Detection of vinblastine and related bioactive compounds from culture extracts of endophytic fungi of Catharanthus roseus

• Published in: World journal of microbiology & biotechnology 2024-09, Vol.40 (9), p.278, Article 278
• Main Authors: Khan, Uzma, Gond, Surendra Kumar
• Format: Article
• Language: English
• Subjects: Applied Microbiology; Aspergillus; Bioactive compounds; Biochemical tests; Biochemistry; Biomedical and Life Sciences; Biosynthetic Pathways; Biotechnology; Catharanthus - microbiology; Chromatography; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Curvularia lunata; Endophytes; Endophytes - isolation & purification; Endophytes - metabolism; Environmental Engineering/Biotechnology; Fungi; Fungi - classification; Fungi - genetics; Fungi - isolation & purification; Fungi - metabolism; High performance liquid chromatography; Life Sciences; Liquid chromatography; Mass Spectrometry; Mass spectroscopy; Microbiology; Molecular ions; Plant Leaves - microbiology; Retention time; SDG3 Good Health and Wellbeing; Sequence analysis; Thin layer chromatography; Vinblastine; Vinblastine - metabolism
• ISSN: 0959-3993; 1573-0972; 1573-0972
• DOI: 10.1007/s11274-024-04078-9

This study investigates the synthesis of vinblastine by endophytic fungi isolated from leaf of C. roseus . A total of 10 endophytic fungi were selected for secretion of vinca alkaloids based on the initial screening by biochemical tests and thin-layer chromatography (TLC). Out of these ten, only four fungal extracts showed positive results for presence of vinblastine at same retention time (10 min.) compared to reference compound on HPLC analysis. The detected concentration of vinblastine was maximum (17 µg/ml) in isolate no. CRL 22 followed by CRL 52, CRL 17 and CRL 28. To validate the presence of vinblastine, ultra-high-performance liquid chromatography coupled with high-resolution accurate mass spectrometry (HRMS) was employed. This analysis confirmed the presence of anhydrovinblastine, a precursor of vinblastine through the detection of molecular ions at m/z 793.4185 in extract of CRL 17. In addition to anhydrovinblastine, the intermediate compounds essential to the biosynthetic pathway of vinblastine were also detected in the extract of CRL 17. These host-origin compounds strongly suggest the presence of a biosynthetic pathway within the endophytic fungus. Based on morphological observation and sequence analysis of the ITS region of rDNA, endophytic fungi were identified as Alternaria alternata (CRL 17), Curvularia lunata (CRL 28), Aspergillus terrus (CRL 52), and Aspergillus clavatonanicus (CRL 22).