (MEP) pathway has been extensively employed for terpenoids biosynthesis in Escherichia coli. In this study, to obtain key-enzymes of MEP pathway for squalene production, overexpression of different combination of MEP pathway genes were compared. Squalene production in strain YSS12 with overexpressed dxs, idi and ispA of MEP pathway from E. coli was improved by 71-fold when compared with strain YSS3 which only contained double copy SQS. Analysis of transcriptional levels of MEP pathway genes in engineering strains showed that different squalene production can be attributed to changed transcriptional levels of co-overexpressed genes dxs, idi, ispG and ispA in engineering strains. Furthermore, different E. coli expression hosts were compared for squalene production, among which BL21(DE3) was the best squalene producer. These results illustrate that dxs, idi and ispA of the MEP pathway from E. coli were key-enzymes for squalene production in E. coli. These key-enzymes of MEP pathway could also be applied to other terpenoids production in E. coli.
研究领域:
微生物学
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代谢工程
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生物技术
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PubMed
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2025
·
IF 4
Zili Yang,Xin Chen,Liru Zheng,Qinping Hu,Mengwei Bu,Jiang Duan,Aidong Zhang
Inhibitor design based on the natural product fosmidomycin (FOS) as a lead structure is a promising strategy in search of herbicidally active compounds that may target the DXR enzyme in the MEP pathway. Herein, two metal ion chelating groups, aminol and amide, were used to replace the hydroxamate moiety of FOS, and two types of non-hydroxamate FOS analogs, namely aminol- ( 5a ∼ n and 6a ∼ o ) and phenylamide-containing ( 10a ∼ v ) phosphonic acids were designed and synthesized by establishing facile routes involving the Michaelis-Arbuzov reaction, epoxide ring-opening, acrylamidation, Michael addition, and phosphonate hydrolysis. In the pre-emergency bioactivity screening several compounds demonstrated herbicidal activities superior to FOS, including 5c , 5 h , 10a , 10d , 10e , etc. In particular, 10e showed an 18.7-fold inhibition activity against Arabidopsis thaliana and 16.1- and 10.8-fold activities against the root and stalk of Echinochloa crus-galli , respectively, in comparison with that of FOS. In the post-emergency assay, 10e also displayed 1.9- and 2.1-fold inhibition activities against E. crus-galli and Amaranthus retroflexus . Enzyme inhibition assay revealed the inhibition activities of some compounds to the DXR enzyme far below that of FOS, while the plant growth rescue failed from the inhibition by 5c and 10e by adding dimethylallyl pyrophosphate (DMAPP), a downstream product of the MEP pathway, both suggesting that some targets instead of the DXR enzyme might be involved in the inhibition by the active compounds. Nevertheless, this work demonstrated that using bioisosteric replacement based on the lead structure FOS is an effective strategy for developing highly herbicidal active compounds.
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INDUSTRIAL CROPS AND PRODUCTS
·
2021
·
IF 5.645
Zhengfeng Tian,Qingyun Luo,Zhaojiang Zuo
Plant terpenes are the main functional agents in forest bathing and have great value in several industries. To uncover the seasonal variation of Cinnamomum camphora monoterpene emission and the difference among the chemotypes for promoting the development of forest bathing and exploitation of monoterpenes using the species, the emission of monoterpenes and related gene expression in the camphor chemotype, eucalyptol chemotype, linalool chemotype and borneol chemotype of C. camphora were analyzed in May (spring), July (summer) and November (autumn), and the stored monoterpenes and storage structure were investigated. Eleocytes were the storage structure of monoterpenes in C. camphora , which widely existed in the leaf palisade tissue and spongy tissue, as well as around the glands and vascular bundles. The 4 chemotypes of C. camphora released different monoterpene components which mainly derived from the immediate emission after synthesis. Camphor, eucalyptol, linalool and endo-borneol showed the uppermost emission rate and were the typical monoterpenes in the corresponding chemotype, respectively, due to the high expression of the genes encoding bornyl diphosphate synthase ( E5.5.1.8 ), linalool synthase ( TSP14 ) and (+)-neomenthol dehydrogenase ( E1.1.1.208 ) for the biosynthesis of camphor, linalool and endo-borneol, respectively, and only expression of the gene encoding 1,8-cineole synthase ( TSP-cin ) for eucalyptol biosynthesis. Among the 4 chemotypes, the linalool chemotype showed the highest emission rate of monoterpenes in the same month (without significant difference with the eucalyptol chemotype in November), due to the high expression of the genes ( dxs , dxr , ispF , gcpE , and GGPS ) in methylerythritol-4-phosphate (MEP) pathway, α-terpineol synthase gene ( E4.2.3.111 ) and TSP14 . Among the 3 months, temperature was the remarkably varied meteorological condition. The highest monoterpene emission rate from the 4 chemotypes of C. camphora was detected in July, which should result from the up-regulated expression of related genes in monoterpene biosynthesis and improved vaporization and diffusion of stored monoterpenes under high temperature, with the former providing a major contribution.
研究领域:
植物学
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生物化学
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环境科学
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2016
·
IF 2.857
Min Shi,Xiuqin Luo,Guanhua Ju,Leilei Li,Shengxiong Huang,Tong Zhang,Huizhong Wang,Guoyin Kai
Tanshinones are health-promoting diterpenoids found in Salvia miltiorrhiza and have wide applications. Here, SmGGPPS (geranylgeranyl diphosphate synthase) and SmDXSII (1-deoxy-d-xylulose-5-phosphate synthase) were introduced into hairy roots of S. miltiorrhiza. Overexpression of SmGGPPS and SmDXSII in hairy roots produces higher levels of tanshinone than control and single-gene transformed lines; tanshinone production in the double-gene transformed line GDII10 reached 12.93 mg/g dry weight, which is the highest tanshinone content that has been achieved through genetic engineering. Furthermore, transgenic hairy root lines showed higher antioxidant and antitumor activities than control lines. In addition, contents of chlorophylls, carotenoids, indoleacetic acid, and gibberellins were significantly elevated in transgenic Arabidopsis thaliana plants. These results demonstrate a promising method to improve the production of diterpenoids including tanshinone as well as other natural plastid-derived isoprenoids in plants by genetic manipulation of the 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway.
研究领域:
植物生物技术
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代谢工程
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植物化学
·
基因工程
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ACS Synthetic Biology
·
2025
·
IF 3.9
Jerome R. Lon,Xuemei Zhao,Gulkiz Mamatrixat,Zhoukang Zhuang,Zhehao Jin,Tao Yu,Jufang Wang,Hongting Tang
Terpenoids are a diverse class of compounds with significant application potential. While prokaryotic bacteria synthesize terpenoids via the methylerythritol-4-phosphate (MEP) pathway, fungi utilize the mevalonate (MVA) pathway. The MVA pathway has been widely employed for efficient terpenoid production in bacteria such as Escherichia coli, but the MEP pathway performs poorly for biosynthesis in yeast. In this study, we constructed a compartmentalized MEP pathway to enhance monoterpenoid production in Saccharomyces cerevisiae. By introducing a geraniol synthase, we initially achieved the production of geraniol from glucose. Further effective incorporation of a cytosolic MEP pathway with nine enzymes increased geraniol production by 174.5%. However, this also significantly inhibited cell growth. Overexpression analysis revealed that flavodoxin and flavodoxin reductase were major contributors to growth inhibition, which could also be a factor limiting the application of the MEP pathway. To address these issues, we employed peroxisomal compartmentalization to isolate the MEP pathway from cytosolic metabolism. This strategy alleviated growth inhibition and improved geraniol production by 93.18% compared to that of cytosolic expression. Through additional metabolic engineering, we optimized peroxisomal geraniol production, achieving a yield of 30.64 mg/L. Our findings demonstrate the potential of compartmentalized MEP pathway expression as a viable approach for enhancing terpenoid biosynthesis in yeast, offering valuable insights for future metabolic engineering efforts.
研究领域:
合成生物学
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代谢工程
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微生物生物技术
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系统生物学
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生物制造
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PubMed
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GENE
·
2022
·
IF 3.913
Xiaohong Chen,Yueya Zhang,Haifeng Yan,Meiyun Niu,Yuping Xiong,Xinhua Zhang,Yuan Li,Jaime A. Teixeira da Silva,Guohua Ma
The commercial value of Santalum album L. lies in its aromatic heartwood and essential oil. Sesquiterpenes are the main components of sandal essential oil, and these are synthesized through the plant’s mevalonate (MVA) and methylerythritol phosphate (MEP) pathways. In this study, the first key rate-limiting enzyme, 1-deoxy- d -xylulose-5-phosphate synthase (SaDXS), was investigated to provide a theoretical molecular basis for the sandalwood MEP sesquiterpene biosynthetic pathway. The biofunctions of SaDXS were also analyzed. SaDXS promoters were successfully cloned from a seven-year-old S. album tree. SaDXS1A/1B promoter activity was verified by a β-glucuronidase (GUS) assay and by analyzing cis -acting elements of the promoters, which carried light- and methyl jasmonate (MeJA)-responsive signals. In an experiment involving yellow S. album seedlings, exposure to light upregulated SaDXS1A/1B expression and increased chlorophyll and carotenoid contents when overexpressed in Arabidopsis thaliana . Analysis of the expression of SaDXS1A/1B and SaSSy , key genes of santalol biosynthesis, revealed SaDXS1A expression in all tissues whereas SaDXS1B was expressed in tissues that contained photosynthetic pigments, such as stems, leaves and flowers. Sandal seedlings exogenously treated with two hormones, MeJA and ethylene, revealed similar expression patterns for SaDXS1A/1B and SaSSy . Sandal seedlings were treated with an inhibitor of DXS, clomazone, but showed no significant changes in the contents of α-santalene, β-santalene and α-santalol between treatment and control groups. These results suggest that SaDXS1A/1B play a role in the synthesis of sandalwood sesquiterpenes, providing carbon for downstream secondary metabolites. SaDXS1A/1B also play a role in the biosynthesis of chlorophyll, carotenoids, and primary metabolites.
研究领域:
植物生物技术
·
分子生物学
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生物化学
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植物化学
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PubMed
高影响因子
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