Symbiotic Streptomyces sp. TN119 GH 11 xylanase: a new pH-stable, protease- and SDS-resistant xylanase

Abstract

A pH-stable and protease-resistant xylanase (XynB119) was identified from Streptomyces sp. TN119, a strain isolated from the gut luminal contents of longhorned beetle (Batocera horsfieldi) larvae. Using the GC TAIL-PCR method, the 1,026-bp coding gene (xynB119) with 67.3% GC content was successfully cloned and expressed in Escherichia coli. It encodes a 341-residue polypeptide with a calculated molecular mass of 35.9 kDa, including a putative 41-residue signal peptide, a catalytic domain of glycosyl hydrolase (GH) family 11, a short Gly/Pro-rich linker, and a family 2 cellulose-binding domain (CBM 2). The deduced amino acid sequence is most similar to (61.9% identity) an endo-1,4-β-xylanase from Streptomyces thermoviolaceus OPC-520. Purified recombinant XynB119 exhibited peak activity at 50 °C and pH 7.0, remained stable over a broad pH range (retaining >70% activity after incubation at pH 1.0-11.0 for 1 h at 37 °C without substrate), had strong protease resistance (retaining >90% activity after proteolytic treatment at 37 °C for 1 h) and SDS resistance (at 100 mM). These properties make XynB119 promising for application in the feed industry and valuable for basic research. Compared to r-XynB119, the r-XynB119 derivative without CBM 2 and linker region (r-XynB119d) exhibited a decreased pH stability of >25% at extreme pHs (pH 1.0-3.0 and pH 11.0-12.0).

A novel xylanase with tolerance to ethanol, salt, protease, SDS, heat, and alkali from actinomycete Lechevalieria sp. HJ3

“…100 % activity after incubation with the neutral proteases trypsin and proteinase K) [17,19,27,38] (Table 2), and these shared less than 50 % amino acid sequence identities with XynAHJ3. The K m value of xylanase XynAM6 was 2.33 mg ml -1 [27], whereas that of rXynAHJ3 was 0.8 mg ml -1 .…”

“…Glycosyl hydrolases from actinomycetes have valuable applications in various industries [2,16,17,27,38]. To date, only the taxonomic information for Lechevalieria [26,33] and genes involved in the biosynthesis of an antibiotic (rebeccamycin, accession no.…”

“…The PCR product was gel purified, ligated to pMD 18-T vector, transformed into E. coli Trans1-T1, and sequenced by Beijing Genomics Institute (Guangzhou, China). The full-length gene xynAHJ3 based on the partial endoxylanase gene was obtained using a time-saving and reducedcost GC TAIL-PCR procedure (thermal asymmetric interlaced-PCR specific for GC-rich genes) [36,38] with 4 nested insertion-specific primers (SP) ( Table 1). The annealing temperatures used for high stringency are shown in Table 1.…”

Newly Isolated <b><i>Penicillium ramulosum</i></b> N1 Is Excellent for Producing Protease-Resistant Acidophilic Xylanase

“…Several protease-resistant xylanases have been cloned from fungi. Zhou et al [2010Zhou et al [ , 2011 cloned two proteaseresistant xylanases from Streptomyces sp. TN119.…”

“…To examine resistance to different proteases, the crude enzyme was incubated at 37 ° C for 1 h with pepsin (pH 2.0), trypsin (pH 7.0), collagenase A (pH 7.0) or proteinase K (pH 7.0), and the residual enzyme activity was measured under standard conditions [Zhou et al, 2011].…”