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Molecular and biochemical characterization of a new alkaline β-propeller phytase from the insect symbiotic bacterium Janthinobacterium sp. TN115

Abstract

A phytase-encoding gene (phyA115) was cloned from Janthinobacterium sp. TN115, a symbiotic bacterial strain isolated from the gut contents of Batocera horsfieldi larvae (Coleoptera: Cerambycidae), and expressed in Escherichia coli. The 1,884-bp full-length gene encodes a 28-residue putative signal peptide and a 599-residue mature protein with a calculated mass of 64 kDa. The deduced PhyA115 shares low identity with known sequences (47% at most) and contains an N-terminal incomplete domain (residues 29–297; domain N) and a typical β-propeller phytase domain at the C terminus (residues 298–627; domain C). Distinct from other β-propeller phytases that have neutral pH optima (pH 6.0–7.5), purified recombinant PhyA115 exhibits maximal activity at pH 8.5 and 45°C in the presence of 1 mM Ca2+ and is highly active over a wider pH range (pH 6.0–9.0). These results indicate that PhyA115 is a β-propeller phytase that has application potential in aquaculture feed. To our knowledge, this is the first report of cloning of a phytase gene from the symbiotic microbes of an insect digestive tract and from the genus Janthinobacterium. The N-terminal incomplete domain is found to have no phytase activity but can influence the pH property of PhyA115.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31001025), the Key Program of Transgenic Plant Breeding (2008ZX08011-005), and the Earmarked Fund for Modern Agro-industry Technology Research System (NYCYTX-42-G2-05).

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Correspondence to Bin Yao.

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Zhang, R., Yang, P., Huang, H. et al. Molecular and biochemical characterization of a new alkaline β-propeller phytase from the insect symbiotic bacterium Janthinobacterium sp. TN115. Appl Microbiol Biotechnol 92, 317–325 (2011). https://doi.org/10.1007/s00253-011-3309-0

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Keywords

  • β-Propeller phytase
  • Janthinobacterium sp. TN115
  • Alkaline phytase
  • Batocera horsfieldi (Coleoptera: Cerambycidae)