Paecilomyces variotii xylanase production, purification and characterization with antioxidant xylo-oligosaccharides production

Abstract

Paecilomyces variotii xylanase was, produced in stirred tank bioreactor with yield of 760 U/mL and purified using 70% ammonium sulfate precipitation and ultra-filtration causing 3.29-fold purification with 34.47% activity recovery. The enzyme purity was analyzed on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) confirming its monomeric nature as single band at 32 KDa. Zymography showed xylan hydrolysis activity at the same band. The purified enzyme had optimum activity at 60 °C and pH 5.0. The pH stability range was 5–9 and the temperature stability was up 70 °C. Fe2+and Fe3+ exhibited inhibition of xylanase enzyme while Cu2+, Ca2+, Mg2+ and Mn2+ stimulated its activity. Mercaptoethanol stimulated its activity; however, Na2-EDTA and SDS inhibited its activity. The purified xylanase could hydrolyze beechwood xylan but not carboxymethyl cellulose (CMC), avicel or soluble starch. Paecilomyces variotii xylanase Km and Vmax for beechwood were determined to be 3.33 mg/mL and 5555 U/mg, respectively. The produced xylanase enzyme applied on beech xylan resulted in different types of XOS. The antioxidant activity of xylo-oligosaccharides increased from 15.22 to 70.57% when the extract concentration was increased from 0.1 to 1.5 mg/mL. The enzyme characteristics and kinetic parameters indicated its high efficiency in the hydrolysis of xylan and its potential effectiveness in lignocellulosic hydrolysis and other industrial application. It also suggests the potential of xylanase enzyme for production of XOS from biomass which are useful in food and pharmaceutical industries.

Three-Step Purification and Characterization of Organic Solvent-Tolerant and Alkali-Thermo-Tolerant Xylanase from Bacillus paramycoides T4 [MN370035]

“…retained full activity in the presence of EDTA, DTT, PMSF, and β-mercaptoethanol. Contrastingly, some other works reported that EDTA [38,53] and DTT [49] slightly inhibit the xylanase activity of Bacillus sp. In the presence of EDTA, the enzyme's activity is inhibited, demonstrating that xylanase contains any form of metal ion in its active site.…”

“…The activity pattern of pure enzymes is confirmed by the fact that increasing temperature increases the rate of an enzymatic reaction, but at temperatures around the optimal value, enzyme denaturation occurs, resulting in inactivation. However, some xylanases showed higher-temperature optima [35][36][37][38][39]. Its industrial application would require significant enzyme stability at higher temperatures.…”

“…Mamoo et al [47] and Lv et al [48] have also reported that Co 2+ , Ca 2+ , Mg 2+ , Zn 2+ ,and Fe 2+ ions highly activate the xylanase activity. The stimu-latory effects of metal ions on enzyme activity could be owing to a change in the enzyme's structure [37,38]. Others such as Ni 2+ , Mn 2+ , and Na 2+ all decreased xylanase activity marginally while Cu 2+ and Hg 2+ repressed the activity highly.…”

“…Similarly, Lv et al [48] also reported that alkaliphilic xylanase from a microbial community EMSD5 exhibited a stimulatory effect on enzyme activity and stability in the presence of nonionic detergents such as Triton X-100 and Tween-20 and was slightly inhibited in the presence of SDS at a 0.1% concentration. Meanwhile, xylanase from alkalophilic bacteria Bacillus pumilus, Trichoderma inhamatum, and Paecilomyces variotii lost their total activity in the presence of SDS [23,31,38]. In another study, xylanase of Simplicillium obclavatum showed maximum enzyme activity in the presence of SDS while inhibiting its activity in the presence of Triton X-100 [43].…”

Preparation and nutritional properties of xylooligosaccharide from agricultural and forestry byproducts: A comprehensive review

“…Helveticus ); the antibacterial components of cauliflower stalkm XOS were dihydroxybenzoic acid and aspartic acid ( 99 ). Abdella et al reported that after the xylanase produced by Paecilomyces wallichii was applied to beech xylan to produce different types of XOS; when the extract concentration was from 0.1 to 1.5 mg/mL, the antioxidant activity of XOS increased from 15.22 to 70.57% ( 100 ). XOS from oil palm empty fruit bunch hemicellulose produced by xylanase from Thermomyces cyanobacterium hydrolysis was composed of xylotriose and xylobiose.…”