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Microbial diversity of mangrove sediment in Shenzhen Bay and gene cloning, characterization of an isolated phytase-producing strain of SPC09 B. cereus

  • Environmental biotechnology
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Abstract

Phytases hydrolyze phytate to release inorganic phosphate, which decreases the requirement for phosphorus in fertilizers for crops and thus reduces environmental pollutants. This study analyzed microbial communities in rhizosphere sediment, collected in September 2012 from Shenzhen Bay, Guangdong, China, using high-throughput pyrosequencing; the results showed that the dominant taxonomic phyla were Chloroflexi, Firmicutes, and Proteobacteria, and the proportion of the beneficial bacteria, Bacillus, was 4.95 %. Twenty-nine culturable, phytase-producing bacteria were isolated, their phosphorus solubilization capacity was analyzed, and they were taxonomically characterized. Their phylogenetic placement was determined using 16S ribosomal RNA (rRNA) gene sequence analysis. The result shows that most of the isolates are members of the order Bacillales, although seven strains of Enterobacteriales, two strains of Pseudomonadales, and one strain of Oceanospirillales were also identified. The phytase gene was cloned from SPC09, Bacillus cereus, which showed the highest phosphorus solubilizing ability among the isolated strains. The gene encoded a primary translation product of 335 amino acids. A construct including the 1005-nt ORF fragment, Bc-phy, was transformed into Escherichia coli. The recombinant phytase was produced and purified, which revealed the temperature optima at 60 °C and pH optima at 6.5. The assessment by quantitative PCR (qPCR) showed an abundance of bacteria containing the Bc-phy gene; the level was generally higher in the mangrove forest than in the tidal flats and in surface soil compared to bottom soil, and the highest value was obtained in June. Herein, we report on the cloning, characterization, and activity of a novel phytase isolated from a mangrove system.

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Acknowledgments

This research was supported by the Guangdong Province Science and Technology Plan Project (2011B031100009), the National Marine Public Industry Research Project (201005007), The Guangdong Marine Fisheries Technology Extension Projects (A201001H02), The National Marine Public Welfare Research Project of China (201405007) and the Open Research Fund of the State Key Laboratory of Estuarine and Coastal Research (KF201209). Thanks to Prof. Jianjian Lu and Dr. Huamei Huang from ECNU for critical reading of the manuscript and to Miss Haoman Lin, Yang Dong, and Yongmei Hu for their assistance in this study.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory of Ecology and Environmental Science of Guangdong Higher Education Institutes, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou, 510631, People’s Republic of China

    Shengpeng Zhang, Shao-an Liao, Xiaoyuan Yu, Hongwu Lu, Jian-an Xian, Hui Guo & Anli Wang

  2. South China Sea Marine Engineering and Environment Institute, State Oceanic Administration, Guangzhou, 510300, People’s Republic of China

    Shengpeng Zhang & Jian Xie

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  1. Shengpeng Zhang
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Correspondence to Anli Wang.

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Zhang, S., Liao, Sa., Yu, X. et al. Microbial diversity of mangrove sediment in Shenzhen Bay and gene cloning, characterization of an isolated phytase-producing strain of SPC09 B. cereus . Appl Microbiol Biotechnol 99, 5339–5350 (2015). https://doi.org/10.1007/s00253-015-6405-8

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