Metabolomics and dereplication studies of entophytic metabolites from some Egyptian medicinal plants in the search for new potential anti-cancer and antimicrobial drugs

Ahmed Fares Said Tawfike

Research output: ThesisDoctoral Thesis

Abstract

Endophytes are microbes that inhabit living, internal tissues of plants without causing any immediate, apparent negative effects. Endophytic fungi associated with medicinal plants represent a potential source of novel chemistry and biology. This study involved isolation of three endophytic fungal strains from two Egyptian medicinal plants, Terminalia laxiflora and Markhamia platicalyx. Identification of the strains has been achieved through molecular biological methods. Metabolomic profiling, using 2D-NMR and HR-ESIFTMS were done at different stages of the growth phase for both solid and liquid culture media. Dereplication studies were accomplished by utilizing the MZmine 2.10 software with aid of the AntiBase and DNP databases. The optimized method in terms of media, incubation time, and maximum production of bioactive compounds were taken into account for the scale-up. The chemometric analysis using SIMCA-P v13 software was accomplished to compare the active and inactive extracts and/or fractions in order to highlight the bioactive metabolites at earlier stages. The bioassay-guided fractionation was established using flash chromatography that employed BUCHI®, Biotage® and Reveleris® flash systems. The cytotoxicity of extracts, fractions and the isolated compounds was tested against both chronic myelogenous leukemia (K562) and prostate cancer (PC3) cell lines. The antimicrobial activity was investigated against Trypanosoma brucei and Mycobacterium marinum ATCCBAA 535. This work led to isolation of three new natural compounds and 34 known endophytic metabolites from Aspergillus aculeatus, A. flocculus and Curvularia sp. The isolated fungal compounds were elucidated using 1D and 2D-NMR experiments and HR-ESIFTMS with aid of DNP databases. Secalonic acids D, B, F and C from A. aculeatus; phomaligol A, p-hydroxybenzaldehyde, 4-hydroxyphenyl-methoxy-acetic acid, mellein derivatives like mellein, trans-4-hydroxymellein and 5-hydroxymellien and diorcinol from A. flocculus; dipeptide (N-acetyl-phenylalanine-L-phenylalanine) from Curvularia sp., were the most significant bioactive metabolites. Metabolomics has been shown to be a powerful facilitator in the discovery of natural products, which are considered an excellent source for novel leads, and even more, as a means to highlight novel targets.
LanguageEnglish
Awarding Institution
  • Strathclyde Institute Of Pharmacy And Biomedical Sciences
Supervisors/Advisors
  • Edrada-Ebel, Ruangelie, Supervisor
  • Watson, David, Supervisor
Award date23 Feb 2015
Place of PublicationGlasgow
Publisher
Publication statusPublished - 23 Feb 2015

Fingerprint

Metabolomics
Medicinal Plants
Phenylalanine
Software
Mycobacterium marinum
Terminalia
Databases
Endophytes
Trypanosoma brucei brucei
Dipeptides
Aspergillus
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Biological Products
Acetic Acid
Biological Assay
Pharmaceutical Preparations
Culture Media
Chromatography
Neoplasms
Prostatic Neoplasms

Keywords

  • fungal endophyte
  • metabolomics
  • dereplication
  • NMR
  • MS
  • Egyptian medicinal plants

Cite this

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title = "Metabolomics and dereplication studies of entophytic metabolites from some Egyptian medicinal plants in the search for new potential anti-cancer and antimicrobial drugs",
abstract = "Endophytes are microbes that inhabit living, internal tissues of plants without causing any immediate, apparent negative effects. Endophytic fungi associated with medicinal plants represent a potential source of novel chemistry and biology. This study involved isolation of three endophytic fungal strains from two Egyptian medicinal plants, Terminalia laxiflora and Markhamia platicalyx. Identification of the strains has been achieved through molecular biological methods. Metabolomic profiling, using 2D-NMR and HR-ESIFTMS were done at different stages of the growth phase for both solid and liquid culture media. Dereplication studies were accomplished by utilizing the MZmine 2.10 software with aid of the AntiBase and DNP databases. The optimized method in terms of media, incubation time, and maximum production of bioactive compounds were taken into account for the scale-up. The chemometric analysis using SIMCA-P v13 software was accomplished to compare the active and inactive extracts and/or fractions in order to highlight the bioactive metabolites at earlier stages. The bioassay-guided fractionation was established using flash chromatography that employed BUCHI{\circledR}, Biotage{\circledR} and Reveleris{\circledR} flash systems. The cytotoxicity of extracts, fractions and the isolated compounds was tested against both chronic myelogenous leukemia (K562) and prostate cancer (PC3) cell lines. The antimicrobial activity was investigated against Trypanosoma brucei and Mycobacterium marinum ATCCBAA 535. This work led to isolation of three new natural compounds and 34 known endophytic metabolites from Aspergillus aculeatus, A. flocculus and Curvularia sp. The isolated fungal compounds were elucidated using 1D and 2D-NMR experiments and HR-ESIFTMS with aid of DNP databases. Secalonic acids D, B, F and C from A. aculeatus; phomaligol A, p-hydroxybenzaldehyde, 4-hydroxyphenyl-methoxy-acetic acid, mellein derivatives like mellein, trans-4-hydroxymellein and 5-hydroxymellien and diorcinol from A. flocculus; dipeptide (N-acetyl-phenylalanine-L-phenylalanine) from Curvularia sp., were the most significant bioactive metabolites. Metabolomics has been shown to be a powerful facilitator in the discovery of natural products, which are considered an excellent source for novel leads, and even more, as a means to highlight novel targets.",
keywords = "fungal endophyte, metabolomics, dereplication, NMR, MS, Egyptian medicinal plants",
author = "Tawfike, {Ahmed Fares Said}",
year = "2015",
month = "2",
day = "23",
language = "English",
publisher = "University of Strathclyde",
school = "Strathclyde Institute Of Pharmacy And Biomedical Sciences",

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Metabolomics and dereplication studies of entophytic metabolites from some Egyptian medicinal plants in the search for new potential anti-cancer and antimicrobial drugs. / Tawfike, Ahmed Fares Said.

Glasgow : University of Strathclyde, 2015.

Research output: ThesisDoctoral Thesis

TY - THES

T1 - Metabolomics and dereplication studies of entophytic metabolites from some Egyptian medicinal plants in the search for new potential anti-cancer and antimicrobial drugs

AU - Tawfike, Ahmed Fares Said

PY - 2015/2/23

Y1 - 2015/2/23

N2 - Endophytes are microbes that inhabit living, internal tissues of plants without causing any immediate, apparent negative effects. Endophytic fungi associated with medicinal plants represent a potential source of novel chemistry and biology. This study involved isolation of three endophytic fungal strains from two Egyptian medicinal plants, Terminalia laxiflora and Markhamia platicalyx. Identification of the strains has been achieved through molecular biological methods. Metabolomic profiling, using 2D-NMR and HR-ESIFTMS were done at different stages of the growth phase for both solid and liquid culture media. Dereplication studies were accomplished by utilizing the MZmine 2.10 software with aid of the AntiBase and DNP databases. The optimized method in terms of media, incubation time, and maximum production of bioactive compounds were taken into account for the scale-up. The chemometric analysis using SIMCA-P v13 software was accomplished to compare the active and inactive extracts and/or fractions in order to highlight the bioactive metabolites at earlier stages. The bioassay-guided fractionation was established using flash chromatography that employed BUCHI®, Biotage® and Reveleris® flash systems. The cytotoxicity of extracts, fractions and the isolated compounds was tested against both chronic myelogenous leukemia (K562) and prostate cancer (PC3) cell lines. The antimicrobial activity was investigated against Trypanosoma brucei and Mycobacterium marinum ATCCBAA 535. This work led to isolation of three new natural compounds and 34 known endophytic metabolites from Aspergillus aculeatus, A. flocculus and Curvularia sp. The isolated fungal compounds were elucidated using 1D and 2D-NMR experiments and HR-ESIFTMS with aid of DNP databases. Secalonic acids D, B, F and C from A. aculeatus; phomaligol A, p-hydroxybenzaldehyde, 4-hydroxyphenyl-methoxy-acetic acid, mellein derivatives like mellein, trans-4-hydroxymellein and 5-hydroxymellien and diorcinol from A. flocculus; dipeptide (N-acetyl-phenylalanine-L-phenylalanine) from Curvularia sp., were the most significant bioactive metabolites. Metabolomics has been shown to be a powerful facilitator in the discovery of natural products, which are considered an excellent source for novel leads, and even more, as a means to highlight novel targets.

AB - Endophytes are microbes that inhabit living, internal tissues of plants without causing any immediate, apparent negative effects. Endophytic fungi associated with medicinal plants represent a potential source of novel chemistry and biology. This study involved isolation of three endophytic fungal strains from two Egyptian medicinal plants, Terminalia laxiflora and Markhamia platicalyx. Identification of the strains has been achieved through molecular biological methods. Metabolomic profiling, using 2D-NMR and HR-ESIFTMS were done at different stages of the growth phase for both solid and liquid culture media. Dereplication studies were accomplished by utilizing the MZmine 2.10 software with aid of the AntiBase and DNP databases. The optimized method in terms of media, incubation time, and maximum production of bioactive compounds were taken into account for the scale-up. The chemometric analysis using SIMCA-P v13 software was accomplished to compare the active and inactive extracts and/or fractions in order to highlight the bioactive metabolites at earlier stages. The bioassay-guided fractionation was established using flash chromatography that employed BUCHI®, Biotage® and Reveleris® flash systems. The cytotoxicity of extracts, fractions and the isolated compounds was tested against both chronic myelogenous leukemia (K562) and prostate cancer (PC3) cell lines. The antimicrobial activity was investigated against Trypanosoma brucei and Mycobacterium marinum ATCCBAA 535. This work led to isolation of three new natural compounds and 34 known endophytic metabolites from Aspergillus aculeatus, A. flocculus and Curvularia sp. The isolated fungal compounds were elucidated using 1D and 2D-NMR experiments and HR-ESIFTMS with aid of DNP databases. Secalonic acids D, B, F and C from A. aculeatus; phomaligol A, p-hydroxybenzaldehyde, 4-hydroxyphenyl-methoxy-acetic acid, mellein derivatives like mellein, trans-4-hydroxymellein and 5-hydroxymellien and diorcinol from A. flocculus; dipeptide (N-acetyl-phenylalanine-L-phenylalanine) from Curvularia sp., were the most significant bioactive metabolites. Metabolomics has been shown to be a powerful facilitator in the discovery of natural products, which are considered an excellent source for novel leads, and even more, as a means to highlight novel targets.

KW - fungal endophyte

KW - metabolomics

KW - dereplication

KW - NMR

KW - MS

KW - Egyptian medicinal plants

M3 - Doctoral Thesis

PB - University of Strathclyde

CY - Glasgow

ER -