Evaluation of spore inoculum and confirmation of pathway genetic blueprint of T13αH and DBAT from a Taxol-producing endophytic fungus

AbstractTaxol (paclitaxel), a plant-derived anticancer drug, has been among the most successful anticancer drugs of natural origin. Endophytic fungi have been proposed as a prominent alternative source for Taxol and its intermediate Baccatin III, however the very low yields remain a hinderance to their commercial utilization. Significant research efforts towards this end are underway globally. Here, we report the results on our earlier reported Taxol-producing endophytic fungus, Fusarium solani from the standpoint of spores as seed inoculum and media selection for enhanced Taxol and baccatin III yields. Spores produced on M1D medium with 94.76% viability were used for further media optimization for Taxol and Baccatin III production in five different liquid media under static and shaker condition at different cultivation days. Taxol and Baccatin III when quantified through competitive inhibition enzyme immunoassay (CIEIA), showed maximum production at 136.3 µg L−1 and 128.3 µg L−1, respectively in the modified flask basal broth (MFBB) under shaking condition. Further, two important genes of this pathway, namely taxane 13α-hydroxylase (T13αH) and 10-deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) have been identified in this fungus. These findings are hoped to assist in further manipulation and metabolic engineering of the parent F. solani strain towards the enhanced production of Taxol and baccatin III.

Download Full-text

Biochemical insights into the recombinant 10-deacetylbaccatin III-10-β-O-acetyltransferase enzyme from the Taxol-producing endophytic fungus Lasiodiplodia theobromae

FEMS Microbiology Letters ◽

10.1093/femsle/fnz072 ◽

2019 ◽

Vol 366 (7) ◽

Author(s):

Balendra Sah ◽

Kamalraj Subban ◽

Chelliah Jayabaskaran

Keyword(s):

Endophytic Fungi ◽

Endophytic Fungus ◽

Biosynthetic Pathway ◽

Gel Filtration ◽

Alternative Source ◽

Lasiodiplodia Theobromae ◽

Reading Frame ◽

Baccatin Iii ◽

Feasible Alternative ◽

Near Future

ABSTRACT 10-deacetylbaccatin III-10-β-O-acetyltransferase (DBAT) is a key rate-limiting enzyme of the Taxol biosynthetic pathway, which is uncharacterized in Taxol-producing endophytic fungi. Here, an open reading frame of DBAT was cloned from the Taxol-producing endophytic fungus Lasiodiplodia theobromae (LtDBAT). The LtDBAT enzyme was heterologously expressed and purified by the affinity and gel filtration chromatography methods. The molecular weight of the purified protein was 49 kDa and its identity was confirmed by western blot. The purified LtDBAT enzyme was capable of catalyzing 10-deacetylbaccatin III into baccatin III, as shown by liquid chromatography–mass spectroscopy. The mass spectra of baccatin III were identical to the authentic baccatin III. The LtDBAT enzyme was characterized and the kinetic parameters of catalysis were determined. In addition, localization of LtDBAT was performed by using confocal microscopy and the result showed that the enzyme was localized in lipid droplets. Together, this study provides biochemical insights into the fungal recombinant DBAT enzyme that is involved in the Taxol biosynthetic pathway. In the near future, engineering of the LtDBAT enzyme and the Taxol biosynthetic pathway in endophytic fungi could be an eco-friendly and economically feasible alternative source for production of Taxol and its precursors.

Download Full-text

Inhibition of cancer cell proliferation and apoptosis-inducing activity of fungal taxol and its precursor baccatin III purified from endophytic Fusarium solani

Cancer Cell International ◽

10.1186/1475-2867-13-105 ◽

2013 ◽

Vol 13 (1) ◽

pp. 105 ◽

Cited By ~ 16

Author(s):

Balabhadrapatruni VSK Chakravarthi ◽

Ramanathan Sujay ◽

Gini C Kuriakose ◽

Anjali A Karande ◽

Chelliah Jayabaskaran

Keyword(s):

Cell Proliferation ◽

Cancer Cell ◽

Fusarium Solani ◽

Cancer Cell Proliferation ◽

Baccatin Iii ◽

Proliferation And Apoptosis

Download Full-text

Culture media selection for mass production of Isaria fumosorosea and Isaria farinosa

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132010000400002 ◽

2010 ◽

Vol 53 (4) ◽

pp. 753-761 ◽

Cited By ~ 25

Author(s):

Gabriel Moura Mascarin ◽

Sérgio Batista Alves ◽

Rogério Biaggioni Lopes

Keyword(s):

Mass Production ◽

Culture Media ◽

Fungal Species ◽

Complete Medium ◽

Sugarcane Molasses ◽

Liquid Media ◽

Isaria Fumosorosea ◽

Isaria Farinosa ◽

Selection For ◽

Biphasic Fermentation

This work investigated the production of the fungi Isaria fumosorosea and Isaria farinosa in biphasic fermentation using agro-industrial products and residues. Combinations of natural liquid substrates, alternative to the complete medium and potato dextrose medium, were evaluated. The best liquid media were sugarcane molasses + rice broth, rice broth + yeast and sugarcane molasses + yeast + rice broth, which resulted in the highest viable propagule concentration. The molasses + rice broth medium was selected for the next phase of the study in which the production of both fungal isolates was evaluated in solid grain substrates. In solid-state fermentation, the best conidia production was achieved with the soybean meal and broken corn for I. farinosa, and whole rice and broken rice for I. fumosorosea. Results demonstrated that the two fungal species could be rapidly produced with higher yield of conidia on agro-industrial resources by using biphasic fermentation techniques.

Download Full-text

Study of the Effect of Nutrient Environment on the Biological Characteristics of Fusarium solani Clinical Corneal Isolates

Journal of Ophthalmology ◽

10.1155/2021/6666303 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Dalan Jing ◽

Yingyu Li ◽

Ziyuan Liu ◽

Chen Huang ◽

Pei Zhang ◽

...

Keyword(s):

Fusarium Solani ◽

Glucose Concentration ◽

Environmental Changes ◽

Biological Characteristics ◽

Clinical Isolates ◽

Reproductive Capacity ◽

Diabetic Patients ◽

Liquid Media ◽

Nutritional Conditions

Purpose. The biological characteristics of Fusarium solani clinical corneal isolates growing in different nutritional conditions in vitro were studied in order to find out the key point of pathogenicity. Methods. Five kinds of media with different glucose and nitrogen concentrations were prepared as the liquid and solid forms. The clinical isolates were as follows: 2 Fusarium solani strains. The clinical corneal isolates and the standard strains were inoculated in the solid and liquid media. They were all incubated at 296 for 96 h and observed at defined time points. The optical density was recorded to generate the growth curves in liquid media. Morphologic changes of colonies in the solid media were determined under the light microscope. Results. The clinical isolates of Fusarium solani showed stronger reproductive capacity in the abominable nutritional condition. Besides, when the glucose concentration in the medium was consistent with the glucose concentration of aqueous in diabetic patients, the clinical isolates would show the biological features of quicker growth rate and stronger reproductive capacity. Conclusions. Nitrogen source is essential for fungus reproduction. The clinical isolates showed stronger environmental adaptability under different nutritional conditions and more sensitive to environmental changes.

Download Full-text

Biosynthesis and optimization of silver nanoparticles by endophytic fungus Fusarium solani

Materials Letters ◽

10.1016/j.matlet.2014.06.143 ◽

2014 ◽

Vol 132 ◽

pp. 428-431 ◽

Cited By ~ 16

Author(s):

B. Sogra Fathima ◽

Raj Mohan Balakrishnan

Keyword(s):

Silver Nanoparticles ◽

Fusarium Solani ◽

Endophytic Fungus

Download Full-text

14,15-Epoxyeicosatrienoic acid inhibits prostaglandin E2 production in vascular smooth muscle cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1998.275.6.h2113 ◽

1998 ◽

Vol 275 (6) ◽

pp. H2113-H2121 ◽

Cited By ~ 14

Author(s):

Xiang Fang ◽

Steven A. Moore ◽

Lynn L. Stoll ◽

Gretchen Rich ◽

Terry L. Kaduce ◽

...

Keyword(s):

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cells ◽

Vascular Smooth Muscle Cells ◽

Competitive Inhibition ◽

Muscle Cells ◽

Epoxyeicosatrienoic Acid ◽

Growth Responses ◽

Enhanced Production ◽

Cytochrome P 450

14,15-Epoxyeicosatrienoic acid (EET), a cytochrome P-450 epoxygenase product of arachidonic acid (AA), reduced PGE2 formation by 40–75% in porcine aortic and murine brain microvascular smooth muscle cells. The inhibition was reversed 6–10 h after removal of 14,15-EET from the medium and was regioisomeric specific; 8,9-EET produced a smaller effect, whereas 11,12- and 5,6-EET were ineffective. Although the cells converted 14,15-EET to 14,15-dihydroxyeicosatrienoic acid (14,15-DHET), 14,15-DHET did not inhibit PGE2formation, and the 14,15-EET-induced inhibition was potentiated by 4-phenylchalcone oxide, an epoxide hydrolase inhibitor. The inhibition occurred when substrate amounts of AA were used and was not accompanied by enhanced production of other PGs, suggesting an effect on PGH synthase; however, in murine cells, 14,15-EET did not reduce PGH synthase mRNA or protein. Moreover, the 14,15-EET-induced decrease in PGE2 production was overcome by increasing the concentration of AA, but not oleic acid (which is not a substrate for PGH synthase). These findings suggest that 14,15-EET competitively inhibits PGH synthase activity in vascular smooth muscle cells. The 14,15-EET-induced inhibition of PGE2 production resulted in potentiation of platelet-derived growth factor-induced smooth muscle cell proliferation, suggesting that the competitive inhibition of PGH synthase by 14,15-EET can affect growth responses in smooth muscle cells.

Download Full-text

Enhanced production of unusual triterpenoids from Kadsura angustifolia fermented by a symbiont endophytic fungus, Penicillium sp. SWUKD4.1850

Phytochemistry ◽

10.1016/j.phytochem.2018.11.005 ◽

2019 ◽

Vol 158 ◽

pp. 56-66 ◽

Cited By ~ 7

Author(s):

Dan Qin ◽

Weiyun Shen ◽

Junqi Wang ◽

Meijun Han ◽

Fangni Chai ◽

...

Keyword(s):

Endophytic Fungus ◽

Penicillium Sp ◽

Enhanced Production

Download Full-text

Reduction of soil-borne pathogen Fusarium solani reproduction in soil enriched with phenolic acids by inoculation of endophytic fungus Phomopsis liquidambari

BioControl ◽

10.1007/s10526-016-9773-9 ◽

2016 ◽

Vol 62 (1) ◽

pp. 111-123 ◽

Cited By ~ 7

Author(s):

Xing-Guang Xie ◽

Chuan-Chao Dai ◽

Xiao-Gang Li ◽

Jia-Rui Wu ◽

Qing-Qian Wu ◽

...

Keyword(s):

Phenolic Acids ◽

Fusarium Solani ◽

Endophytic Fungus

Download Full-text

Antimicrobial Additives from Endophytic Fungus Fusarium solani of Ficus carica

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.178-181.783 ◽

2012 ◽

Vol 178-181 ◽

pp. 783-786 ◽

Cited By ~ 2

Author(s):

Hong Chi Zhang ◽

Yang Min Ma ◽

Rui Liu

Keyword(s):

Fusarium Solani ◽

Endophytic Fungus ◽

Spectroscopic Analysis ◽

Screening Program ◽

Ficus Carica ◽

Antifungal Activities ◽

Antibacterial And Antifungal Activities ◽

Antibacterial And Antifungal

Within our screening program for antimicrobial additives, six compounds were isolated from an endophytic fungus Fusarium solani of Ficus carica. Their structures were elucidated on the basis of spectroscopic analysis. Moreover, all of them showed potent antibacterial and antifungal activities.

Download Full-text