25
Introduction into the work with endophytic fungi as a source of potential new natural products Dipl.-Phar m. Ca talina Pérez Hemphill PhD student, Pharmacist Institut e of Phar maceuti cal Biology and Biot echnol ogy, Prof. Dr. Peter Proksch

Catalina _ Workshop1

  • Upload
    minhhoi

  • View
    225

  • Download
    0

Embed Size (px)

Citation preview

Page 1: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 1/25

Introduction into the work

endophytic fungi as a soupotential new natural pro

Dipl.-Pharm. Catalina Pérez HemphillPhD student, PharmacistInstitute of Pharmaceutical Biology and Biotechnology, Prof. Dr. Peter Proksch

Page 2: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 2/25

Overview

• Introduction

• General methods for isolation of derived fungi

• Workup of fractions

• Troubleshooting

Page 3: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 3/25

Motivation

• Discovery of Penicillin1928, by A.Flemming

Modern era of

antibiotic discovery• Fungi were now seen

as potential medicinefactories

Fig.1: Alexander

Page 4: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 4/25

Endophytic fungi

• Reside in the internal tissue ofplants

• Secondary metabolites utile

for – Plants

• second messengers

• protection against herbivores

 – Humans• Paclitaxel

Fig

Page 5: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 5/25

Marine endophytes

• Extreme habitat

adaptaion necessary

• High chemical diversity promising potential as

sources of new andbioactive natural products

Fig.3: Mang

Page 6: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 6/25

Natural product resea

• Main methods:

cultivation, extraction,isolation and structure

elucidation

• New natural productsby promoting

secondary metabolite

biosynthesis through

 – external cues

 – co-cultivation Fig.4: Cultivation and extraction

Page 7: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 7/25

Sampling plant materi

• Transport collectedmaterial in suitablecontainers

• Isolation of fungi fromplant material within1-2 days, as long asstored at 5-10°C

Fig.5: Avicennia marina

Page 8: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 8/25

Isolation of endophyt

Page 9: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 9/25

Fermentation

Page 10: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 10/25

Extraction from rice med

Page 11: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 11/25

Isolation and structure elucof bioactive secondary meta

Page 12: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 12/25

Main literature

Page 13: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 13/25

Troubleshooting

Page 14: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 14/25

Procedure – Sample preparation

Page 15: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 15/25

Troubleshooting

Page 16: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 16/25

Troubleshooting

Page 17: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 17/25

Procedure – Purification of fungal str

Page 18: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 18/25

Troubleshooting

Page 19: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 19/25

BSL-1: Sterile conditio

• National Institute ofHygiene and Epidemiology(NIHE) in Hanoi, Viet Nam

• Separate room

• Seperate lab coat, gloves,glasses

• Laminar air flow

• 70% ethanol

• Autoclave• Sterilize all surfaces once a

month

Page 20: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 20/25

Troubleshooting

Page 21: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 21/25

Troubleshooting

Page 22: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 22/25

Troubleshooting

Page 23: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 23/25

Thank you

for your attentio

Page 24: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 24/25

Literature

• Aly AH, Debbab A, Proksch P (2011) Fungal endophytes: unique with great promises. Appl Microbiol Biotechnol 90: 1829-1845

• Ashour AA, Elkhayat ES, Ebel R, Edrada R, Proksch P (2007) Indthe Red Sea sponge Hyrtios erectus. ARKIVOC XV: 225-231

• Ding G, Liu S, Guo L, Zhou Y, Che Y (2008) Antifungal MetaboliteEndophytic Fungus Pestalotiopsis foedan . J Nat Prod 71: 615-618

• Elsässer B, Krohn K, Flörke U, Root N, Aust HJ, Draeger S, SchuKurtán T (2005) X-ray Structure determination, Absolute ConfiguraActivity of Phomoxanthone A. Eur J Org Chem 4563-4570

• Oikawa H, Yokota T, Sakano C, Suzuki Y, Naya A, Ichihara A (19Phytotoxins produced by Alternaria solani : Biosynthesis and Isolatcomponents. Biosci Biotechnol Biochem 62 (10): 2016-2022

• Scherlach K, Hertweck C (2009) Triggering cryptic natural productmicroorganisms. Org Biomol Chem 7: 1753-1760

• Xu J, Aly A H, Wray V, Proksch P (2011) Polyketide derivats of enPestalotiopsis sp. Isolated from the Chinese mangrove plant RhizoTet Let 52: 21-25

Page 25: Catalina _ Workshop1

7/23/2019 Catalina _ Workshop1.

http://slidepdf.com/reader/full/catalina-workshop1 25/25