Selectivity in Enzymatic Phosphorus Recycling from Biopolymers: Isotope Effect, Reactivity Kinetics, and Molecular Docking with Fungal and Plant Phosphatases.

Among ubiquitous phosphorus (P) reserves in environmental matrices are ribonucleic acid (RNA) and polyphosphate (polyP), which are, respectively, organic and inorganic P-containing biopolymers. Relevant to P recycling from these biopolymers, much remains unknown about the kinetics and mechanisms of different acid phosphatases (APs) secreted by plants and soil microorganisms. Here we investigated RNA and polyP dephosphorylation by two common APs, a plant purple AP (PAP) from sweet potato and a fungal phytase from Aspergillus niger. Trends of δ18O values in released orthophosphate during each enzyme-catalyzed reaction in 18O-water implied a different extent of reactivity. Subsequent enzyme kinetics experiments revealed that A. niger phytase had 10-fold higher maximum rate for polyP dephosphorylation than the sweet potato PAP, whereas the sweet potato PAP dephosphorylated RNA at a 6-fold faster rate than A. niger phytase. Both enzymes had up to 3 orders of magnitude lower reactivity for RNA than for polyP. We determined a combined phosphodiesterase-monoesterase mechanism for RNA and terminal phosphatase mechanism for polyP using high-resolution mass spectrometry and 31P nuclear magnetic resonance, respectively. Molecular modeling with eight plant and fungal AP structures predicted substrate binding interactions consistent with the relative reactivity kinetics. Our findings implied a hierarchy in enzymatic P recycling from P-polymers by phosphatases from different biological origins, thereby influencing the relatively longer residence time of RNA versus polyP in environmental matrices. This research further sheds light on engineering strategies to enhance enzymatic recycling of biopolymer-derived P, in addition to advancing environmental predictions of this P recycling by plants and microorganisms.

How dam construction affects the activity of alkaline phosphatases in reservoir sediments: A study of two highly regulated rivers.

Dam construction causes phosphorus (P) accumulation in reservoir sediments and significantly affects the generation of available P. However, the effect of dam construction on the activity of sediment alkaline phosphatase (ALP), which is encoded by the bacterial phoD gene and participates in P mineralization, in river sediments remains unclear. Here, we investigated the ALP activities in 78 sediment samples collected from the cascade reservoir regions located in the Lancang River and the Jinsha River, two highly regulated rivers in southwestern China. The abundance and community composition of phoD-harboring bacteria were determined based on the phoD gene using quantitative real-time PCR and MiSeq sequencing, respectively. Comparison of control and affected sites indicated that dam construction significantly increased sediment ALP activity in both rivers. The abundances of phoD-harboring bacteria increased and their community compositions varied in response to dam construction; the relative abundances of the dominant genera Methylobacterium and Bradyrhizobium were particularly higher in affected site than control site. Co-occurrence network analyses revealed much higher network connectivity and relative abundances of keystone species in affected sites. Some microbial factors including phoD-harboring bacterial abundances, network clustering coefficients, and relative abundance of keystone species were positively correlated with ALP activity. The relative abundance of keystone species was identified as the most important microbial factor contributing to variation in ALP activity based on structural equation modeling analysis. These findings enhance our understanding of how dam construction affects the functions of phoD-harboring bacteria and their role in the P biogeochemical cycle in highly regulated rivers.

Capability of Penicillium oxalicum y2 to release phosphate from different insoluble phosphorus sources and soil.

Due to insufficient amount of soluble phosphate and poor persistence of traditional chemical phosphate fertilizers in agricultural soils, the eco-friendly and sustainable phosphorus sources for crops are urgently required. The efficient phosphate-releasing fungal strain designated y2 was isolated and identified by the internal transcribed spacer of rDNA as Penicillium oxalicum y2. When lecithin, Ca3(PO4)2, or ground phosphate rock were separately used as sole phosphorus source, different phosphate-releasing modes were observed. The strain y2 was able to release as high as 2090 mg/L soluble phosphate within 12 days of incubation with Ca3(PO4)2 as sole phosphorus source. In the culture solution, high concentration of oxalic, citric, and malic acids and high phosphatase activity were detected. The organic acids contributed to solubilizing inorganic phosphate sources, while phosphatase was in charge of the mineralization of organic phosphorus lecithin. Afterwards, the fungus culture was applied to the soil with rape growing. During 50 days of incubation, the soil's available phosphate concentration increased by three times compared with the control, the dry weight of rape increased by 78.73%, and the root length increased by 38.79%. The results illustrated that P. oxalicum y2 possessed both abilities of solubilizing inorganic phosphorus and mineralizing organic phosphorus, which have great potential application in providing biofertilizer for modern agriculture.

In-situ organic phosphorus mineralization in sediments in coastal wetlands with different flooding periods in the Yellow River Delta, China.

In-situ incubation experiments were performed in typical tidal flooding wetlands and seasonal flooding wetlands in the Yellow River Delta of China to investigate sediment organic phosphorus (OP) mineralization and its influencing factors. The results showed that the sediment net P mineralization rate (RNPM) exhibited consistent seasonal variations in both wetlands, and it was more stable in the tidal flooding wetlands than in the seasonal flooding wetlands. Sediment P mineralization was greatly influenced by plant uptake and flooding erosion, and the freshwater input by flow-sediment regulation replenished the inorganic phosphorus (IP) pool in the wetland sediments. The OP, IP and total P in the sediments of the tidal flooding wetlands were in a state of dynamic equilibrium throughout the plant growing season, and plant uptake peaked during the period from August to September. In the seasonal flooding wetlands, rainfall and flow-sediment regulation were the key factors influencing the conversion between OP and IP. Besides sediment salinity and pH, microbial biomass and enzyme activities were also the key factors influencing the sediment RNPM in both wetlands. The findings of this study indicated that flooding frequencies and salinity could highly impact sediment P mineralization, and that the IP levels in sediments might be influenced by wetland hydrology and salinity.

The diversity and abundance of phytase genes (ß-propeller phytases) in bacterial communities of the maize rhizosphere.

UNLABELLED: The ecology of microbial communities associated with organic phosphorus (P) mineralization in soils is still understudied. Here, we assessed the abundance and diversity of bacteria harbouring genes encoding ß-propeller phytases (BPP) in the rhizosphere of traditional and transgenic maize cultivated in two Brazilian soils. We found a soil-dependent effect towards a higher abundance of phytase genes in the rhizosphere, and an absence of any impact of plant genotype. Phylogenetic analyses indicated members of the genera Pseudomonas, Caulobacter, Idiomarina and Maricaulis, close to 'uncultured bacteria', to constitute the dominant bacteria hosting this gene. The results obtained validate a methodology to target bacteria that are involved in the organic P cycle, and depict the responsiveness of such bacteria to the rhizosphere, albeit in dependency of the soil in which maize is cultivated. The data also identified the major bacterial groups that are associated with the organic P mineralization function. SIGNIFICANCE AND IMPACT OF THE STUDY: Micro-organisms play a key role in nutrient balance in soil ecosystems that are essential to life on the planet. However, some processes such as organic phosphorus mineralization, an important source of phosphorus supply in soil, is poorly studied mainly due the absence of an efficient methodology to assess the phytase-producing micro-organisms. In this study, a method to assess beta-propeller phytase (BPP)-carrying bacteria in soil was validated. This method may contribute to the knowledge of how these micro-organisms behave in the environment and contribute for plant growth promotion.