Isolation of Taxol and Flavin-like fluorochrome from Endophytic Fungi of Mangifera indica

Scouting for novel and plant-derived biomolecules from endophytic microbial sources draws greater focus on the discovery of novel bioactive metabolites. With this rationale, we scouted the endophytic fungi for taxol, an anticancer diterpenoid and fluorescent biomolecules. In the present study, about 31 endophytic fungal isolates recovered from the Mangifera indica leaves were screened for taxol production in M1D medium. About five isolates were shortlisted based on the thin layer chromatographic analysis of the fungal extracts. Among them Colletotrichum sp. MIP-5 has been identified as a producer of fungal taxol based on UV, FTIR, TLC and HPLC analysis. The partially purified fungal taxol showed similar spectral and chromatographic features of commercially available paclitaxel. In addition to this, we also report the production of a fluorescent compound by Penicillium sp. MIP-3. The Flavin-like compound exhibited a bright greenish-yellow fluorescence with an emission maximum in the range of 505 – 545nm. GC-MS analysis showed the occurrence of Latia luciferin, primarily associated with the bioluminescence of freshwater limpet Latia neritoides. This is the first report of this compound from Penicillium sp. In addition, therapeutically active steroid (β-Sitosterol, Stigmasterol, Campesterol), quinones (Benzo[h]quinoline, 2,4-dimethyl-) and phloroglucinol (Aspidinol) derivatives were also identified from Penicillium sp. MIP-3 based on GC-MS analysis. These molecules could potentially be used in biological and pharmaceutical applications in future.

Taxus yunnanensis genome offers insights into gymnosperm phylogeny and taxol production

Taxol, a natural product derived from Taxus, is one of the most effective natural anticancer drugs and the biosynthetic pathway of Taxol is the basis of heterologous bio-production. Here, we report a high-quality genome assembly and annotation of Taxus yunnanensis based on 10.7 Gb sequences assembled into 12 chromosomes with contig N50 and scaffold N50 of 2.89 Mb and 966.80 Mb, respectively. Phylogenomic analyses show that T. yunnanensis is most closely related to Sequoiadendron giganteum among the sampled taxa, with an estimated divergence time of 133.4−213.0 MYA. As with most gymnosperms, and unlike most angiosperms, there is no evidence of a recent whole-genome duplication in T. yunnanensis. Repetitive sequences, especially long terminal repeat retrotransposons, are prevalent in the T. yunnanensis genome, contributing to its large genome size. We further integrated genomic and transcriptomic data to unveil clusters of genes involved in Taxol synthesis, located on the chromosome 12, while gene families encoding hydroxylase in the Taxol pathway exhibited significant expansion. Our study contributes to the further elucidation of gymnosperm relationships and the Taxol biosynthetic pathway.

Integrated system for paclitaxel production from endophytic fungus, Neopestalotiopsis clavispora ASU1 and subsequent extraction of chitosan from fungal biomass wastes

<p>Nowadays, breast cancer is considered to be one of the most prevalent diseases worldwide, this initiated interest and growing demand for taxol production in large quantities. The limited availability of traditional taxol production from the Pacific yew trees has encouraged research into the development of taxol production from alternative sources. Thus, the current study aimed to investigate the chemopreventive effect of paclitaxel derived from endophytic fungus Neopestalotiopsis clavispora ASU1. The endophytic fungus Neopestalotiopsis clavispora ASU1(KY624416) showed potent productivity of paclitaxel recording 100.6 µg/l which confirmed by HPLC, LC/Ms-Ms and FTIR analyses. In vitro, the extracted fungal taxol exerted a significant cytotoxic effect (P&lt; 0.05) at 300 nM, revealed that the increase in paclitaxel concentration induces increased cell death. Furthermore, the present study provides a promising approach for coupling paclitaxel production technology by endophytic fungus N. clavispora with chitosan production from residual fungal biomass resulting from taxol extraction and therefore improve the feasibility and commercialization of taxol production. The chitosan yield represented 5.94% of residual fungal dry biomass. Also, fungal chitosan was characterized for the degree of deacetylation (DD) (54.60%), FTIR spectroscopy, reducing power activity (0.263±0.051 mg/ml) may be attributed to hydroxyl groups (OH), and amine groups. These results confirmed that fungi are promising alternative sources for chitosan with superior physiochemical characteristics for food and medical prospective applications.</p>

Machine learning tools can now help with improving Taxol production in plant cell cultures

Chemotherapeutic intervention for cancer care is an important step. One of the most effective chemotherapy agents in use today is Paclitaxel (PTX), sold under the common name Taxol and Oxanol. Due to its ability to inhibit microtubule formation in cells, PTX is effective at all stages of the cancer and is FDA approved for treatment of many types of cancer (ovarian cancer, esophageal cancer, breast cancer, lung cancer, Kaposi sarcoma, cervical cancer, and pancreatic cancer). PTX is a plant alkaloid in the taxane family of compounds obtained from bark of the Pacific Yew tree (Taxus brevifolia) [1]. Adequate market supply of PTX has remained a challenge, as paclitaxel represents only a minor proportion of the total taxoid content of the Taxus species. Over the years, research into finding an alternate to cutting down Yew trees for PTX harvesting has been on the forefront. It is estimated that up to 60 trees may need to be harvested for the treatment of one patient.

Detection, Purification and Elucidation of Chemical Structure and Antiproliferative Activity of Taxol Produced by Penicillium chrysogenum

Penicillium chrysogenum has been reported as a potent taxol producer based on quantitative analysis by TLC and HPLC. The biosynthetic potency of taxol has been validated from PCR detection of rate-limiting genes of taxol synthesis such as taxadienesynthase and 10-de-acetylbaccatin III-O-acetyltransferase (DBAT), which catalyzes the immediate diterpenoid precursor of the taxol substance, as detected by PCR. Taxol production by P. chrysogenum was assessed by growing the fungus on different media. Potato dextrose broth (PDB) was shown to be the best medium for obtaining the higher amount of taxol (170 µg/L). A stepwise optimization of culture conditions necessary for production of higher amounts of taxol was investigated. The substance taxol was produced optimally after 18 d of incubation at 30 °C in PDB adjusted initially at pH 8.0 with shaking (120 rpm) (250 µg/L). The P. chrysogenum taxol was purified successfully by HPLC. Instrumental analyzes such as Fourier transform infrared spectroscopy (FTIR), ultraviolet (UV) spectroscopy, 1HNMR and 13C NMR approved the structural formula of taxol (C47H51NO14), as constructed by ChemDraw. The P. chrysogenum taxol showed promising anticancer activity.

Exploiting the Biosynthetic Potency of Taxol from Fungal Endophytes of Conifers Plants; Genome Mining and Metabolic Manipulation

Endophytic fungi have been considered as a repertoire for bioactive secondary metabolites with potential application in medicine, agriculture and food industry. The biosynthetic pathways by fungal endophytes raise the argument of acquisition of these machineries of such complex metabolites from the plant host. Diterpenoids “Taxol” is the most effective anticancer drug with highest annual sale, since its discovery in 1970 from the Pacific yew tree, Taxus brevifolia. However, the lower yield of Taxol from this natural source (bark of T. brevifolia), availability and vulnerability of this plant to unpredicted fluctuation with the ecological and environmental conditions are the challenges. Endophytic fungi from Taxus spp. opened a new avenue for industrial Taxol production due to their fast growth, cost effectiveness, independence on climatic changes, feasibility of genetic manipulation. However, the anticipation of endophytic fungi for industrial Taxol production has been challenged by the loss of its productivity, due to the metabolic reprograming of cells, downregulating the expression of its encoding genes with subculturing and storage. Thus, the objectives of this review were to (1) Nominate the endophytic fungal isolates with the Taxol producing potency from Taxaceae and Podocarpaceae; (2) Emphasize the different approaches such as molecular manipulation, cultural optimization, co-cultivation for enhancing the Taxol productivities; (3) Accentuate the genome mining of the rate-limiting enzymes for rapid screening the Taxol biosynthetic machinery; (4) Triggering the silenced rate-limiting genes and transcriptional factors to activates the biosynthetic gene cluster of Taxol.

The Research Progress of taxol in Taxus

Background: Taxus is a valuable woody species with important medicinal value. The bark of Taxus can produce taxol, a natural antineoplastic drug that is widely used in the treatment of breast, ovarian and lung cancers. However, the low content of taxol in the bark of Taxus can not meet the growing clinical demands, so the current research is on finding ways to increase taxol production. Objective: In this review, the research progress of taxol including the factors affecting the taxol content, biosynthesis pathway of taxol, production of taxol in vitro and the application of multi-omics approaches in Taxus as well as future research prospects will be discussed. Results and Conclusion: The taxol content is not only dependent on the species, age and tissues, but also affected by light, moisture levels, temperature, soil fertility and microbes. Most of the enzymes in taxol biosynthesis pathway have been identified and characterized. Total chemical synthesis, semi-synthesis, plant cell culture and biosynthesis in endophytic fungi have been explored to product taxol. Multi-omics have been used to study on Taxus and taxol. Further efforts in the identification of unknown enzymes in taxol biosynthesis pathway, establishment of the genetic transformation system in Taxus and the regulatory mechanism of taxol biosynthesis and Taxus cell growth will play a significant role in improving the yield of taxol in Taxus cells and plants.