Abstract
A glycosyl hydrolase family 10 endoxylanase from Bacillus sp. HJ14 was grouped in a separated cluster with another six Bacillus endoxylanases which have not been characterized. These Bacillus endoxylanases showed less than 52 % amino acid sequence identity with other endoxylanases and far distance with endoxylanases from most microorganisms. Signal peptide was not detected in the endoxylanase. The endoxylanase was expressed in Escherichia coli BL21 (DE3), and the purified recombinant enzyme (rXynAHJ14) was characterized. rXynAHJ14 was apparent optimal at 62.5 °C and pH 6.5 and retained more than 55 % of the maximum activity when assayed at 40–75 °C, 23 % at 20 °C, 16 % at 85 °C, and even 8 % at 0 °C. Half-lives of the enzyme were more than 60 min, approximately 25 and 4 min at 70, 75, and 80 °C, respectively. The enzyme exhibited more than 62 % xylanase activity and stability at the concentration of 3–30 % (w/v) NaCl. No xylanase activity was lost after incubation of the purified rXynAHJ14 with trypsin and proteinase K at 37 °C for 60 min. Different components of oligosaccharides were detected in the time-course hydrolysis of beechwood xylan by the enzyme. During the simulated intestinal digestion phase in vitro, 11.5–19.0, 15.3–19.0, 21.9–27.7, and 28.2–31.2 μmol/mL reducing sugar were released by the purified rXynAHJ14 from soybean meal, wheat bran, beechwood xylan, and rapeseed meal, respectively. The endoxylanase might be an alternative for potential applications in the processing of sea food and saline food and in aquaculture as agastric fish feed additive.
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Acknowledgments
This work was supported by the Key Technologies Research and Development Program of China (2013BAD10B01), National Natural Science Foundation of China (No. 31260215), and Science Research Foundation of Yunnan Provincial Education Committee (No. 2012Y166). We thank Prof. James F. Preston from the University of Florida for his kind gifts (methylglucuronoxylose, methylglucuronoxylobiose, methylglucuronoxylotriose, and methylglucuronoxylotetraose).
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Junpei Zhou and Qian Wu contributed equally to this work.
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Zhou, J., Wu, Q., Zhang, R. et al. A thermo-halo-tolerant and proteinase-resistant endoxylanase from Bacillus sp. HJ14. Folia Microbiol 59, 423–431 (2014). https://doi.org/10.1007/s12223-014-0316-4
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DOI: https://doi.org/10.1007/s12223-014-0316-4