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

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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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Authors and Affiliations

  1. Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081, People’s Republic of China

    Rui Zhang, Peilong Yang, Huoqing Huang, Tiezheng Yuan, Pengjun Shi, Kun Meng & Bin Yao

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  1. Rui Zhang
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  2. Peilong Yang
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  3. Huoqing Huang
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  4. Tiezheng Yuan
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  7. Bin Yao
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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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