ABSTRACT
Plant-derived aromatic natural products have important medicinal value and can be made into pharmaceutical and healthcare products with antibacterial, anti-inflammatory, analgesic, anti-oxidative, insecticidal and anthelmintic, expectorant and cough suppressant, tranquilizer and antitumor effects. However, the low content of aromatic natural products in plants and the difficulty and high costs in extraction and purification hampered its large-scale production and application. Recent advances in synthetic biology and metabolic engineering have enabled the tailor-made production of aromatic natural products using engineered microbial cell factories. This review summarizes the categories, the synthetic pathways, the key enzymes and the synthetic biology strategies for production of aromatic natural products, and discusses the challenges and opportunities in this area.
Subject(s)
Biological Products , Metabolic Engineering , Plants , Synthetic BiologyABSTRACT
Aromatic compounds make up a large part of fragrances and are traditionally produced by chemical synthesis and direct extraction from plants. Chemical synthesis depends on petroleum resources and has disadvantages such as causing environment pollutions and harsh reaction conditions. Due to the low content of aromatic compounds in plants and the low yield of direct extraction, plant extractions require large amounts of plant resources that occupy arable land. In recent years, with the development of metabolic engineering and synthetic biology, microbial synthesis of aromatic compounds from renewable resources has become a promising alternative approach to traditional methods. This review describes the research progress on the synthesis of aromatic fragrances by model microorganisms such as Escherichia coli or yeast, including the synthesis of vanillin through shikimic acid pathway and the synthesis of raspberry ketone through polyketide pathway. Moreover, this review highlights the elucidation of native biosynthesis pathways, the construction of synthetic pathways and metabolic regulation for the production of aromatic fragrances by microbial fermentation.
Subject(s)
Biosynthetic Pathways , Metabolic Engineering , Odorants , Shikimic Acid , Synthetic BiologyABSTRACT
Metabolic engineering has been developed for nearly 30 years since the early 1990s, and it has given a great impetus to microbial strain breeding and improvement. Aromatic chemicals are a variety of important chemicals that can be produced by microbial fermentation and are widely used in the pharmaceutical, food, feed, and material industry. Microbial cells can be engineered to accumulate a variety of useful aromatic chemicals in a targeted manner through rational engineering of the biosynthetic pathways of shikimate and the derived aromatic amino acids. This review summarizes the metabolic engineering strategies and biosynthetic pathways for the production of aromatic chemicals developed in the past 30 years, with the aim to provide a valuable reference and promote the research in this field.
Subject(s)
Biosynthetic Pathways , Fermentation , Metabolic Engineering , Shikimic AcidABSTRACT
Phytochemicals from Beet Root (Beta vulgaris) plant extract are traditionally used to cure Diarrhea. It is caused by Escherichia coli. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that caffeic acid can effectively deactivate the Shikimate dehydrogenase enzyme thereby interrupting the life cycle of the organism
ABSTRACT
Phytochemicals from Vaccinium corymbosum L.plant extract are traditionally used to cure diarrhea. It is caused by Escherichia coli. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that cafferic acid can effectively deactivate the shikimate dehydrogenase thereby interrupting the life cycle of the organism
ABSTRACT
ABSTRACT: Shikimic acid (SA) has witnessed a strong increase in recent years due to the increasing demand of the pharmaceutical and cosmetic industry. The SA is used as a precursor for the synthesis of oseltamivir phosphate (Tamiflu®), a potent viral inhibitor and is extracted from the plant Illicium verum Hook which has a limited availability. This article proposed the use of Urochloa plantaginea (Link.) webster and glyphosate, as an alternative source of SA. U. plantaginea plants with 3 - 4 tillers and 4 - 6 leaves were harvest at 3, 6, 9 and 12 days after application (DAT) of low rates of glyphosate. Samples were dried, extracted, analyzed by HPLC and LC-MS/MS. The maximum SA concentrations were observed at 6 days after glyphosate at 36 g.a.e.ha-1 was applied in plants of U. plantaginea with 4 to 6 leaves. The capability of this annual gramineae to produce elevated SA levels throughout the entire biomass affords its potential for a greater yield on a per hectare basis.
RESUMO: O interesse pelo ácido chiquímico (SA) tem apresentado um forte incremento nos últimos anos devido à crescente demanda da indústria farmacêutica e cosmética. O SA é utilizado como um precursor para a síntese do fosfato de oseltamivir (Tamiflu®), um potente inibidor viral. Este ácido é extraído principalmente da planta Illicium verum Hook. A disponibilidade desta planta é um fator limitante para o crescimento do mercado no futuro próximo. Este artigo propõe Urochloa plantaginea (Link.) webster tratada com sub doses de glifosato, como uma fonte alternativa de SA. Plantas de U. plantaginea com 3 - 4 perfilhos e 4 a 6 folhas foram tratadas com subdoses de glifosato e coletadas aos 0, 3, 6, 9 e 12 dias após sua aplicação (DAT). As amostras foram secas, extraídas e analisadas por HPLC e confirmadas por LC-MS/MS. As concentrações máximas de SA foram observadas aos seis dias após aplicação do glifosato a 36 g.e.a.ha-1 em plantas de U. plantaginea com 4 - 6 folhas. A capacidade anual dessa gramínea para produzir níveis elevados de SA em toda a biomassa, pode ser uma fonte economicamente viável de SA.
ABSTRACT
Objective::To clone p-coumaroyl quinate/shikimate 3' -hydroxylase gene from Lonicera macranthoides, and analyze its bioinformatics and expression patterns with chlorogenic acid content, in order to speculate the functions of LmC3H1 gene from L. macranthoides. Method::The full-length cDNA sequence of LmC3H1 gene was cloned by reverse trascription polymerase chain reaction(RT-PCR) and RACE techniques. The bioinformatics analysis of the gene sequence was carried out by using relevant software.Real-time fluorescence quantification PCR(Real-time PCR) and HPLC were used to determine relative expression of LmC3H1 and content of chlorogenic acid in stems, leaves and flowers of different flowering stages. Result::The LmC3H1 (GenBank: MN177695) gene was cloned, and the open reading frame (ORF) of it was 1 533 bp in length and encoded 510 amino acids. The molecular formula was C2618H4134N718O727S22, the relative molecular mass was 58 005.32, and the isoelectric point was 8.92.It was a hydrophilic protein located in the chloroplast with a transmembrane region LLLIPAVLFLISLVYPLI, and contained a conserved domain CYTOCHROME_P450(433-422 aa) in cytochrome P450.The results of Real-time PCR showed that LmC3H1 was expressed in different degrees in stems, leaves and different flowering stages of L. macranthoides. In the flower development stage, the relative expression of white bud stage was the highest, followed by flower buds and white flowering stage. The ratio of flower to stem and leaf was the highest, and the relative expression of flower was the highest. The HPLC results showed that the content of chlorogenic acid increased from greenish white to golden yellow in flowering stage and golden yellow flowering stage. Among the different organs, the flower had the highest chlorogenic acid, and the stem showed the lowest. Conclusion::The LmC3H1 gene of L. macranthoides is cloned, suggesting that LmC3H1 might be involved in the biosynthesis of L. macranthoides chlorogenic acid. This study provides a basis for further studying the functions of the gene and exploring the biosynthesis and regulation mechanism of L. macranthoides chlorogenic acid, while laying the foundation for the genetic improvement of L. macranthoides.
ABSTRACT
Objective To verify a method of calculation and prediction by screening the antibacterial components of classical Chinese medicinal herbs (TCM) for anti tuberculosis, and preliminarily evaluate the anti tuberculosis effect of the selected components. Methods The components database of Stemona, Bletilla striata and Chuanbei was established. Dock method was used to predict the anti tuberculosis components from the database, and then mycobacterium smegmatis, MIC and inhibition zone methods were used to evaluate the anti-tuberculosis effect of these predicted components. Results Three ingredients were selected. In vitro experiments also showed that the selected ingredients had the effect of anti-mycobacterium smegmatis by MIC and inhibition zone . Conclusions The virtual screening method could decrease consumption and increase the efficiency of finding anti-tuberculosis ingredients of TCM.
ABSTRACT
Shikimate kinase is a key protein of the shikimic pathway, which is essential for the survival of Mycobacterium tuberculosis. In this study, a screening assay for Mycobacterium tuberculosis shikimate ki-nase (MtSK) inhibitor was developed. A 120 000-compound library was screened by the enzyme assay and the phenotype screening using Mycobacterium smegmatis. A hit compound named IMB-T5297[(E)-3-(3-(3- chloro-5-methoxy-4-(prop-2-yn-1-yloxy)phenyl)acryloyl)-6-methyl-2H-pyran-2,4(3H)-dione] was identified to be a selective inhibitor of MtSK with a half maximal inhibitory concentration (IC50) value of 1.745 μg·mL-1, which also showed antibacterial activity. The interaction between compound and protein was analyzed by surface plasmon resonance (SPR) experiment, which showed the KD value was 2.151×10-5 mol·L-1. The binding model of MtSK and compound was simulated by the computer program. Five key amino acids in the binding pocket were indispensable site-directed mutated to verify the model. IMB-T5297 inhibited Mycobacterium tuberculosis H37Rv with a minimum inhibitory concentration (MIC) value of 49.723 μg·mL-1 and displayed low cytotoxicity to mammalian cells. In this study, IMB-T5297 was identified as a selective inhibitor of MtSK enzyme with anti-tuberculosis activity. With additional structural modification, the compound has a potential to become a novel anti-tuberculosis compound.
ABSTRACT
AIM@#To investigate the chemical constituents and their biological activities of the aerial parts of Euphorbia tibetica.@*METHOD@#Compounds were isolated and purified by various chromatographic methods, and their structures were elucidated through the use of extensive spectroscopic techniques including 2D-NMR, the structures of compounds 5 and 6 were confirmed by single-crystal X-ray crystallography. Bioactivities of all the isolated compounds were evaluated by the MTT method on A549 and anti-angiogenesis assay.@*RESULTS@#One new compound, methyl 4-epi-shikimate-3-O-gallate (1), together with twenty-three known constituents (2-24) were isolated from the aerial parts of E. tibetica.@*CONCLUSION@#Compound 1 is new, and the other compounds were isolated from this plant for the first time. Compounds 5-7, 9, 11, 17, 18 and 20 exhibited inhibitory effects on the growth of human lung cancer cell A549 and compounds 5, 7, 12, 13, 17 and 19 showed anti-angiogenic effects in a zebrafish model.
Subject(s)
Animals , Humans , Angiogenesis Inhibitors , Chemistry , Pharmacology , Cell Line , Cell Proliferation , Drugs, Chinese Herbal , Chemistry , Pharmacology , Euphorbia , Chemistry , Growth Inhibitors , Chemistry , Pharmacology , Molecular Structure , ZebrafishABSTRACT
Objective: To study the chemical constituents and their anti-oxidative activities of Kadsura coccinea. Methods: The compounds were isolated and purified by column chromatography. The structures were determined by means of physicochemical properties and spectroscopic analyses. The anti-oxidative activity was studied by DPPH methods. Results: Fourteen compounds were separated and identified as micrandilactone C (1), 3, 4-dimethoxy phenyl-α-D-glucopyranoside (2), salidroside (3), schisanhenol (4), protocatechuic acid (5), ethyl shikimate (6), shikimic acid (7), 5-O-β-D-glucopyranosylgentisic acid (8), syringic acid (9), 1, 2-benzenedicarboxylic acid-dibuty ester (10), benzyl-O-β-D-glucopyranoside (11), 4-O-β-D-glucopyranosylvanillic acid (12), syringaldehyde (13), and methyl shikimate (14). Compounds 5, 6, and 14 had the anti-oxidative activities. Conclusion: Compounds 2, 4, 6, and 8-14 are firstly isolated from K. coccinea. Compounds 5, 6, and 14 have the certain prospects in the development of natural anti-oxidative agents.
ABSTRACT
Com o objetivo de verificar o acúmulo de ácido chiquímico em plantas de laranja pêra (Citrus sinensis) num pomar comercial manejado com glifosato, um herbicida sistêmico de amplo espectro, foram coletadas amostras na Fazenda Jequitibá, tradicional no cultivo de citros, situada no Município de Santo Antônio de Posse, SP. O produtor aplicou de forma convencional Roundup® Original a 1.440 g.ha-1 de equivalente ácido (e.a.) do sal de isopropilamino de glifosato em 19/12/ 2006 na entrelinha de 15 plantas, deixando outras cinco como testemunha. A reaplicação de glifosato a 1.260 g.ha-1 de e.a. foi realizada em 2/4/2007. Em ambos os casos, imediatamente antes da aplicação e aos 3, 7, 10, 15, 20 e 35 dias após, foram coletadas 20 folhas de cada planta tanto da área tratada como da não tratada, analisando-se o teor de ácido chiquímico por cromatografia líquida de alta eficiência (CLAE) de forma isocrática após extração por micro-ondas. Os resultados mostraram não ocorrer acúmulo do ácido chiquímico nas plantas de laranja pêra, não havendo diferenças significativas nos teores deste composto entre o material proveniente da área tratada com glifosato e o daquela capinada manualmente.
In order to check the accumulation of shikimic acid in a traditional commercial grove of citrus "Pêra" cultivar (Citrus sinensis) managed for weed control with glyphosate, a systemic herbicide with wide spectrum, samples were collected at Fazenda Jequitibá, in Santo Antonio de Posse County, São Paulo State, Brazil. The producer applied the following treatments of Roundup Original® glyphosate at 1,440 g.ha-1 a.e. of the isopropylamine salt on 19 December 2006 between rows of 15 plants, leaving five others as control. The reapplication of glyphosate at 1,260 g ha-1 was done on 2 April 2007. In both cases, immediately before application and at 3, 7, 10, 15, 20 and 35 days thereafter, 20 leaves from each treated and untreated plants were collected for analysis of the content of shikimic acid by isocratic high performance liquid chromatography (HPLC) assisted with microwave. The results showed no significant differences in levels of shikimic acid between the material from the area treated with glyphosate and that weeded manually.
Subject(s)
Shikimic Acid/analysis , Citrus/parasitology , Herbicides , Chromatography, High Pressure LiquidABSTRACT
The 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase activity of Sclerotinia sclerotiorum is one of the multifunctional enzyme AROM activities, which catalyzes a reversible conversion of shikimate 3-phosphate (S3P) and phosphoenolpyruvate (PEP) to EPSP and inorganic phosphate, and is inhibited by the herbicide glyphosate (N-phosphonomethyl glycine). AROM protein has been purified from Sclerotinia sclerotiorum and the EPSP synthase has been analyzed. The results indicated that the optimal pH and temperature of EPSP synthase were 7.2 and 30℃ respectively. The activation energy of the heat-deactivated reaction of the enzyme was found to be 69.62 kJ/mol. Both of the substrates, S3P and PEP, were showed to inhibit the reaction rate when their concentrations exceeded 1 mmol/L and 2 mmol/L respectively. The Km of 140.98 μmol/L for PEP and 139.58 μmol/L for S3P were obtained by Dalziel equation which was a steadystate kinetic equation of the enzymatic reaction with the double substrates. The kinetic pattern of the enzyme was consistent with a sequential mechanism. Inhibition of the EPSP synthase reaction by glyphosate was competitive with respect to PEP, with the Ki 0. 32 μmol/L, and noncompetitive with regard to S3P. Activation by [ K+ ] was observed in the forward reaction. The Km (PEP) was lowered by increasing [ K+ ], while the Km (S3P) changed irregularly and the Ki (PEP) was enhanced.
ABSTRACT
In contrast to China where vegatation is predominantly herbaceous, vegetation in Brazil is commonly arboreous. This fact may explain why Chinese drugs are usually acetate derived, while actual and potential natural therapeutic agents from Brazil are mostly shikimate derived. Only relatively few compounds isolated from Brazilian plants have been submitted to adequate pharmacological testing .