Pemurnian Xilooligosakarida Hasil Hidrolisis Xilan Kulit Kopi Oleh Endo-ß-1,4-D-Xilanase XynBTN63D Menggunakan Metode Adsorpsi

Abstract

Coffee production from plantations has experienced fluctuating growth over a span of 15 years, from 2005 to 2019. The coffee industry generates a significant amount of waste in the form of coffee husks. These coffee husks consist of lignocellulosic material containing hemicellulose and other chemical compounds. The coffee husks can be utilized as a source of prebiotics through extraction to obtain xylan. Xylan extracted from coffee husks is used as a substrate for enzymatic hydrolysis using endo-β-1,4-D-xylanase enzyme, resulting in the production of xylooligosaccharides. The xylooligosaccharides derived from coffee husk xylan have a dark brown color, necessitating purification to remove any remaining impurities and separate the xylooligosaccharide components with prebiotic potential. This study aims to investigate the composition of xylooligosaccharides before and after purification using an adsorption method with column chromatography assistance. The adsorbent used is activated charcoal, and the eluent is ethanol with concentrations ranging from 10% to 50%. The results indicate that the components analyzed using TLC before purification are X2 and X3. Purification using activated charcoal as the adsorbent with 10%-50% ethanol eluent yields 32 fractions, resulting in four groups of X2-X3 fractions (at ethanol concentrations of 10%-20%). Identification of the 10%-20% ethanol fractions using TLC confirms the presence of compounds X2 and X3, while the 30%-50% ethanol fractions produce compounds X4-X5. The 10%-20% ethanol fractions, both from xylan hydrolysis using endo-β-1,4-D-xylanase pYHM-1 and endo-β-1,4-D-xylanase pESC, are selected as candidate prebiotic samples, and further identification of the components is conducted using HPLC. HPLC analysis of these fractions reveals the presence of overlapping peaks, indicating a mixture of components including X2, X3, and X4.