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Gene cloning, expression and characterization of a new cold-active and salt-tolerant endo-β-1,4-xylanase from marine Glaciecola mesophila KMM 241

Abstract

Although a lot of xylanases are studied, only a few xylanases from marine microorganisms have been reported. A new xylanase gene, xynA, was cloned from marine bacterium Glaciecola mesophila KMM 241. Gene xynA contains 1,272 bp and encodes a 423-amino acid xylanase precursor. The recombinant xylanase, XynA, expressed in Escherichia coli BL21 is a monomer with a molecular mass of 43 kDa. Among the characterized xylanases, XynA shares the highest identity (46%) to the xylanase from Flavobacterium sp. strain MSY2. The optimum pH and temperature for XynA is 7.0 and 30 °C. XynA retains 23% activity and 27% catalytic efficiency at 4 °C. XynA has low thermostability, remaining 20% activity after 60-min incubation at 30 °C. Its apparent melting temperature (T m) is 44.5 °C. These results indicate that XynA is a cold-active xylanase. XynA shows a high level of salt-tolerance, with the highest activity at 0.5 M NaCl and retaining 90% activity in 2.5 M NaCl. It may be the first salt-tolerant xylanase reported. XynA is a strict endo-β-1,4-xylanase with a demand of at least four sugar moieties for effective cleavage. It efficiently hydrolyzes xylo-oligosaccharides and xylan into xylobiose and xylotriose without producing xylose, suggesting its potential in xylo-oligosaccharides production.

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Acknowledgment

The work was supported by Hi-Tech Research and Development program of China (2007AA091504, 2007AA091903), COMRA Program (DYXM-115-02-2-6), and Specialized Research Fund for the Doctoral Program of Higher Education (20060422053).

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Correspondence to Xiu-Lan Chen.

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Guo, B., Chen, X., Sun, C. et al. Gene cloning, expression and characterization of a new cold-active and salt-tolerant endo-β-1,4-xylanase from marine Glaciecola mesophila KMM 241. Appl Microbiol Biotechnol 84, 1107–1115 (2009). https://doi.org/10.1007/s00253-009-2056-y

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Keywords

  • Xylanase
  • XynA
  • Cold-active
  • Salt-tolerant
  • Glaciecola mesophila
  • Marine
  • Xylan