Characterization of purified xylanase from finger millet (<Emphasis Type="Italic">Eleusine coracana-</Emphasis>Indaf 15) malt |
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Authors: | Manisseri Chithra Gudipati Muralikrishna |
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Affiliation: | (1) Department of Biochemistry and Nutrition, Central Food Technological Research Institute, Mysore, 570020, Karnataka, India |
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Abstract: | Xylanase (E.C. 3.2.1.8) was purified to apparent homogeneity from 96 h finger millet (Eleusine coracana, Indaf-15) malt by a three step purification procedure via ammonium sulphate fractionation, DEAE-cellulose ion exchange and
Sephadex G-75 gel permeation chromatographies with a recovery of 4.0% and fold purification of 60. Xylanase, having a molecular
weight of 29 ± 2 kDa was found to be monomeric on SDS-PAGE. pH optimum of the enzyme was found to be in the range of 5.0–5.5.
The activation energy was 25 kJmol−1. Xylanase showed maximum stability at 35 °C in a pH range of 5.0–6.0. K
m and V
max of purified xylanase were found to be 0.2% and 4.5 μmol min−1, respectively. Metal ions such as Ca2+, Mg2+, Mn2+, Cu2+, Fe2+, Ag2+ and Ni2+ enhanced xylanase activity at 5 mM concentration. p-chloromercuribenzoate, citric, oxalic and boric acids inhibited the enzyme in concentration dependent manner. The mode of
action of xylanase was found to be “endo” as determined by the analysis of products liberated from larchwood xylan by ESI-MS
and H1NMR. In vitro studies using Bifidobacterium and Lactobacillus sp. confirmed the prebiotic activity of the xylo-oligosaccharides. |
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Keywords: | Xylanase Finger millet Purification Xylo-oligosaccharides Prebiotic activity |
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