Production of Taxol by Endophytic Fungi Isolated from Roots of Himalayan Yew (Taxus wallichiana Zucc.)


  • Priyanka Adhikari G.B. Pant National Institute of Himalayan Environment, Kosi-Katarmal, Almora-263643, Uttarakhand, India
  • Mithilesh Singh G.B. Pant National Institute of Himalayan Environment, Kosi-Katarmal, Almora-263643, Uttarakhand, India
  • Anita Pandey G.B. Pant National Institute of Himalayan Environment, Kosi-Katarmal, Almora-263643, Uttarakhand, India



Taxus wallichiana, endophytes, taxol, Aspergillus, Penicillium.


Taxol® (generic name – Paclitaxel), the most promising chemotherapeutic
agent was isolated from bark of different Taxus sp. As Taxus species are
threatened with extinction (endangered in Himalaya), thus it is imperative
to develop alternate and sustainable method for commercialization and scale
up production of paclitaxel. In this respect, physical and chemical parameters
are effective and important key points for active compound production par-
ticularly by using endophytic microbes. In the present study, five endophytic
fungi isolated from the roots of Taxus wallichiana, are tested for paclitaxel
production using biochemical and molecular methods. Subsequently, effect
of physico-chemical parameters like temperature, pH, incubation time, and
medium constituents i.e., salt concentration, carbon and nitrogen sources on
paclitaxel production were also analyzed. Among isolates, two of the fungi
viz. GBPI TWR F1 (Penicillium sp.) and GBPI TWR F5 (Aspergillus sp.)
were found to be paclitaxel producing. The genomic DNA samples were sequenced to confirm the presence of two genes viz. 10-deacetylbaccatin
III-10-O-acetyl transferase (DBAT) and C-13 phenylpropanoid side chain-
CoA acyltransferase (BAPT), implicated in paclitaxel biosynthesis. Both
the endophytes showed the amplicons of DBAT and BAPT genes. Results
revealed that after optimization of medium components and physical condi-
tion, paclitaxel production was increased in both the endophytes, maximum
paclitaxel production i.e., 5.45 ± 0.98 mg/L was obtained by GBPI TWR F5
(Aspergillus sp.) following 10 days of incubation at 15◦C in optimized S7
liquid medium composition


Download data is not yet available.

Author Biographies

Priyanka Adhikari, G.B. Pant National Institute of Himalayan Environment, Kosi-Katarmal, Almora-263643, Uttarakhand, India

Priyanka Adhikari is Research Associate at National Institute of Pharma-
ceutical Education and Research (NIPER), Guwahati-Assam, India. Before
Joining NIPER, Dr. Adhikari worked as Junior and Senior Project Fellow
in National Mission on Himalayan Studies (NMHS) Program of Min-
istry of Environment, Forest and Climate Change, Govt. of India at G.B.
Pant National Institute of Himalayan Environment, Almora – Uttarakhand,
India. Dr. Adhikari has M.Sc. in Microbiology and Ph.D. in Biotechnology
from Kumaun University, Nainital-Uttarakhand, India. Her research interests
include plant-microbe interactions and characterization, and identification
of microbe and plant based bioactive compounds and drug formulation. Dr
Adhikari has been recognized for National Award (twice) from MoEF &
CC Himalayan Researchers Consortium and National Medicinal Plant Board
(NMPB) Govt. of India for her work on microbiological and biochemical
aspects of Taxus wallichiana. Later, she received the Young Scientist Travel
Grant to present her work in 8th Conference “FEMS 2019” at Glasgow
Scotland, United-Kingdom. She has published more than 20 peer reviewed
papers in National and International Journals.

Mithilesh Singh, G.B. Pant National Institute of Himalayan Environment, Kosi-Katarmal, Almora-263643, Uttarakhand, India

Mithilesh Singh is working as Scientist at Centre for Environmental Assess-
ment and Climate Change, G.B. Pant National Institute of Himalayan Envi-
ronment (GBP-NIHE), Almora, Uttarakhand. She has done post graduation in
Botany from Banaras Hindu University and PhD in Biotechnology from IIT Guwahati, Assam. Her field of specialization includes plant biotechnology
and bioprospection. She has published 55 research papers, chapters in edited
books/proceedings and scientific popular articles having over 520 citation and
12 h index. Dr. Singh has received DBT BioCARe Women Scientist Award
in 2014.

Anita Pandey, G.B. Pant National Institute of Himalayan Environment, Kosi-Katarmal, Almora-263643, Uttarakhand, India

Anita Pandey is presently affiliated with the Department of Biotechnology,
Graphic Era (Deemed to be University, Dehradun, Uttarakhand, India. Previ-
ously, she was Scientist at G.B. Pant National Institute of Himalayan Environ-
ment, Almora, India. Dr. Pandey has extensively worked on Bioprospection
of Microbial Diversity of Indian Himalayan region (IHR). Her research areas
include- Extremophiles, Rhizosphere, Biodegradation, Fermented Foods, and
Her scientific contributions have been recognized at State, National, and
International levels. She has been recipient of National Bioscientist Award by
Department of Biotechnology, Govt. of India and Vishisht Mahila Vaigyanik
Sammaan (Eminent Woman Scientist Recognition) by Uttarakhand Science,
Education and Research Center, Department of Science and Technology,
Govt. of Uttarakhand, India.
Dr. Pandey has hosted CV Raman International Fellowship for African
Researchers twice (Morocco and Egypt). A new archeal species (Nantrono-
coccus pandeyae sp. nov.) has been named in her honour for her extensive
contribution to microbial diversity of IHR (Current Microbiology, vol. 79,
Article number 51, 2022).


Abdel-Fatah, S.S., El-Batal, A.I., El-Sherbiny, G.M., Khalaf, M.A.,

El-Sayed, A.S. (2021). Production, bioprocess optimization and

g-irradiation of Penicillium polonicum, as a new paclitaxel producing

endophyte from Ginkgo biloba. Biotechnology Report. 30, e00623.

Adhikari, P., Pandey, A. (2019). Phosphate solubilization potential of endo-

phytic fungi isolated from Taxus wallichiana Zucc. roots. Rhizosphere.

, 2–9.

Adhikari, P., Pandey, A. (2018). Diversity of endophytic fungi associated

with Himalayan yew (Taxus wallichiana Zucc.) roots. Proceedings of

Himalayan Researchers Consortium. 1, 165–173.

Afshari, M., Shahidi, F., Mortazavi, S.A., Tabatabai, F., Eshaghi, Z. (2015).

Investigating the influence of pH, temperature and agitation speed on

yellow pigment production by Penicillium aculeatum ATCC 10409.

Natural Products Research. 29, 1300–1306.

Andualem, B., Gessesse, A. (2013). Production of microbial medium from

defatted brebra (Milletia ferruginea) seed flour to substitute commercial

peptone agar. Asian Pacific Journal of Tropical Biomedical. 3, 790–797.

Barbuti, A.M., Chen, Z.S. (2015) Paclitaxel through the ages of anticancer

therapy: Exploring its role in chemoresistance and radiation therapy.

Cancers. 7, 2360–2371.

Choi, H.K., Kim, S.I., Son, J.S., Hong, S.S., Lee, H.S., Chung, I.S., Lee, H.J.

(2000) Intermittent maltose feeding enhances paclitaxel production in

suspension culture of Taxus chinensis cells. Biotechnology Letters. 22,


Costa, E., Teixido, N., Usall, J., Atares, E., Vinas, I. (2002). The effect of

nitrogen and carbon sources on growth of the biocontrol agent Pan-

toea agglomerans strain CPA-2. Letters in Applied Microbiology. 25,


Chen, Y.J., Zhang, Z., Wang, Y., Su, Y., Zhang, R. (2003). Screening

endophytic fungus to produce paclitaxel from Taxus yunnanensis.

Biotechnology. 13, 10–11.

El-Bialy, H.A., El-Bastawisy, H. (2020). Elicitors stimulate paclitaxel pro-

duction by endophytic fungi isolated from ecologically altered Taxus

baccata. Journal of Radiation Research and Applied Sciences. 13,


El-Sayed, A.S.A., Safan, S., Mohamed, N.Z., Shaban, L., Ali, G.S., Sitohy,

M.Z. (2018) Induction of paclitaxel biosynthesis by Aspergillus terreus,

Production of Taxol by Endophytic Fungi Isolated from Roots 211

endophyte of Podocarpus gracilior Pilger. upon intimate interaction

with the plant endogenous microbes. Process Biochemistry. 71, 31–40.

El-Sayed, R., Ahmed, A.S., Hassan, I.A., Ismail, A.A., El-Din, A.Z.A.K.,

(2020). Semi-continuous production of the anticancer drug paclitaxel

by Aspergillus fumigatus and Alternaria tenuissima immobilized in

calcium alginate beads. Bioprocess and Biosystems Engineering. 43,


Kumar, P., Singh, B., Thakur, V., Thakur, A., Thakur, N., Pandey, D., Chand,

D. (2019). Hyper production of paclitaxel from Aspergillus fumigatus,

an endophytic fungus isolated from Taxus sp. of the northern Himalayan

region. Biotechnology Reports. 24, 1–13.

Kusari, S., Singh, S., Jayabaskaran, C. (2014). Rethinking production

of paclitaxel (paclitaxel) using endophyte biotechnology. Trends in

Biotechnol. 32, 304–311.

Li, D., Fu, D.W., Zhang, Y., Ma, X., Gao, L., Wang, X., Zhau, D., Zhao, K.

(2017). Isolation, purification, and identification of paclitaxel and related

taxanes from paclitaxel-producing fungus Aspergillus niger subsp. taxi.

Journal of Microbiology and Biotechnology. 27, 1379–1385.

Nadeem, M., Rikhari, H.C., Kumar, A., Palni, L.M.S., Nandi, S.K. (2002).

Paclitaxel content in the bark of Himalayan Yew in relation to tree age

and sex. Phytochemistry. 60, 627–631.

Naik, B.S. (2018). Developments in paclitaxel production through endo-

phytic fungal biotechnology: A review. Oriental Pharmacy and Exper-

imental Medicine. 19, 1–13.

NATA, Technical note 17 (2013). Guidelines for the validation and verifica-

tion of quantitative and qualitative test methods. National Association of

Testing Authorities, Australia. pp. 17–18.

Pandey, N., Jain, R., Pandey, A., Tamta, S. (2018). Optimisation and char-

acterisation of the orange pigment produced by a cold adapted strain

of Penicillium sp. (GBPI P155) isolated from mountain ecosystem.

Mycology. 9, 81–92.

Poupat, C., Hook, I., Gueritte, F., Ahond, A., Guenard, D., Adeline, M.T.,

Wang, X.P., Dempsey, D., Breuillet, S., Potier, P. (2000). Neutral and

basic taxoid contents in the needles of Taxus species. Planta Medica. 66,


Qiao, W., Ling, F., Yu, L., Huang, Y., Wang, T. (2017). Enhancing pacli-

taxel production in a novel endophytic fungus, Aspergillus aculeatus

Tax-6, isolated from Taxus chinensis var. mairei. Fungal Biology. 121,


P. Adhikari et al.

Qiao, W., Tang, T., Ling, F. (2021). Comparative transcriptome analysis of a

paclitaxel-producing endophytic fungus, Aspergillus aculeatinus Tax-6,

and its mutant strain. Scientific Reports. 10, 10558.


Roopa, G., Madhusudhan. M.C., Sunil, K.C.R., Lisa, N., Calvin, R., Poorn-

ima, R., Zeinab, N., Kini, K.R., Prakash, H.S., Geetha, N. (2015).

Identification of paclitaxel producing endophytic fungi isolated from

Salacia oblonga through genomic mining approach. Journal of Genetic

Engineering and Biotechnology. 13, 119–127.

Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A., Allard, R.W. (1984).

Ribosomal DNA spacer-length polymorphism in barley: Mendelian

inheritance, chromosomal location and population dynamics. Proceed-

ings of National Academy of Science USA. 81, 8014–8018.

Sanchez, S., Chavez, A., Ferero, A., Huante, Y.G., Romero, A., Sanchez,

M., Richa, D., Sanchez, B., Avalos, M., Trampe, S.G., Sanoja, R.R.,

Langley, E., Ruiz, B. (2010). Carbon source regulation of antibiotic

production. Journal of Antibiotics. 63, 442–459.

Sengul, U. (2016). Comparing determination methods of detection and quan-

tification limit for oflatoxin analysis in hazulnut. Journal of Food Drug

and Analysis. 24, 56–62.

Sharma, B., Jha, D.K. (2015). Role of nitrogen sources in regulation of

fungal secondary metabolism. In: Gupta VK, Mach RL, Sreenivas-

aprasad S (eds) Fungal biomolecules: Sources, applications and recent

developments, 1st edn. John Wiley & Sons, Ltd., pp. 213–224.

Shreshta, K., Strobel, G.A., Shrivastava, S.P., Gewali, M.B. (2001). Evidence

for paclitaxel from three new endophytic fungi of Himalayan Yew of

Nepal. Planta Medica. 67, 374–376.

Shu, C.H. (2007). Fungal fermentation for medicinal plants. In: Yang ST (ed)

Bioprocessing for value-added products from renewable resources, 1st

edn. Elsevier, pp. 447–463.

Somjaipeng, S., Median, A., Kwasna, H., Ordaz-Ortiz, J., Magan, N. (2015).

Isolation, identification and ecology of growth and paclitaxel pro-

duction by an endophytoc strain of Paraconiothyrium variabile from

English Yew tree (Taxus baccata). Brazilian Mycological Society. 119,


Somjaipeng, S., Medina, A., Magan, N. (2016). Environmental stress and

elicitors enhance paclitaxel production by endophytic strains of Para-

coniothyrium variabile and Epicoccum nigrum. Enzyme and Microbial

Technology. 90, 69–75.

Production of Taxol by Endophytic Fungi Isolated from Roots 213

Sun, D., Ran, X., Wang, J. (2008). Isolation and identification of a paclitaxel-

producing endophytic fungus from Podocarpus. Acta Microbiologica

Sinica. 48, 589–595.

Thomas, P., Farjon, A. (2011). Taxus wallichiana. The IUCN Red List of

Threatened 2011:e.T46171879A9730085.

Stierle, A., Strobel, G., Stierle, D., 1993. Taxol and taxane production by

Taxomyces andreanae, an endophytic fungus of Pacific yew. Sci. 260,


Strobel, G.A., Yang, X., Sears, J., Kramer, R., Sidhu, R., Hess, W.M. (1996).

Paclitaxel from Pestalotiopsis microspora, an endophytic fungus of

Taxus wallichiana. Microbiology. 142, 435–440.

Wani, M.C., Taylor, H.L., Wall, M.E., Coggon, O., McPhail, A.T. (1971).

Plant antitumor agents VI. The isolation and structure of paclitaxel, a

novel antileukemic and antitumor agent from Taxus brevifolia. Journal

of American Chemical Society. 92, 2325–2327.

Walker, K.D., Croteau, R.B. (2000). Molecular cloning of a 10-

deacetylbaccatin III-10-O-acetyl transferase cDNA from Taxus and

functional expression in Escherichia coli. Proceedings of National

Academic Science. 97, 583–587.

Yang, Y., Zhao, H., Barrero, R.A., Zhang, B., Sun, G., Wilson, L.W., Xie,

F., Walker, K.D., Parks, J.W., Bruce, R., Guo, G., Chen, L., Zhang, Y.,

Huang, X., Tang, Q., Liu, H., Bellgard, M.I., Qiu, D., Lai ,J., Hoffman,

A. (2014). Genome sequencing and analysis of the paclitaxel-producing

endophytic fungus Penicillium aurantiogriseum NRRL 62431. BMC

Genomics. 69.∼10.1186/1471-2164-15-69

Zaiyou, J., Li, M., Xiqiao, H. (2017). An endophytic fungus efficiently pro-

ducing paclitaxel isolated from Taxus wallichiana var. mairei. Medicine.

, e7406.




How to Cite

Adhikari, P., Singh, M., & Pandey, A. (2022). Production of Taxol by Endophytic Fungi Isolated from Roots of Himalayan Yew (Taxus wallichiana Zucc.). Journal of Graphic Era University, 10(2), 195–216.