Metabolomics and how it can be used as a tool in the identification and

production of new antibiotic from natural products

1,2 1 1
Weqas Alotaibi , Carol Clements , Rothwelle Tate , and RuAngelie Edrada-Ebel 1*

1 SIBPS, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0NR, Scotland.

* ruangelie.edrada-ebel@strath.ac.uk
2 General Administration of forensic evidences, Ministry of interior, Riyadh, Kingdom of Saudi Arabia

INTRODUCTION & AIMS

Drug discovery has been an aim of mankind since ancient times. The plant kingdom offers a renewable and unique resource for potential new drugs
discovery. Although reviewing the current literature for the importance of the plants Nerium oleander (Family:Apocynaceae) growing worldwide has
revealed many biological interests. But, as far as I know, nothing was reported concerning the endophytic fungi and their secondary metabolites as a
source of active antimicrobial natural products from the species growing in Saudi Arabia. The main goal of this study is to identify antimicrobial compounds
from endophytic secondary metabolites. Although, bioactive compounds are going to be isolated, purified and structurally elucidated using 2D-NMR and
HRESI-FTMS. These isolated compounds are going to be evaluated for their antimicrobial activities thereafter. Moreover, dereplication study and
metabolomics were done using the software MZmine coupled with DNP. This study resulted in isolation of six endophytic fungi from three different Saudi
Arabian medicinal plants. Molecular biology aspects have been used to identify the isolated endophytes. Then, cultivation of these strains was done in
both rice culture and broth culture. At different time stages (7, 15, and 30 days), I have done metabolomic profiling. Then, the most suitable method is
going to be chosen for upscaling. The upscaled medium was extracted by ethyl acetate and yielded a biological active crude extract. Crude extract was
then fractionated by different chromatographic techniques. The pure compounds were then identified using 2D-NMR and MS.

* Dereplication study and biological activity

METHODOLOGY
Alternaria alternata Antimicrobial activity of different culture
Nerium oleander Two fungal strains
extracts of Alternaria alternata
Reinoculation
Inoculation
Identification by
DNA technique

Medium up-scale

Dereplication study and

biological activity guidance
Some isolated compounds from Alternaria alternata:

Large up-scale Rice culture Liquid culture Molecular Comments

Name Structure M.Wt
formula
Incubation for 7, 15, and 30 days,
New from
at room temperature
endophytic fungi &
TLC Monitoring Turreanone 232 C9H12O7
new from the family

Apocynaceae

(22)-24a-methyl-ergost-

6,22(23)-diene-5a-8a- 428 C28H44O3 Known

RESULTS epidioxy-3B-ol

24-methylcholesta-5,22-
PCR product DNA sequencing 414 C28H46O2
Known
dien-3B,7B-diol

4-Epi-pyrenolide D 212 C10H12O5 New from the plant

species
Identification of the fungus
based on DNA sequence
Closest match Percentage cyclo-L-Phe-L-Phe 294 C18H18N2O2
Isolate Source
in GenBank identity (%) Known

airborne fungi Alternaria

Alternaria Destruxin B 649 C34H59N5O7 Known
in different caves Mogao alternata 99
alternata
Grotoes,Dunhuang, China

Conclusion
Endophytic fungi are a good source of secondary metabolites. They produce same or similar compounds as the medicinal plants do. Metabolomics
and dereplication study helped to choose the optimum and suitable method. In this study, Crude extracts of the endophyte showed selective and
significant antimicrobial activity. Isolation work afforded the compound Turreanone which is reported here for the first time from endophytic fungi as well
as from the family Apocynaceae. Although, 4-Epi-pyrenolide D is reported here for the first time from Nerium oleander species.