Production, Purification, and Characterization of Thermostable Alkaline Xylanase From Anoxybacillus kamchatkensis NASTPD13

Abstract

Anoxybacillus kamchatkensis NASTPD13 used herein as a source for thermostable alkaline xylanase were isolated from Paudwar Hot Springs, Nepal. NASTPD13 cultured at 60°C, pH 7 and in presence of inorganic (ammonium sulfate) or organic (yeast extract) nitrogen sources, produced maximum xylanase enzyme. Xylanase production in the cultures was monitored by following the ability of culture media to hydrolyze beech wood xylan producing xylooligosaccharide and xylose by thin layer chromatography (TLC). The extracellular xylanase was isolated from optimized A. kamchatkensis NASTPD13 cultures by ammonium sulfate (80%) precipitation; the enriched xylanase preparation was dialyzed and purified using Sephadex G100 column chromatography. The purified xylanaseshowed 11-fold enrichment with a specific activity of 33 U/mg and molecular weight were37 kDa based on SDS-PAGE and PAGE-Zymography. The optimum pH and temperature of purified xylanase was 9.0 and 65°C respectively retainingmore than 50% of its maximal activity over a broad range of pH (6–9) and temperature (30–65°C). With beech wood xylan, the enzyme showed Km 0.7 mg/ml and Vmax 66.64 μM/min/mg The xylanase described herein is a secretory enzyme produced in large quantities by NASTPD13 and is a novel thermostable, alkaline xylanase with potential biotechnological applications.

A detailed overview of xylanases: an emerging biomolecule for current and future prospective

“…Usually, it has been observed that xylanase purification was performed by the multi-step process where the concentration of protein using ammonium acetate/ TCA/acetone precipitation or ultracentrifugation was followed by a single step or series of chromatography techniques. Yadav et al (2018) demonstrated the purification and characterization of extracellular xylanase obtained from A. kamchatkensis NASTPD13 cultures. The crude xylanase was subjected to ammonium sulfate (80%) precipitation followed by dialysis.…”

Augmenting Pentose Utilization and Ethanol Production of Native Saccharomyces cerevisiae LN Using Medium Engineering and Response Surface Methodology

“…Second generation bioethanol, produced by fermentation of sugars present in lignocellulosic biomass is a promising alternative fuel, due to the declining availability of fossil fuels and their negative impact on the environment (Cai et al, 2012 ; Lin et al, 2012 ; Yadav et al, 2018 ). Saccharomyces cerevisiae , widely used in starch and sucrose based 1st generation bioethanol production, is believed to be the most appropriate candidate for lignocellulosic bioethanol production due to its highly efficient hexose fermentation under anaerobic conditions, high tolerance to both ethanol and inhibitory compounds present in lignocellulosic hydrolysates.…”

Development of novel economical methodologies for zymographic analysis of purified xylano–pectinolytic enzymes using agrowaste‐based substrates

“…Kumar et al supplemented gel with Birchwood xylan for zymographic analysis of xylanase enzyme. Yadav et al copolymerized Beechwood xylan (0.2% wt/vol) in PAGE gel and performed zymography after electrophoresis. Subsequent staining with Congo red (0.1% wt/vol) and destaining with NaCl (1 M) revealed the zymographic band corresponding to 37 kDa.…”

“…Kumar et al28 supplemented gel with Birchwood xylan for zymographic analysis of xylanase enzyme. Yadav et al29 cop-…”