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Microbially induced calcite precipitation using Bacillus velezensis with guar gum.

ABSTRACT: Mineral precipitation via microbial activity is a well-known process with applications in various fields. This relevance of microbially induced calcite precipitation (MICP) has pushed researchers to explore various naturally occurring MICP capable bacterial strains. The present study was performed to explore the efficiency of microbially induced calcite precipitation (MICP) via locally isolated bacterial strains and role of guar gum, which is a naturally occurring polymer, on the MICP process. The strains were isolated from local soil and screened for urease activity Further, the urease positive strain was subjected to urea and calcium chloride based medium to investigate the efficacy of isolated strain for microbial induced precipitation. Among screened isolates, the soil bacterium that showed urease positive behaviour and precipitated calcium carbonate was subjected to 16S rRNA gene sequencing. This strain was identified as Bacillus velezensis. Guar gum-a natural polymer, was used as a sole carbon source to enhance the MICP process. It was observed that the isolated strain was able to breakdown the guar gum into simple sugars resulting in two-fold increase in calcium carbonate precipitate. Major bio-chemical activities of isolated strain pertaining to MICP such as ammonium ion concentration, pH profiling, and total reducing sugar with time were explored under four different concentrations of guar gum (0.25%, 0.5%, 0.75% and 1% w/v). Maximum ammonium ion concentration (17.5 ?g/ml) and increased pH was observed with 1% guar gum supplementation, which confirms augmented MICP activity of the bacterial strain. Microstructural analysis of microbial precipitation was performed using scanning electron microscopy (SEM) and X-ray diffraction (XRD) techniques, which confirmed the presence of calcium carbonate in different phases. Further, XRD and SEM based studies corroborated that guar gum supplemented media showed significant increase in stable calcite phase as compared to media without guar gum supplementation. Significant diverse group of nitrogenous compounds were observed in guar gum supplemented medium when subjected to Gas Chromatography-Mass spectrometry (GC-MS) profiling.

SUBMITTER: Dikshit R

PROVIDER: S-EPMC7423064 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Microbially induced calcite precipitation using Bacillus velezensis with guar gum.

Dikshit Rashmi R Jain Animesh A Dey Arjun A Kumar Aloke A

PloS one 20200812 8

Mineral precipitation via microbial activity is a well-known process with applications in various fields. This relevance of microbially induced calcite precipitation (MICP) has pushed researchers to explore various naturally occurring MICP capable bacterial strains. The present study was performed to explore the efficiency of microbially induced calcite precipitation (MICP) via locally isolated bacterial strains and role of guar gum, which is a naturally occurring polymer, on the MICP process. T ...[more]

PMID: 32785276

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Project description:BackgroundMicrobially induced calcite precipitation (MICP) is an ancient property of bacteria, which has recently gained considerable attention for biotechnological applications. It occurs as a by-product of bacterial metabolism and involves a combination of chemical changes in the extracellular environment, e.g. pH increase, and presence of nucleation sites on the cell surface or extracellular substances produced by the bacteria. However, the molecular mechanisms underpinning MICP and the interplay between the contributing factors remain poorly understood, thus placing barriers to the full biotechnological and synthetic biology exploitation of bacterial biomineralisation.ResultsIn this study, we adopted a bottom-up approach of systematically engineering Bacillus subtilis, which has no detectable intrinsic MICP activity, for biomineralisation. We showed that heterologous production of urease can induce MICP by local increases in extracellular pH, and this can be enhanced by co-expression of urease accessory genes for urea and nickel uptake, depending on environmental conditions. MICP can be strongly enhanced by biofilm-promoting conditions, which appeared to be mainly driven by production of exopolysaccharide, while the protein component of the biofilm matrix was dispensable. Attempts to modulate the cell surface charge of B. subtilis had surprisingly minor effects, and our results suggest this organism may intrinsically have a very negative cell surface, potentially predisposing it for MICP activity.ConclusionsOur findings give insights into the molecular mechanisms driving MICP in an application-relevant chassis organism and the genetic elements that can be used to engineer de novo or enhanced biomineralisation. This study also highlights mutual influences between the genetic drivers and the chemical composition of the surrounding environment in determining the speed, spatial distribution and resulting mineral crystals of MICP. Taken together, these data pave the way for future rational design of synthetic precipitator strains optimised for specific applications.

Project description:Microbial-Induced Calcite Precipitation (MICP) stimulates soil microbiota to induce a cementation of the soil matrix. Urea, calcium and simple carbon nutrients are supplied to produce carbonates via urea hydrolysis and induce the precipitation of the mineral calcite. Calcium chloride (CaCl2) is typically used as a source for calcium, but basic silicate rocks and other materials have been investigated as alternatives. Weathering of calcium-rich silicate rocks (e.g., basalt and dolerite) releases calcium, magnesium and iron; this process is associated with sequestration of atmospheric CO2 and formation of pedogenic carbonates. We investigated atmospheric carbon fluxes of a MICP treated sandy soil using CaCl2 and dolerite fines applied on the soil surface as sources for calcium. Soil-atmosphere carbon fluxes were monitored over 2 months and determined with an infrared gas analyser connected to a soil chamber. Soil inorganic carbon content and isotopic composition were determined with isotope-ratio mass spectrometry. In addition, soil-atmosphere CO2 fluxes during chemical weathering of dolerite fines were investigated in incubation experiments with gas chromatography. Larger CO2 emissions resulted from the application of dolerite fines (116 g CO2-C m-2) compared to CaCl2 (79 g CO2-C m-2) but larger inorganic carbon precipitation also occurred (172.8 and 76.9 g C m-2, respectively). Normalising to the emitted carbon to precipitated carbon, the environmental carbon cost was reduced with dolerite fines (0.67) compared to the traditional MICP treatment (1.01). The carbon isotopic signature indicated pedogenic carbonates (δ13Cav = -8.2 ± 5.0‰) formed when dolerite was applied and carbon originating from urea (δ13Cav = -46.4 ± 1.0‰) precipitated when CaCl2 was used. Dolerite fines had a large but short-lived (<2 d) carbon sequestration potential, and results indicated peak CO2 emissions during MICP could be balanced optimising the application of dolerite fines.

Project description:Biomineralization plays a key role in modifying the geological properties of soil, thereby stabilizing it against wind erosion, especially in areas characterized by harsh weather and harsh soil (calcareous and arid); i.e. Arabic Gulf region. Among soil microorganisms, ureolytic bacteria are capable of modifying soil characteristics and thus, inducing biomineralization. This research investigated the occurrence and diversity of ureolytic bacteria in Qatari soils, specifically to study their acquired potential to adapt to harsh conditions exhibiting ureolytic activity. Soil samples were collected from various locations in Qatar and were used to isolate the indigenous ureolytic bacteria. It was noticed that most of the ureolytic bacteria in Qatari soil belong to the genus Bacillus mainly Bacillus cereus. Identification and differentiation of 18 ureolytic isolates were performed using MALDI-TOF MS techniques while ribotyping (16S rRNA) molecular technique was used mainly for 6 selected strains. This study not only shows the diversity of species of ureolytic bacteria in Qatari soil but also shows the diversity in their protein profiles, which confirms that bacteria have adapted well to the harsh environment. In addition, the strains were evaluated based on a newly modified screening method in this work; i.e. production of arbitrary urease activity (AUA). Thus, the strains showing the highest AUA, exhibited the highest capability to produce urease enzymes induced by urea. Analysis of calcium carbonate precipitation utilizing SEM-EDX showed that the ureolytic bacteria also play a significant role in the precipitation of minerals such as CaCO3, in the presence of urea in soil. Therefore, this research showed a high occurrence of indigenous Bacillus bacteria in Qatari soil that can perform biomineralization and thus can be helpful, if properly stimulated, in enhancing soil stabilization, and for other local applications as well, since they are adapted to these soil and weather conditions.

Project description:Soil properties are the most important factors determining the safety of civil engineering structures. One of the soil improvement methods studied, mainly under laboratory conditions, is the use of microbially induced calcite precipitation (MICP). Many factors influencing the successful application of the MICP method can be distinguished; however, one of the most important factors is the composition of the bio-cementation solution. This study aimed to propose an optimal combination of a bio-cementation solution based on carbonate precipitation, crystal types, and the comprehensive strength of fine sand after treatment. A series of laboratory tests were conducted with the urease-producing environmental strain of bacteria B. subtilis, using various combinations of cementation solutions containing precipitation precursors (H2NCONH2, C6H10CaO6, CaCl2, MgCl2). To decrease the environmental impact and increase the efficiency of MICP processed, the addition of calcium lactate (CaL) and Mg ions was evaluated. This study was conducted in Petri dishes, assuming a 14-day soil treatment period. The content of water-soluble carbonate precipitates and their mineralogical characterization, as well as their mechanical properties, were determined using a pocket penetrometer test. The studies revealed that a higher concentration of CaL and Mg in the cementation solution led to the formation of a higher amount of precipitates during the cementation process. However, the crystal forms were not limited to stable forms, such as calcite, aragonite, (Ca, Mg)-calcite, and dolomite, but also included water-soluble components such as nitrocalcite, chloro-magnesite, and nitromagnesite. The presence of bacteria allowed for the increasing of the carbonate content by values ranging from 15% to 42%. The highest comprehensive strength was achieved for the bio-cementation solution containing urea (0.25 M), CaL (0.1 M), and an Mg/Ca molar ratio of 0.4. In the end, this research helped to achieve higher amounts of precipitates with the optimum combination of bio-cementation solutions for the soil improvement process. However, the numerical analysis of the precipitation processes and the methods reducing the environmental impact of the technology should be further investigated.

Dataset Information

Microbially induced calcite precipitation using Bacillus velezensis with guar gum.

Publications

Microbially induced calcite precipitation using Bacillus velezensis with guar gum.

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