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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

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Overview

• Introduction

• General methods for isolation of derived fungi

• Workup of fractions

• Troubleshooting

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Motivation

• Discovery of Penicillin1928, by A.Flemming

Modern era of

antibiotic discovery• Fungi were now seen

as potential medicinefactories

Fig.1: Alexander

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Endophytic fungi

• Reside in the internal tissue ofplants

• Secondary metabolites utile

for – Plants

• second messengers

• protection against herbivores

 – Humans• Paclitaxel

Fig

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Marine endophytes

• Extreme habitat

adaptaion necessary

• High chemical diversity promising potential as

sources of new andbioactive natural products

Fig.3: Mang

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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

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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

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Isolation of endophyt

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Fermentation

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Extraction from rice med

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Isolation and structure elucof bioactive secondary meta

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Main literature

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Troubleshooting

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Procedure – Sample preparation

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Troubleshooting

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Troubleshooting

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Procedure – Purification of fungal str

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Troubleshooting

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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

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Troubleshooting

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Troubleshooting

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Troubleshooting

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Thank you

for your attentio

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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

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