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

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.

Diverse Profiles of AI-1 Type Quorum Sensing Molecules in Cultivable Bacteria from the Mangrove (Kandelia obovata) Rhizosphere Environment

“…In the present study, we focused on K. obovata and found that prokaryotic communities in mangrove roots were dominated by Proteobacteria, Chloroflexi, Bacteroidetes (previously Cytophaga–Flexibacter–Bacteroides), Planctomycetes, and Actinobacteria ( Supplementary Figure S1A ). The observation was consistent with previous reports (Andreote et al, 2012; Hong et al, 2015; Zhang et al, 2015). Among the Proteobacteria, Gamma-proteobacteria, Delta-proteobacteria, and Alpha-proteobacteria were the three most abundant classes in our sampling sites ( Supplementary Figure S1B ), in agreement with previous findings in Aegiceras corniculatum , Avicennia marina , and Sonneratia caseolaris (Yang et al, 2014; Alzubaidy et al, 2016; Chen et al, 2016).…”

Isolation and Characterization of Phosphorus Solubilizing Bacteria With Multiple Phosphorus Sources Utilizing Capability and Their Potential for Lead Immobilization in Soil

“…Some PSB present higher P solubilizing ability than that of A. pittii gp-1, such as Pseudomonas trivialis BIHB 745 for Ca 3 (PO 4 ) 2 (827 mg/L) (Vyas and Gulati, 2009), Serratia marcescens RP8 for Ca 3 (PO 4 ) 2 (974 mg/L) (Misra et al, 2012), Acinetobacter sp. ASL12 for Ca 3 (PO 4 ) 2 (717 mg/L) (Liu et al, 2014), Bacillus cereus SPC09 for phytate (579.01 mg/L) (Zhang et al, 2015), Saccharomyces cerevisiae CICIMY008 for phytate (93 mg/L) (Chen et al, 2016), Burkholderia cepacia B116 for phytate (52.7 mg/L) (Oliveira et al, 2009). However, the multiple P sources utilizing ability promises gp-1 itself easily get used to environment, and therefore exhibits great potential for phosphate chemical industry and agro-ecosystems.…”

“…Phosphorus solubilizing microbes especially PSB are widely distributed in soils, freshwater, seawater, and sediments (Liu et al, 2014(Liu et al, , 2017Zhang et al, 2015), and responsible for the cycling of insoluble P to soluble PO 4 3− ion. Numerous researches have concentrated on the screening of highly efficient PSB, and most PSB are Gram-negative bacteria and belong to Pseudomonas (Misra et al, 2012;Oteino et al, 2015), Acinetobacter (Liu et al, 2014(Liu et al, , 2015, Pantoea and Enterobacter (Park et al, 2011;Chen and Liu, 2019), and some PSB are Gram-positive bacteria belonging to Bacillus (Hanif et al, 2015;Wang et al, 2017).…”

Phosphate-Solubilizing Fungi and Alkaline Phosphatase Trigger the P Solubilization During the Co-composting of Sorghum Straw Residues With Burkina Faso Phosphate Rock

“…However, since pqqA, pqqB, pqqC, pqqD, and pqqE genes are conserved and arranged in that order in the pqq operon (pqqABCDE) (Shen et al, 2012), we selected the pqqB and pqqE genes to study the mPSB. Given the substantial wide distribution of PSB in natural ecosystems including soils, freshwater, seawater, and sediments (Zhang et al, 2015;Liang et al, 2020;Wan et al, 2020), we cannot completely rule out a possible contribution of phosphate solubilizing bacteria in the increase of P fractions in our compost treatments. The moderate but positive correlation of 16S bacteria with the labile-P supports the possibility of bacteria to intervene in the P solubilization in the composts.…”