A new bi-modular endo-β-1,4-xylanase KRICT PX-3 from whole genome sequence of Paenibacillus terrae HPL-003

Abstract

A new bi-modular, wide pH spectrum and highly active xylanase KRICT PX3 (JF320814) isolated from Paenibacillus terrae HPL-003 (KCTC11987BP) has been cloned and expressed in Escherichia coli. Purified recombinant xylanase KRICT PX-3 (1,620 bp, 540aa, NCBI accession number JF320814) showed highly active at 55°C in pH 4.0-11.0, and stability for at least 24h at 50°C, and exhibited Km and Vmax of 0.2mg/mL and 153.8 U/mg on birchwood xylan. Most common ions did not affect the enzyme activity at 1mM concentration. This enzyme could belong to glycoside hydrolase family 10 because hydrolyzed glucuronoxylan and arabinoxylan substrate to xylobiose, xylotriose, and some traces of xylose as hydrolysis products. Model 3-D structure was composed of two domains, the catalytic domain of a (β/α)8 barrel fold while the small domain probably functions as a xylan binding domain, and the two domains are connected by a flexible linker peptide (PPLAIEKDIPSL). However, sequence alignment between xylan-binding module in this xylanase KRICT PX3 and CBM22 showed 21% of identity and 35% of similarity. This xylanase structure showed a distinctive group of enzyme cluster separately from the rest of GH10 xylanases, and seems to constitute a new type of xylanases.

Direct cloning, expression of a thermostable xylanase gene from the metagenomic DNA of cow dung compost and enzymatic production of xylooligosaccharides from corncob

“…N16-5 (83 % identity), which is optimal at 70°C and pH 7.0, Xyn10CD18 showed higher thermostability and higher acid resistance (Zhang et al2010). Similar strong specificity toward birchwood xylan and oat spelt xylan was also verified for xylanases from Paenibacillus terrae HPL-003 (Song et al 2014), Paenibacillus sp. In addition, the favorable properties of Xyn10CD18 could make it suitable for industrial applications.…”

Purification, Identification, and Characterization of an Endo-1,4-β-Xylanase from Wheat Malt

“…However, there are few reports in this field. Endo-1,4-β-xylanase is a highly diversified enzyme [13] in various aspects such as structural domains, biochemical properties and catalytic characteristics. Many studies have been carried out to improve the enzyme stability through genetic recombination [14] and modification.…”

Cloning and expression of a broad pH stable GH11 xylanase from Aspergillus niger C71 / Aspergillus niger C71 den geniş pH aralığında dengeli olan GH11 xylanaz klonlaması ve ekspresyonu

“…Recently, many xylanases have been isolated from fungi, bacteria, actinomycetes and archaea, and the genes of xylnases have been cloned and sequenced [3][4][5]. To date, microbial xylanases have been classified into nine glycoside hydrolase (GH) families: 5,8,10,11,16,26,30,43 and 62 based on the similarity of the catalytic domain sequences (www.CAZy.org).…”

“…To date, microbial xylanases have been classified into nine glycoside hydrolase (GH) families: 5,8,10,11,16,26,30,43 and 62 based on the similarity of the catalytic domain sequences (www.CAZy.org). The true β-1,4-acting xylanases with a unique catalytic domain are restricted to GH families 5, 8, 10, 11 and 30.…”