Bio-Precipitation of Carbonate and Phosphate Minerals Induced by the Bacterium Citrobacter freundii ZW123 in an Anaerobic Environment

Abstract

In this study, a facultative anaerobic strain isolated from marine sediments and identified as Citrobacter freundii, was used to induce the precipitation of carbonate and phosphate minerals in the laboratory under anaerobic conditions. This is the first time that the ability of C. freundii ZW123 to precipitate carbonate and phosphate minerals has been demonstrated. During the experiments, carbonic anhydrase, alkaline phosphatase and ammonium released by the bacteria not only promoted an increase in pH, but also drove the supersaturation and precipitation of carbonate and phosphate minerals. The predominant bio-mediated minerals precipitated at various Mg/Ca molar ratios were calcite, vaterite, Mg-rich calcite, monohydrocalcite and struvite. A preferred orientation towards struvite was observed. Scanning transmission electron microscopy (STEM) and elemental mapping showed the distribution of magnesium and calcium elements within Mg-rich calcite. Many organic functional groups, including C=O, C–O–C and C–O, were detected within the biominerals, and these functional groups were also identified in the associated extracellular polymeric substances (EPS). Fifteen kinds of amino acid were detected in the biotic minerals, almost identical to those of the EPS, indicating a close relationship between EPS and biominerals. Most amino acids are negatively charged and able to adsorb cations, providing an oversaturated microenvironment to facilitate mineral nucleation. The X-ray photoelectron spectroscopy (XPS) spectrum of struvite shows the presence of organic functional groups on the mineral surface, suggesting a role of the microorganism in struvite precipitation. The ZW123 bacteria provided carbon and nitrogen for the formation of the biotic minerals through their metabolism, which further emphasizes the close relationship between biominerals and the microorganisms. Thermal studies showed the enhanced thermal stability of biotic minerals, perhaps due to the participation of the bacteria ZW123. The presence of amino acids such as Asp and Glu may explain the high magnesium content of some calcites. Molecular dynamics simulations demonstrated that the morphological change and preferred orientation were likely caused by selective adsorption of EPS onto the various struvite crystal surfaces. Thus, this study shows the significant role played by C. freundii ZW123 in the bioprecipitation of carbonate and phosphate minerals and provides some insights into the processes involved.

Water Adsorption on Hydroxyapatite and Struvite as a Function of Relative Humidity: Application of BET and Freundlich Adsorption Models

“…The O 1s peaks are closely aligned between HAP and struvite, as they centered around 531.6 eV in agreement with the literature values . Struvite contains ammonium ions leading to the N 1s XPS peak, which is centered around 401.8 eV, in agreement with previous reports . HAP and struvite contain Ca 2+ and Mg 2+ , respectively, with 2p peaks centered around 347.6 and 50.8 eV, respectively. ,, The P 2p region for HAP is centered around 133.8 eV, while struvite is at 134.0 eV. ,, The ratio of Ca/P in the HAP sample used in this study was found to be 1.14 via the XPS peak ratio of Ca 2p/P 2p, which is P-enriched compared to that of previous literature reports. , This Ca/P ratio is for the surface region as it is measured using XPS, while the bulk Ca/P may differ …”

“…54,56,57 The P 2p region for HAP is centered around 133.8 eV, while struvite is at 134.0 eV. 54,55,57 The ratio of Ca/P in the HAP sample used in this study was found to be 1.14 via the XPS peak ratio of Ca 2p/P 2p, which is P-enriched compared to that of previous literature reports. 34,58 This Ca/P ratio is for the surface region as it is measured using XPS, while the bulk Ca/P may differ.…”

“…54 Struvite contains ammonium ions leading to the N 1s XPS peak, which is centered around 401.8 eV, in agreement with previous reports. 55 HAP and struvite contain Ca 2+ and Mg 2+ , respectively, with 2p peaks centered around 347.6 and 50.8 eV, respectively. 54,56,57 The P 2p region for HAP is centered around 133.8 eV, while struvite is at 134.0 eV.…”

Laboratory Experimental Study on the Formation of Authigenic Carbonates Induced by Microbes in Marine Sediments

“…Citrobacter and pseudomonas were the dominant strain at the beginning and end of the experiment, respectively. The function of formation of carbonates induced by Citrobacter was reported by previous literature [18]. Ma et al [39] injected Citrobacter into a bottle containing liquid medium.…”

“…The laboratory experiment related to authigenic carbonates, some study the formation duration of carbonate minerals under cold seep conditions [17], and some study the relationship between biominerals and the microbes [18]. However, there are few researches on the formation mechanism of carbonate minerals in marine sediments.…”