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Undersökning av den naturliga cellulasinhibitorn från lagerbärsarten Myrica pensylvanica
1999 (Swedish)Independent thesis Basic level (professional degree)Student thesis
Abstract [sv]

Karboximetylcellulosa, CMC, ökar viskositeten hos vattenlösningar. CMC-innehållande produkter kontamineras emellanåt av cellulaser från svampar och bakterier. Lagerbärsarten Myrica pensylvanica innehåller en effektiv cellulasinhiberande substans, troligen en kondenserad tannin eller möjligen en anthocyanin. Syftet med mitt examensarbete var att isolera och identifiera den cellulasinhiberande substansen. Inhibitorsubstansen renades genom syrafällning (koncentrerad saltsyra) och undersöktes med hjälp av kromatografi och spektroskopi. För all kromatografi användes en HPLC-utrustning. Tre kolonner provades: En katjonbytarkolonn med fosfatbuffert och metanol som lösningsmedel, en Sephadex LH20-kolonn och en C18-kolonn med olika lösningsmedelssystem innehållande vatten, metanol och ättiksyra eller myrsyra. Endast med C18-kolonnen kunde fraktioner med inhibitorsubstans elueras. För identifiering av substansen användes papperskromatografi, UV/VIS-spektroskopi och NMR-spektra. Papperskromatogrammen visade två, ej helt separerade, olikfärgade band. UV/VIS-spektroskopin bekräftade att färgämnet i inhibitorn är en anthocyanidin, troligen cyanidin eller 3,7,8,3',4'-pentahydroxyflavylium eller båda. Samtliga NMR-spektra innehöll signaler i det aromatiska området, men var inte tillräckligt detaljerade för identifiering av substansen. För att kunna identifiera inhibitorsubstansen krävs en effektiv metod att separera de kondenserade tanninerna. Dessutom krävs spektra från högfälts-NMR.

Abstract [en]

Carboxymethylcellulose, CMC, increases the viscosity of water solutions. CMC-containing products are occasionally contaminated by cellulases from fungi and bacteria. The bayberry species Myrica pensylvanica contains an effective cellulase inhibitor, probably a condensed tannin or possibly an anthocyanine. The aim of my project has been to isolate and identify the cellulase inhibitory substance. The inhibitory substance was purified by means of acid precipitation (concentrated hydrochloric acid), and was characterised by chromatographic and spectroscopic methods. Chromatography was carried out using an HPLC instrument with three different columns: One cation exchange column with phosphate buffer/methanol as solvent, one Sephadex LH20 column and one C18 column with different solvent systems containing water, methanol and acetic acid or formic acid. Fractions containing inhibitor were eluted only from the C18 column. Paper chromatography, UV/VIS spectroscopy and NMR spectroscopy were used to identify the substance. The paper chromatograms showed two bands, differently coloured, and not entirely separated. UV/VIS spectroscopy confirmed that the red pigment in the inhibitor is an anthocyanidin, probably cyanidin or 3,7,8,3',4'-penta-hydroxyflavylium or both. All NMR spectra contained signals in the aromatic region, but they were insufficiently detailed for identification of the substance. Identification of the inhibitory substance would require an efficient method of separating the condensed tannins and in addition high field NMR spectra.

Place, publisher, year, edition, pages
1999. , 51 p.
Identifiers
URN: urn:nbn:se:kau:diva-52638Local ID: KEM-9OAI: oai:DiVA.org:kau-52638DiVA: diva2:1101165
Subject / course
Chemistry
Available from: 2017-05-29 Created: 2017-05-29

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