The colon's length increased significantly after receiving anemoside B4 (P<0.001), while the high-dose anemoside B4 group showed a decrease in the number of tumors (P<0.005). Analysis of the spatial metabolome showed anemoside B4 decreasing the quantity of fatty acids, their derivatives, carnitine, and phospholipids in colon tumor tissue. In parallel, anemoside B4 was observed to downregulate the expression of FASN, ACC, SCD-1, PPAR, ACOX, UCP-2, and CPT-1 in the colon, reaching statistically significant levels of suppression (P<0.005, P<0.001, P<0.0001). This study's findings suggest that anemoside B4 might restrain CAC through a regulatory effect on the reprogramming of fatty acid metabolism.
Pogostemon cablin's volatile oil, a complex mixture of various compounds, notably contains the sesquiterpenoid patchoulol, which is regarded as the primary contributor to its valuable pharmacological characteristics and aromatic profile, encompassing antibacterial, antitumor, antioxidant, and other biological functionalities. Patchoulol and its essential oil mixtures are presently in high demand across the world, but the traditional approach of plant extraction has significant drawbacks, including the squandering of land resources and the introduction of pollution into the environment. Subsequently, the development of a more economical and efficient technique for producing patchoulol is imperative. With the aim of increasing patchouli production methods and creating heterologous patchoulol synthesis in Saccharomyces cerevisiae, the patchoulol synthase (PS) gene from P. cablin underwent codon optimization and was placed under the control of the inducible, potent GAL1 promoter for introduction into the yeast platform strain YTT-T5. This resulted in strain PS00, which exhibits the production of 4003 mg/L patchoulol. Through the utilization of protein fusion methods, this study aimed to improve conversion rates. The fusion of the SmFPS gene from Salvia miltiorrhiza with the PS gene substantially increased patchoulol production, yielding a concentration of 100974 mg/L—a 25-fold elevation. Further refinement of the fusion gene's copy number significantly increased patchoulol output by 90%, reaching a concentration of 1911327 milligrams per liter. The strain, cultivated in a high-density fermentation system, showed improved patchouli yield, reaching 21 grams per liter, the highest yield seen to date thanks to an optimized fermentation process. The green production of patchoulol finds a crucial foundation in this investigation.
As an important economic tree species, Cinnamomum camphora plays a key role in China's economy. The volatile oil's key components in C. camphora leaves led to the classification of five chemotypes: borneol-type, camphor-type, linalool-type, cineole-type, and nerolidol-type. Terpene synthases (TPS) are the primary enzymes responsible for the creation of these compounds. Although research has identified several critical enzyme genes, the intricate biosynthetic pathway leading to (+)-borneol, the compound of most economic worth, has yet to be detailed. Transcriptome analysis of four chemically distinct leaves led to the cloning of nine terpenoid synthase genes, designated CcTPS1 to CcTPS9, in this investigation. The recombinant protein, induced within Escherichia coli, proceeded to use geranyl pyrophosphate (GPP) and farnesyl pyrophosphate (FPP) as substrates, respectively, in enzymatic reactions. CcTPS1 and CcTPS9 catalyze the transformation of GPP into bornyl pyrophosphate, which is then hydrolyzed by phosphohydrolase to produce (+)-borneol. The proportion of (+)-borneol generated is 0.04% from CcTPS1 and 8.93% from CcTPS9. Gpp is converted to linalool by both CcTPS3 and CcTPS6, and CcTPS6 further reacts with FPP to form nerolidol. The reaction between CcTPS8 and GPP yielded 18-cineol, a product present at 3071%. The nine terpene synthases collectively produced nine monoterpenes and six sesquiterpenes. Through this study, the key enzyme genes responsible for borneol biosynthesis in C. camphora have been identified for the first time, enabling a more thorough understanding of chemical type formation mechanisms and facilitating the creation of high-yield borneol varieties using bioengineering approaches.
Tanshinones, one of the key effective components present in Salvia miltiorrhiza, are important in the management of cardiovascular diseases. The production of tanshinones by microbial heterogony will give us a substantial source of ingredients for making traditional Chinese medicine (TCM) preparations of *Salvia miltiorrhiza*, consequently decreasing extraction costs and relieving the strain on clinical medication. The biosynthetic pathway of tanshinones involves a diverse array of P450 enzymes, with the high-efficiency catalytic element serving as a crucial foundation for their microbial production. Komeda diabetes-prone (KDP) rat Protein modification in CYP76AK1, a key P450-C20 hydroxylase within the tanshinone pathway, was investigated during this study. Utilizing the protein modeling methodologies SWISS-MODEL, Robetta, and AlphaFold2, the protein model was scrutinized to obtain a dependable protein structure. To design the mutant protein semi-rationally, molecular docking and homologous alignment procedures were undertaken. Molecular docking analysis revealed the key amino acid sites in CYP76AK1 that govern its oxidation capabilities. The function of the mutations obtained was investigated using a yeast expression system. Among these mutations, CYP76AK1 mutations exhibiting continuous oxidation of 11-hydroxysugiol were identified. A study of four key amino acid sites responsible for oxidation activity was undertaken, and the validity of three protein modeling techniques was examined in light of the resulting mutations. First reported herein are the effective protein modification sites of CYP76AK1, providing a catalytic element for diversified oxidation activities at the C20 position. This study's findings are instrumental for advancing tanshinone synthetic biology and establishing a framework for exploring the continuous oxidation mechanism of P450-C20 modification.
Heterologous biomimetic synthesis, a novel strategy in acquiring the active compounds of traditional Chinese medicine (TCM), exhibits significant promise for the protection and development of these resources. By replicating the synthesis of active compounds from medicinal plants and animals within biomimetic microbial cells, synthetic biology enables the scientific design and systematic reconstruction of key enzymes, thereby optimizing the heterologous production of these compounds by microorganisms. Target product acquisition, accomplished through this method, ensures efficient and environmentally responsible practices, driving large-scale industrial output and ultimately supporting the sustainable production of scarce Traditional Chinese Medicine resources. Beyond its core function, the method plays a significant role in agricultural industrialization, and introduces a new strategy for promoting green and sustainable TCM resource development. A comprehensive review of recent progress in heterologous biomimetic synthesis for traditional Chinese medicine active ingredients includes three focal points: the biosynthesis of terpenoids, flavonoids, phenylpropanoids, alkaloids, and other active components; the key obstacles and crucial aspects of heterologous biomimetic synthesis; and the application of biomimetic cells in the production of complex TCM mixtures. SARS-CoV2 virus infection This research project paved the way for using next-generation biotechnology and theories in the progress of Traditional Chinese Medicine.
The active compounds in traditional Chinese medicine (TCM) are crucial to the effectiveness of the therapy and to the creation of Dao-di herbs. Investigating the biosynthesis and regulatory mechanisms of these active compounds is crucial for understanding the formation process of Daodi herbs and developing active ingredient production strategies within Traditional Chinese Medicine (TCM) through the lens of synthetic biology. Thanks to the progression of omics technology, molecular biology, synthetic biology, artificial intelligence, and related areas, the analysis of biosynthetic pathways for active ingredients in Traditional Chinese Medicine is being expedited. Innovative methods and technologies have spurred the investigation of synthetic pathways of active compounds within Traditional Chinese Medicine (TCM), resulting in its emergence as a prominent area of study in molecular pharmacognosy. A considerable amount of progress has been made by researchers in the investigation of biosynthetic pathways for active components in traditional Chinese medicines like Panax ginseng, Salvia miltiorrhiza, Glycyrrhiza uralensis, and Tripterygium wilfordii. BLU-222 datasheet Current research methods for analyzing the biosynthetic functional genes of active ingredients found in Traditional Chinese Medicine were systematically evaluated in this paper, focusing on the identification of gene elements from multi-omics data and the experimental confirmation of these genes' functions in plant systems, encompassing both in vitro and in vivo analyses using candidate genes as targets. The paper, moreover, encapsulated the novel technologies and techniques, such as high-throughput screening, molecular probes, genome-wide association studies, cell-free systems, and computer simulations for screening, to provide a detailed reference on the study of biosynthetic pathways of active ingredients in Traditional Chinese Medicine.
Mutations in the inactive rhomboid 2 (iRhom2/iR2), encoded by the Rhbdf2 gene, are responsible for the rare familial disorder tylosis with esophageal cancer (TOC). iR2 and iRhom1 (or iR1, encoded by Rhbdf1) are essential regulators for the membrane-anchored metalloprotease ADAM17, which is crucial for activating EGFR ligands and releasing pro-inflammatory cytokines, including TNF (or TNF). Mice harboring a cytoplasmic deletion in iR2, which includes the TOC site, exhibit curly coats or bare skin (cub), contrasting with mice carrying a knock-in TOC mutation (toc), which manifest less severe alopecia and wavy fur. Amphiregulin (Areg) and Adam17 are implicated in the unusual skin and hair characteristics of iR2cub/cub and iR2toc/toc mice; the absence of one allele of either gene restores the fur's normal appearance.