Fractionation and determination of proteins isolated from Cetraria islandica (L.) Ach. thallus

• Main Authors: Juodsnukytė, Jovita, Savickienė, Nijolė, Savickas, Arūnas, Žvirblytė, Gitana, Kandelinskaya, Olga, Grischenko, Helena
• Format: Conference Object
• Language: English
• Published: 2014
• Subjects: Lichens; Chemistry; Plant proteins; Analysis; Chromatography; gel; Methods; Nash; Abbott; Aldrich; Iceland; Siberia
• Online Access: http://lsmu.lvb.lt/LSMU:ELABAPDB5707791&prefLang=en_US

Lichen is a symbiotic association composed of photosynthetic partner such as green alga or cyanobacterium, and a fungal partner (Nash, 1996). Cetraria islandica (L.) Ach. (Iceland moss) is a foliaceous lichen growing mostly in central Europe, North America and Siberia (Evans, 1994). Iceland moss helps to relief dry cough, improves digestion and has antiemetic as well as immunostimulatory and anti-inflammatory effects. Cetraria islandica (L.) Ach. is a plant rich of polysaccharides, lichen acids (Ingolfsdottir et al., 1998). Cetraria islandica (L.) Ach. is also enriched with polypeptides, lichen acids and water-soluble carbohydrates. The most important of them is lichenan, (1→3,1→4) β – D – glucan (Puodžiūnienė el al., 2005; Abbott and Boraston, 2009). Proteins binding lichenan and izolichenan can be determined using Bradford dyeing method (Freysdotirr et al. 2008). For the determination of other proteins are often used Bradford and SDSPAGE assays (Vergara-Barberán et al., 2014). Aim of experiment: To determine the quantity and molecular mass of proteins isolated from Cetraria islandica (L.) Ach. thallus. Experiment tasks: 1. To separate proteins into fraction, using Size Exclusion Chromatography; 2. To estimate the quantity and molecular mass of proteins. Materials and methods: 1. Protein fractions isolated from Cetraria islandica (L.) Ach. thallus were achieved from V.F.Kuprevich Institute of Experimental Botany of Belarus Academy of Sciences. 2. Size Exclusion Chromatography was used for the proteinseparation into fractions. Size Exclusion Chromatography was performed with AKTA Purifier system, using Superdex 200 10/300 GL column. 3. The quantity of protein was measured by Bradford method using bovine serum albumin BSA (Sigma –Aldrich, USA) protein standards. 4. The molecular mass of proteins was estimated by using of Sodium dodecyl sulfate polyakrylamide gel electrophoresis SDS – PAGE. 5. Results were analyzed with MS Excel. [.].