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Title: Diversity of taxol producing endophytic fungi from taxus baccata and process optimization for taxol production
Researcher: Garyali, Sanjog
Guide(s): Reddy, M. Sudhakara and Kumar, Anil
Keywords: Agrobacterium tumefaciens
Endophytic fungi
Engineering and Technology
Taxus baccata
University: Thapar Institute of Engineering and Technology
Completed Date: 2014
Abstract: Endophytic fungi represent an under explored resource of novel lead compounds and have the capacity to produce diverse classes of plant secondary metabolites. In the present study, we investigated the diversity of taxol-producing endophytic fungi from Taxus baccata L. subsp. wallichiana (Zucc.) Pilger. A total of 60 fungal endophytes were isolated from the inner bark (phloem-cambium)of T. baccata,collected from different locations of the northern Himalayan region of India. Two key genes, DBAT (10-deacetylbaccatin III-10-O-acetyl transferase) and BAPT (C-13 phenylpropanoid side chain-CoA acyltransferase), involved in taxol biosynthesis were used as molecular markers for screening of the taxol producing strains. Five different endophytic species gave positive amplification hits by molecular marker screening with the bapt gene. These fungi were characterized based on morphological characters and internal transcribed spacer (ITS) sequence analysis and identified as: Fusarium redolens (TBPJ-B), Gibberella avenacea (C-1), Paraconiothyrium brasiliense (TBPJ-13), Microdiplodia sp. (TBPJ-A) and Fusarium tricinctum (B-7), The taxol-producing capability of these endophytic fungi was validated by HPLC-MS. The highest yield of taxol was found to be 66.25 and#956;g/L by Fusarium redolens compared to other four strains. The amount of taxol produced by other four strains in S-7 liquid medium are TBPJ-A (27.40 µg/L), B-7 (23.47 µg/L), TBPJ-13 (19.60 µg/L) and C-1 (11.03 µg/L). The antitumour activity of the fungal taxol was tested by potato disc tumor induction assay using Agrobacterium tumefaciens as the tumor induction agent. This assay depicted that fungal taxol from all the endophytes successfully inhibited tumor formation in potato discs just like authentic taxol while it did not affect bacterial viability in any case. These results confirmed that all fungal extracts tested in this study had similar antineoplastic activity as that of paclitaxel.
Pagination: xix, 243p.
Appears in Departments:Department of Biotechnology

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