Isolation, Purification, and Characterization of a Thermostable Xylanase from a Novel Strain, Paenibacillus campinasensis G1-1

Abstract

High levels of xylanase activity (143.98 IU/ml) produced by the newly isolated Paenibacillus campinasensis G1-1 were detected when it was cultivated in a synthetic medium. A thermostable xylanase, designated XynG1-1, from P. campinasensis G1-1 was purified to homogeneity by Octyl-Sepharose hydrophobic-interaction chromatography, Sephadex G75 gel-filter chromatography, and Q-Sepharose ion-exchange chromatography, consecutively. By multistep purification, the specific activity of XynG1-1 was up to 1,865.5 IU/mg with a 9.1-fold purification. The molecular mass of purified XynG1-1 was about 41.3 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequence analysis revealed that XynG1-1 containing 377 amino acids encoded by 1,134 bp genomic sequences of P. campinasensis G1-1 shared 96% homology with XylX from Paenibacillus campinasensis BL11 and 77%~78% homology with xylanases from Bacillus sp. YA-335 and Bacillus sp. 41M-1, respectively. The activity of XynG1-1 was stimulated by Ca 2+ , Ba 2+ , DTT, and βmercaptoethanol, but was inhibited by Ni 2+ , Fe 2+ , Fe 3+ , Zn 2+ , SDS, and EDTA. The purified XynG1-1 displayed a greater affinity for birchwood xylan, with an optimal temperature of 60 o C and an optimal pH of 7.5. The fact that XynG1-1 is cellulose-free, thermostable (stability at high temperature of 70 o C~80 o C), and active over a wide pH range (pH 5.0~9.0) suggests that the enzyme is potentially valuable for various industrial applications, especially for pulp bleaching pretreatment.

Purification, characterization and thermostability improvement of xylanase from Bacillus amyloliquefaciens and its application in pre-bleaching of kraft pulp

“…Inhibition of xylanase activity by SDS suggests presence of hydrophobic interactions in maintaining xylanase structure. Similar to our results, the xylanase activity of Paenibacillus campinasensis G1-1 was inhibited in presence of EDTA and SDS with increasing concentrations (Hongchen et al 2012). …”

Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production

“…After the stationary phase (72 h), the xylanase activity was gradully decreased because of hydrolysis by autologous protease in the decline phase of strain NF1. This course of xylanase degradation was also observed from Fomitopsis pinicola KMJ812 [15] and Paenibacillus campinasensis G1-1 [27] reported previously. Paenibacillus sp.…”

“…Endo-1,4-β-D-xylanase (E.C. 3.2.1.8) is an important enzyme for the degradation of the major hemicellulosic polysaccharide of lignocellulosic materials, since they hydrolyze xylopyranosyl linkages of β-1,4-xylan [27]. As the key xylan-degrading enzyme, xylanases have attracted considerable attention because of their widespread application in various industrial processes such as pulp bleaching, animal feeding, baking and brewing, wastetreating, bioenergy converting, textiles, and food industry [1,12].…”

“…Recently, Paenibacillus species is frequently isolated and identified from various sources for their ability to hydrolyze lignocellulosic materials. Many strains of Paenibacillus species, like P. campinasensis G1-1 [27] producing a thermostable xylanase, Paenibacillus terrae HPL-003 producing a new bimodular xylanase [17], and alkalophilic Paenibacillus sp. 12-11 [26] producing an alkaline xylanase, showed various application potential.…”

Xylanase Activity of Streptomyces violascences BF 3.10 on Xylan Corncobs and its Xylooligosaccharide Production

“…The decreased xylanase activity could be due to the hydrolysis by autologous protease in the decline phase as were reported previously in Paenibacillus sp. strain NF1 (Zheng, 2014), Fomitopsis pinicola KMJ812 (Shin et al, 2010), Paenibacillus campinasensis G1-1 (Zheng et al, 2012) and Streptomyces spp. SKK1-8 (Meryandini et al, 2006 ).…”