JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2020
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IF 4.192
Ying Zhou,Rufang Deng,Xinlan Xu,Ziyin Yang
Linalool is abundant in tea leaves and contributes greatly to tea aroma. The two isomers of linalool, (R)-linalool and (S)-linalool, exist in tea leaves. Our study found that (R)-linalool was the minor isomer in nine of Camellia sinensis var. sinensis cultivars. The (R)-linalool synthase of tea plant CsRLIS was identified subsequently. It is a chloroplast-located protein and specifically catalyzes the formation of (R)-linalool in vitro and in vivo. CsRLIS was observed to be a stress-responsive gene and caused the accumulation of internal (R)-linalool during oolong tea manufacture, mechanical wounding, and insect attack. Further study demonstrated that the catalytic efficiency of CsRLIS was much lower than that of (S)-linalool synthase CsSLIS, which might explain the lower (R)-linalool proportion in C. sinensis var. sinensis cultivars. The relative expression levels of CsRLIS and CsSLIS may also affect the (R)-linalool proportions among C. sinensis var. sinensis cultivars. This information will help us understand differential distributions of chiral aroma compounds in tea.
研究领域:
植物学
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分子生物学
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芳香化合物代谢
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PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2019
·
IF 3.571
Bin Ma,Min Liu,Zhen-Hai Li,Xinyi Tao,Dong-Zhi Wei,Feng-Qing Wang
Patchoulol, a natural sesquiterpene compound, is widely used in perfumes and cosmetics. Several strategies were adopted to enhance patchoulol production in Saccharomyces cerevisiae: (i) farnesyl pyrophosphate (FPP) synthase and patchoulol synthase were fused to increase the utilization of FPP precursor; (ii) expression of the limiting genes of the mevalonate pathway was enhanced; (iii) squalene synthase was weakened by a glucose-inducible promoter of HXT1 (promoter for hexose transporter) to reduce metabolic flux from FPP to ergosterol; and (iv) farnesol biosynthesis was inhibited to decrease the consumption of FPP. Glucose was used to balance the trade-off between the competitive squalene and patchoulol pathways. The patchoulol production was 59.2 ± 0.7 mg/L in a shaken flask with a final production of 466.8 ± 12.3 mg/L (20.5 ± 0.5 mg/g dry cell weight) combined with fermentation optimization, which was 7.8-fold higher than the reported maximum production. The work significantly promoted the industrialization process of patchoulol production using biobased microbial platforms.
研究领域:
生物技术
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代谢工程
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生物过程工程
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PubMed
高影响因子
Scientific Reports
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2016
·Vol. 6
·期 1
·1-8
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IF 5.228
Su Ping,Tong Yuru,Cheng Qiqing,Hu Yating,Zhang Meng,Yang Jian,Teng Zhongqiu,Gao Wei,Huang Luqi
Salvia miltiorrhiza Bunge is highly valued in traditional Chinese medicine for its roots and rhizomes. Its bioactive diterpenoid tanshinones have been reported to have many pharmaceutical activities, including antibacterial, anti-inflammatory and anticancer properties. Previous studies found four different diterpenoid biosynthetic pathways from the universal diterpenoid precursor ( E , E , E )-geranylgeranyl diphosphate (GGPP) in S. miltiorrhiza . Here, we describe the functional characterization of ent -copalyl diphosphate synthase (SmCPS ent ), kaurene synthase (SmKS) and kaurene oxidase (SmKO) in the gibberellin (GA) biosynthetic pathway. SmCPS ent catalyzes the cyclization of GGPP to ent -copalyl diphosphate ( ent -CPP), which is converted to ent -kaurene by SmKS. Then, SmKO catalyzes the three-step oxidation of ent -kaurene to ent -kaurenoic acid. Our results show that the fused enzyme SmKS-SmCPS ent increases ent -kaurene production by several fold compared with separate expression of SmCPS ent and SmKS in yeast strains. In this study, we clarify the GA biosynthetic pathway from GGPP to ent -kaurenoic acid and provide a foundation for further characterization of the subsequent enzymes involved in this pathway. These insights may allow for better growth and the improved accumulation of bioactive tanshinones in S. miltiorrhiza through the regulation of the expression of these genes during developmental processes.
研究领域:
植物学
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药用植物研究
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分子生物学
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PubMed
高影响因子
TETRAHEDRON
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2022
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IF 2.388
Li Rao,Hu-Cheng Shi,Yi Zou
A new fungal nonribosomal peptide-polyketide hybrid synthase , namely CogA, was discovered by genome mining from fungus Cordyceps gunnii . Its ability to synthesize 4-hydroxyl 2-pyrrolidinone alkaloid ( 1 ) via nonenzyme-catalyzed N-1-C-2 closure was confirmed by heterologous expression of it in Aspergillus nidulans . Further engineered biosynthesis of the fluoro-substituted 2-pyrrolidinone alkaloid analogues ( 4 and 5 ) has also been achieved in vivo .
研究领域:
真菌学
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合成生物学
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生物化学
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高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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2018
·Vol. 102
·期 14
·6039-6046
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IF 3.34
Tigu Fitsum,Zhang Junli,Liu Guoxia,Cai Zhen,Li Yin
d -Pantothenic acid (vitamin B 5 ) has wide applications in the feed, food, chemical, and pharmaceutical industries. Its biological production routes which employ pantothenate synthetase (PS) as the key enzyme are attractive since they avoid the tedious and time-consuming optical resolution process. However, little data is available on the activity and kinetics of this enzyme, hampering the rational selection of an efficient enzyme for the biological production of d -pantothenic acid. In this study, six phylogenetically distant PS-encoding genes, from Escherichia coli , Corynebacterium glutamicum , Bacillus subtilis , Bacillus thuringiensis , Bacillus cereus , and Enterobacter cloacae , were expressed in E. coli . The PS from C. glutamicum exhibited a specific activity of 205.1 U/mg and a turnover number of 127.6 s -1 , which to our best knowledge are the highest values ever reported. The addition of substrates ( d -pantoic acid and β-alanine) to the E. coli strain harboring this enzyme during the early log phase of fermentation resulted in the production of 97.1 g/L of d -pantothenic acid within 32 h, corresponding to a conversion yield of 99.1% and a productivity of 3.0 g/L/h. To the best of our knowledge, this is the highest productivity reported to date.
研究领域:
生物化学
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微生物学
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工业生物技术
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高影响因子
INDUSTRIAL CROPS AND PRODUCTS
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2022
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IF 6.449
Shu Diao,Yini Zhang,Qifu Luan,Xianyin Ding,Jiaming Sun,Jingmin Jiang
Oleoresin terpenoids have been widely used in the food , pharmaceutical, and chemical industries. A thorough understanding of oleoresin terpenoid biosynthesis is important for the metabolic engineering of high-valued terpenoid products in plants and microbes and the improvement of oleoresin traits using molecular breeding strategies. TPS-d subfamily members, specific to conifers, determine the oleoresin terpenoid compound diversity. Here, we aimed to identify the TPS-d genes in slash pine ( Pinus elliottii ) and characterize their function and expression patterns in population. Our study reveals two main results: (1) A total of 52 terpene synthase (TPS) candidate genes were identified based on long-read transcripts of slash pine, 12 of which were successfully cloned from the secondary developing xylem RNA. Two levopimaradiene/abietadiene synthases (LAS), one monofunctional isopimaradiene synthase (mISO1)-like, and two α-pinene synthases were identified by comparing with previously known TPS. Next, the two α-pinene synthases and one β-pinene synthase producing multiple products were validated through heterologous expression and in vitro enzyme activity assays, which showed different product profiles from previously identified TPS. (2) The expression status of the cloned TPS genes in 240 slash pine individuals were analyzed. Four exhibited very low expression levels, including those identical to copalyl diphosphate synthases, hemiterpene synthases, diterpene synthases without activity, and one obtained without soluble protein in the heterologous expression. The expression levels of eight TPS genes participating in slash pine oleoresin biosynthesis varied in different individuals and were highly correlated with multiple transcriptional regulators, namely, MYB and MYC. The expression of two α-pinene synthase genes was significantly higher than that of the other TPS genes. Some TPS gene expression levels were positively correlated with the corresponding oleoresin component content. Our results provide an essential reference for exploring the mechanisms of terpene biosynthesis and regulation.
研究领域:
植物生物化学
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分子生物学
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遗传学
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代谢工程
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高影响因子
Horticulture Research
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2025
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IF 8.5
Zhang Yiyang,Zhang Yuhan,Zhang Andong,Tian Qiurui,Yang Bin,Wei Likun,Wu Wei,Zhu Ting,Zhou Zhiwei,Wang Jiaqi,Liu Zhibin,Tang Wei,Xiao Haijun,Liu Mingchun,Li Tao,Sun Qun
Epiphyllum oxypetalum, a renowned ornamental species in Cactaceae, releases attractive fragrance during its infrequent, transient and nocturnal blooms. However, the floral fragrance composition and biosynthesis remain largely unexplored. Employing volatilomics, transcriptomics and biochemistry, we systematically characterized the composition, emission dynamics and biosynthesis of the floral scent of E. oxypetalum. The floral scent composition of E. oxypetalum was highly dynamic. Starting after 20:00 local time, volatile emission increased 200-fold within six hours. At full bloom, geraniol accounted for 72.54% of the total emission, followed by benzyl alcohol (12.96%) and methyl salicylate (3.75%). These scents predominantly originated from petals and sepals. Transcriptomic analysis and inhibition assays using pathway-specific inhibitors revealed that the mevalonate pathway was the precursor source for geraniol biosynthesis. Functionally characterized cytosol localizing geraniol synthase EoTPSa1 was the key enzyme responsible for geraniol biosynthesis. Together, these findings pinpointed a cytosolic biosynthetic route for the major scent volatile geraniol in E. oxypetalum. Our study provides new insights into the emission dynamics and biosynthesis of E. oxypetalum floral scents. In particular, we demonstrated a distinctive mevalonate pathway-based geraniol biosynthetic pathway, which may hold potential for the development of novel perfume products.
研究领域:
植物学
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生物化学
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PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Haichun Zeng,Beilin Meng,Jiatong Zeng,Li Rao
Sesquiterpenes, a structurally diverse class of natural products, play pivotal roles in the pharmaceutical, fragrance, and flavor industries. In this study, we first identified and functionally characterized a multifunctional sesquiterpene synthase, ChTPS6, from the plant-pathogenic fungus Cochliobolus heterostrophus. When incubated with FPP, ChTPS6 primarily catalyzed the formation of cubebol as the main product, which is widely used as a sustained cooling and refreshing agent in the food industry. Remarkably, ChTPS6 exhibited substrate promiscuity, efficiently utilizing GPP and GGPP as alternative substrates. Moreover, through protein modeling and site-directed mutagenesis, we pinpointed key amino acid residues essential for the catalytic activity of ChTPS6, and successfully re-engineered the enzyme to alter its product specificity. These findings provide novel insights into the catalytic mechanism and functional versatility of ChTPS6 while broadening its potential applications in the biosynthesis of valuable sesquiterpenes.
研究领域:
生物化学
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分子生物学
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真菌学
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PubMed
高影响因子
BIOCHEMISTRY
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2016
·
IF 2.876
Zhenqiu Li,Ruiping Gao,Qinggang Hao,Huifang Zhao,Longbin Cheng,Fang He,Li Liu,Xiuhua Liu,Wayne K. W. Chou,Huajie Zhu,David E. Cane
The T296V mutant of amorpha-4,11-diene synthase catalyzes the abortive conversion of the natural substrate (E,E)-farnesyl diphosphate mainly into the acyclic product (E)-β-farnesene (88%) instead of the natural bicyclic sesquiterpene amorphadiene (7%). Incubation of the T296V mutant with (3R,6E)-nerolidyl diphosphate resulted in cyclization to amorphadiene. Analysis of additional mutants of amino acid residue 296 and in vitro assays with the intermediate analogue (2Z,6E)-farnesyl diphosphate as well as (3S,6E)-nerolidyl diphosphate demonstrated that the T296V mutant can no longer catalyze the allylic rearrangement of farnesyl diphosphate to the normal intermediate (3R,6E)-nerolidyl diphosphate, while retaining the ability to cyclize (3R,6E)-nerolidyl diphosphate to amorphadiene. The T296A mutant predominantly retained amorphadiene synthase activity, indicating that neither the hydroxyl nor the methyl group of the Thr296 side chain is required for cyclase activity.
研究领域:
植物生物学
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分子生物学
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PubMed
高影响因子
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY
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2014
·Vol. 33
·期 2
·375-388
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IF 3.55
Huang Run-Yue,Chu Yong-Liang,Jiang Ze-Bo,Chen Xiu-Ming,Zhang Xian,Zeng Xing
Background/Aims: The effects of glycyrrhizin treatment in lung cancer remain undetermined, despite extensive studies of the anti-tumor activities of glycyrrhizin. Methods: Lung adenocarcinoma A549 and NCI-H23 cell lines were used in this study. Cell growth was examined by MTS assays, while apoptosis and cell cycle were determined by flow cytometric analysis. Both real-time PCR and western blotting were used to examine the expression levels of thromboxane synthase (TxAS), and TxAS activity was measured using EIA detection of the biosynthesis of TxA2. TxAS was overexpressed in NCI-H23 cells by transfection with TxAS cDNA, while TxAS was inhibited by transfection with TxAS siRNA in A549 cells. For the mouse model of lung adenocarcinoma, the effects of glycyrrhizin on tumor growth were analyzed by western blot evaluation of TxAS, PTEN and survivin. TxAS activity was determined by EIA assay. Results: Glycyrrhizin suppressed cell growth in A549 cells, but not in NCI-H23 cells, by induction of apoptosis. TxAS was overexpressed in A549 cells, but the TxAS levels in NCI-H23 cells were minimal. Moreover, TxAS expression and activity were suppressed by glycyrrhizin. Glycyrrhizin had no additive effects with TxAS siRNA knockdown in suppressing A549 cell growth, whereas it completely suppressed cell growth of NCI-H23 cells transfected with TxAS cDNA. These results were further confirmed by the in vivo study. Conclusion: Our study suggests that the anti-tumor effect of glycyrrhizin in lung adenocarcinoma is, at least in part, TxAS-dependent. Therefore, glycyrrhizin is a promising anti-cancer agent for the treatment of lung adenocarcinoma
研究领域:
肿瘤学
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分子生物学
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药理学
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PubMed
高影响因子
PLANT PHYSIOLOGY AND BIOCHEMISTRY
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2022
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IF 5.437
Dahe Qiao,Mengsha Tang,Ling Jin,Xiaozeng Mi,Hongrong Chen,Junyan Zhu,Shengrui Liu,Chaoling Wei
Monoterpenes and sesquiterpenes are the most abundant volatiles in tea plants and have dual functions in aroma quality formation and defense responses in tea plants. Terpene synthases (TPS) are the key enzymes for the synthesis of terpenes in plants; however, the functions of most of them in tea plants are still unknown. In this study, six putative terpene biosynthesis gene clusters were identified from the tea plant genome. Then we cloned three new TPS-b subfamily genes, CsTPS08 , CsTPS10 and CsTPS58 . In vitro enzyme assays showed that CsTPS08 and CsTPS58 are two multiple-product terpene synthases, with the former synthesizing linalool as the main product, and β-myrcene, α-phellandrene, α-terpinolene, D-limonene, cis-β-ocimene, trans-β-ocimene and (4E,6Z)-allo-ocimene as minor products are also detected, while the latter catalyzing the formation of α-pinene and D-limonene using GPP as the substrate. No product of CsTPS10 was detected in the prokaryotic expression system , but geraniol production was detected when transiently expressed in tobacco leaves. CsTPS08 and CsTPS10 are two functional members of a monoterpene synthase gene cluster, which were significantly induced during both Ectropis oblique feeding and fresh leaf spreading treatments, suggesting that they have dual functions involved in tea plant pest defense and tea aroma quality regulation. In addition, the differences in their expression levels in different tea plant cultivars provide a possibility for the subsequent screening of tea plant resources with a specific aroma flavor. Our results deepen the understanding of terpenoid synthesis in tea plants.
研究领域:
植物学
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分子生物学
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基因组学
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高影响因子
Microbial Cell Factories
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2016
·Vol. 15
·期 1
·1-9
·
IF 3.744
Yang Jianming,Nie Qingjuan
Background Under aerobic conditions, acetic acid is the major byproduct produced by E. coli during the fermentation. And acetic acid is detrimental to cell growth as it destroys transmembrane pH gradients. Hence, how to reduce the production of acetic acid and how to utilize it as a feedstock are of intriguing interest. In this study, we provided an evidence to produce β-caryophyllene by the engineered E. coli using acetic acid as the only carbon source. Results Firstly, to construct the robust acetate-utilizing strain, acetyl-CoA synthases from three different sources were introduced and screened in the E. coli . Secondly, to establish the engineered strains converting acetic acid to β-caryophyllene, acetyl-CoA synthase (ACS), β-caryophyllene synthase (QHS1) and geranyl diphosphate synthase (GPPS2) were co-expressed in the E. coli cells. Thirdly, to further enhance β-caryophyllene production from acetic acid, the heterologous MVA pathway was introduced into the cells. What’s more, acetoacetyl-CoA synthase (AACS) was also expressed in the cells to increase the precursor acetoacetyl-CoA and accordingly resulted in the increase of β-caryophyllene. The final genetically modified strain, YJM67, could accumulate the production of biomass and β-caryophyllene up to 12.6 and 1.05 g/L during 72 h, respectively, with a specific productivity of 1.15 mg h −1 g −1 dry cells, and the conversion efficiency of acetic acid to β-caryophyllene (gram to gram) reached 2.1 %. The yield of β-caryophyllene on acetic acid of this strain also reached approximately 5.6 % of the theoretical yield. Conclusions In the present study, a novel biosynthetic pathway for β-caryophyllene has been investigated by means of conversion of acetic acid to β-caryophyllene using an engineered Escherichia coli . This was the first successful attempt in β-caryophyllene production by E. coli using acetic acid as the only carbon source. Therefore, we have provided a new metabolic engineering tool for β-caryophyllene synthesis.
研究领域:
生物技术
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代谢工程
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微生物学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）