The most promising anti-tumor agent developed in the past three decades is Taxol. It is proven to be effective against many cancers. It is necessary to isolate pharmacologically potent endophytic microbial strains from medicinal plants with special reference to Taxol production. In the current study, endophytic fungi were isolated from the bark of the medicinal plant, Salacia oblonga. The isolated endophytes were identified morphologically, and further characterized by ITS-PCR using genomic DNA samples, later the products were sequenced for identification and phylogenetic linkage mapping. The samples were screened for the potential to produce Taxol or taxanes, employing PCR. The resulted data have been sequenced to confirm the presence of the two genes implicated in Taxol biosynthesis, 10-deacetylbaccatin III-10-O-acetyl transferase (DBAT) and C-13 phenylpropanoid side chain-CoA acyltransferase (BAPT). Seven samples showed the amplicons of DBAT gene and one showed the amplicons of BAPT gene. Sequencing of these products was carried out, of which one sample has revealed sequence homology to the original DBAT gene from Taxus. The present work confirms and substantiates the potential of genomic mining approach to discover novel Taxol-producing endophytic fungi.

No abstract

Key indicators: single-crystal X-ray study; T = 203 K; mean (C-C) = 0.003 Å; R factor = 0.031; wR factor = 0.081; data-to-parameter ratio = 26.2.

USP official UV-VIS spectrophotometric method for the evaluation of antioxidant activity by scavenging the DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical was not suitable to determine the antioxidant activity of Berry fruit juice freeze dried extract due to interferences from the matrix. Interference was also observed from the color pigments of the Berry fruit juice freeze dried extract. This has leads to the development of a specific analytical method for evaluation of the DPPH free radical scavenging activity of Berry fruit juice freeze dried extract. RP-HPLC method was developed on a Phenomenex Luna-C18 Column (250 mm × 4 mm, 5 µM) with mobile phase methanol and water mixed online in the ratio of 80:20 (v/v), pumped at a flow rate of 1 mL/min. DPPH peaks were quantified at wavelength 517 nm. Method was standardized using the anti-oxidant ascorbic acid. The 50% free radical scavenging activity (IC 50 ) determined by the novel HPLC method was correlating with the results obtained by USP UV-VIS spectrophotometric method. While the USP UV-VIS spectrophotometric method failed to estimate the free radical scavenging activity of Berry fruit juice freeze dried extract, whereas the developed HPLC method was specific, sensitive, robust, accurate and precise and rapid in the determination.The objective of the present study is to develop, standardize and compare the DPPH-HPLC method with that of spectrophotometric method using known antioxidants, and to evaluate its application to determine the antioxidant activity of selected berry fruit juice freeze dry powder as a means of rapid screening methodology.

Context: Endophytes colonizing medicinal plants are diverse, constituting a rich bioresource for novel natural products. Objective: Myrothecium sp. isolate M1-CA-102 was the most promising among the 16 Myrothecium isolates screened. The bioactive potential of the crude extract from the Calophyllum apetalum Willd. endophytic Myrothecium sp. (Alb. & Schwein.) Ditmar (Incertae sedis) isolate M1-CA-102 and its thin layer chromatography (TLC) fractions were screened based on antioxidant, anti-inflammatory, antimicrobial activities, and cytotoxicity. Materials and methods: The antioxidant activity was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2 0 -azinobis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging capacities. Further, 15-lipoxygenase (15-LOX) and human cyclooxygenase-2 (COX-2) inhibition were assessed at different concentrations (25, 50, and 100 mg/mL for the crude extract, 5, 25, and 50 mg/mL for the TLC fractions). DNA-nicking assay as an indicator of the capacity of extracts to scavenge hydroxyl radical was recorded at a concentration of 50 mg/mL. Cell cytotoxicity was recorded by colorimetric 3-(4,5-dimethylthylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Antibacterial (Bacillus subtilis) and antiCandida (Candida albicans) assays were performed by the microdilution method. Results: The DPPH and ABTS IC 50 values of M1-CA-102 extract were 10 and 6 mg/mL compared with 6.1 and 7.03 mg/mL for the positive control quercetin. The cytotoxicity IC 50 value of M1-CA-102 extract was 37 mg/mL, while the M-I TLC fraction was 21 mg/mL. The M1-CA-102 extract gave an IC 50 value of 58 and 8 mg/mL for 15-LOX and COX-2, respectively. The MIC values for antimicrobial activity for M1-CA-102 extract ranged from 35 to 54 mg/mL, while for the TLC fractions, it ranged from 91 to 515 mg/mL. Conclusion:The results indicate that Myrothecium M1-CA-102 isolated from C. apetalum is a potential source of natural metabolites of pharmaceutical importance.