catalina _ workshop1
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7/23/2019 Catalina _ Workshop1.
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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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