Bioremediation of lead-contaminated mine waste by Pararhodobacter sp. based on the microbially induced calcium carbonate precipitation technique and its effects on strength of coarse and fine grained sand

Lead (Pb2+) is a toxic heavy metal that has a severe negative effect on human health and the environment. Physical, chemical and biological remediation techniques have long been used to remediate lead contamination. However, because of the great danger posed by lead contamination, there is increasing interest to apply eco-friendly and sustainable methods to remediate lead. Therefore, this study was conducted to use the microbially induced calcium carbonate precipitation (MICP) technique in conjunction with the bacterium Pararhodobacter sp. to bioremediate lead. Laboratory scale experiments were conducted and complete removal of 1036 mg/L of Pb2+ was achieved. These results were further confirmed by scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis, which indicated coprecipitation of calcium carbonate (CaCO3) and lead. The unconfined compressive strength increased with an increase in injection interval with maximum unconfined compressive strength of 1.33 MPa for fine sand, 2.87 MPa for coarse sand and 2.80 MPa for mixed sand. For Pararhodobacter sp. to efficiently induce lead immobilisation the bacterial interval required is four times with a calcium and urea concentration of 0.5 M and bacterial concentration of 109 cfu/mL. Very few low-cost in situ heavy metal treatment processes for lead bioremediation are available; therefore, bioimmobilization of lead by MICP has the potential for application as a low-cost and eco-friendly method for heavy metal remediation

Fruit and vegetable waste used as bacterial growth media for the biocementation of two geomaterials.

This paper investigates the feasibility of using randomly collected fruit and vegetable (FV) waste as a cheap growing medium of bacteria for biocementation applications. Biocementation has been proposed in the literature as an environmentally-friendly ground improvement method to increase the stability of geomaterials, prevent erosion and encapsulate waste, but currently suffers from the high costs involved, such as bacteria cultivation costs. After analysis of FV waste of varied composition in terms of sugar and protein content, diluted FV waste was used to grow ureolytic (S. pasteurii, and B.licheniformis) and also an autochthonous heterotrophic carbonic anhydase (CA)-producing B.licheniformis strain, whose growth in FV media had not been attempted before. Bacterial growth and enzymatic activity in FV were of appropriate levels, although reduced compared to commercial media. Namely, the CA-producing B.licheniformis had a maximum OD of 1.799 and a CA activity of 0.817 U/mL in FV media. For the ureolytic pathway, B. licheniformis reached a maximum OD of 0.986 and a maximum urease activity of 0.675 mM urea/min, and S. pasteurii a maximum OD  = 0.999 and a maximum urease activity of 0.756 mM urea/min. Biocementation of a clay and locomotive ash, a geomaterial specific to UK railway embankments, using precultured bacteria in FV was then proven, based on recorded unconfined compressive strengths of 1-3 MPa and calcite content increases of up to 4.02 and 8.62 % for the clay and ash respectively. Scanning Electron Microscope (SEM) and energy dispersive X-ray spectroscopy (EDS), attested the formation of bioprecipitates with characteristic morphologies and elementary composition of calcite crystals. These findings suggest the potential of employing FV to biocement these problematic geomaterials and are of wider relevance for environmental and geoenvironmental applications involving bioaugmentation. Such applications that require substrates in very large quantities can help tackle the management of the very voluminous fruit and vegetable waste produced worldwide

Use of fruit and vegetable waste as growth media in bacterial biocementation for ground improvement applications

The paper investigates the use of mixed fruit and vegetable (FV) waste to extract liquid to grow bacteria. The bacteria will be used to induce biocementation of soils and two metabolic pathways are examined. These are the ureolytic pathway and the carbonic anhydrase pathway (which absorbs CO2). The growing medium produced from fruit and vegetable waste is compared with a commercial growing medium. The results show the feasibility of using FV as a growth medium to successfully biocement soil and coal ash. A typical FV medium contains 3% total sugar and 0.302 mg/100 ml of protein. The results show that vegetable stalks and fruit peel media support the growth of both ureolytic bacteria B. licheniformis and U-1, a carbonic anhydrase-producing bacteria. The use of FV waste to grow bacteria leads to a reduction in biocementation costs for ground improvement applications

Concurrent Carbon Capture and Biocementation through the Carbonic Anhydrase (CA) Activity of Microorganisms -a Review and Outlook, Environmental Processes

Biocementation, i.e., the production of biomimetic cement through the metabolic activity of microorganisms, offers exciting new prospects for various civil and environmental engineering applications. This paper presents a systematic literature review on a biocementation pathway, which uses the carbonic anhydrase (CA) activity of microorganisms that sequester CO2 to produce biocement. The aim is the future development of this technique for civil and (geo-)environmental engineering applications towards CO2-neutral or negative processes. After screening 248 potentially relevant peer-reviewed journal papers published between 2002 and 2023, 38 publications studying CA-biocementation were considered in the review. Some of these studies used pure CA enzyme rather than bacteria-produced CA. Of these studies, 7 used biocementation for self-healing concrete, 6 for CO2 sequestration, 10 for geotechnical applications, and 15 for (geo-)environmental applications. A total of 34 bacterial strains were studied, and optimal conditions for their growth and enzymatic activity were identified. The review concluded that the topic is little researched; more studies are required both in the laboratory and field (particularly long-term field experiments, which are totally lacking). No studies on the numerical modelling of CA-biocementation and the required kinetic parameters were found. The paper thus consulted the more widely researched field of CO2 sequestration using the CA-pathway, to identify other microorganisms recommended for further research and reaction kinetic parameters for numerical modelling. Finally, challenges to be addressed and future research needs were discussed

Synthesis, characterisation, and utilisation of copper nanoflower for biocementation for ground improvement applications

Microbially-induced calcium carbonate precipitation (MICP) has recently emerged as a sustainable ground improvement method. Nevertheless, the technique’s applicability in soils with narrow pore throats has been queried. To overcome these challenges, the use of enzymes (including bacterially produced enzymes) was proposed for these soils. However, the use of free enzymes entails many challenges linked predominantly to the limited enzyme supply, the poor stability of the enzyme once released into the soil, and the poor reusability of the enzyme. This paper studies the use of nano enzymes with a high biocementation efficacy for carbonic anhydrase (CA) enzyme delivery as one possible way to overcome potentially these challenges. CA enzyme was used because it has the potential to be an environmentally sustainable biocementation pathway due to its ability to sequester CO2 for biocement production. The paper presents the synthesis, characterisation, and utilisation of CA-enwrapped copper phosphate-based inorganic hybrid nanoflowers for innovative delivery and enzyme stabilisation due to the enhanced thermal and enzyme activity efficiency and due to their reusability, if recovered at the end of the treatment. The results from this study show that the bovine carbonic anhydrase enzyme enhanced the CO2 hydration reaction, resulting in a bioprecipitation reaction and the production of calcium carbonate and increased strength of treated soil with 500kPa for free CA and approximately 1000kPa for the hybrid CA-Cu. The material analysis confirmed calcite as the primary precipitate formed, which would act as a bonding agent between soil particles for ground improvement applications

Synthesis and Utilisation of Hybrid Metal-Carbonic Anhydrase Enzyme Carrier System for Soil Biocementation

Biocementation is an emerging nature-inspired method of producing eco-friendly cement for soil stabilization. This paper used the bovine-derived carbonic anhydrase (CA) enzyme to catalyse the bioprecipitation of CaCO3 in a fine-grained soil and thus to biocement the soil. To increase the efficiency of the CA, an innovative copper–carbonic anhydrase (CA) hybrid was fabricated. This study is a proof-of-concept of the potential application of these enzyme carriers for soil bioce-mentation. The hybrid carriers are aimed to enhance the stability, recovery and reusability of the enzyme used in the biocementation process. The results showed that the fabricated copper phosphate-based inorganic hybrid was stable throughout the duration of the tests (2 months) and under a wide range of pH and temperatures. Its enzymatic activity was enhanced compared to the free CA enzyme and it was proved suitable for soil biocementation. This was further confirmed by the SEM analysis. Additionally, the treated soil with the formulated hybrid carrier showed im-proved unconfined compressive strength, especially when the carriers were implemented into the soil by mixing. The material analysis by Raman spectroscopy confirmed calcium carbonate as the primary precipitate, consistent with soil biocementation. Overall, this innovative method of de-livery of enzymes with enhanced stability and activity shows promise that, upon further devel-opment, it can be successfully used to increase the efficiency and sustainability of the biocemen-tation process

Biocementation mediated by native Carbonic Anhydrase-producing microbes.

This study investigated the feasibility of biocementing a fine-grained foundation soil from the East Anglia railway network via the carbonic anhydrase (CA) pathway. This pathway is a promising way of improving the mechanical properties of soils by biocementation while sequestering CO2 during the process. To achieve the aim of this research, forty CA-producing bacterial isolates from soil layers below a railway embankment in East Anglia, UK, were screened and selected using a qualitative CA activity assay. Three of these bacteria expressed high and stable CA enzyme activity and were further characterised by their morphological, molecular, and enzyme profile characteristics. Bioaugmentation was then employed to biocement the soil from the site using the native CA-producing bacteria isolated from the soil. The unconfined compressive strength and calcite content of the treated soil were determined. Preliminary results showed a substantial increase in soil unconfined compressive strength upon biocementation treatment. Although further geotechnical testing is the subject of future work, the unconfined compressive strength and calcite content results obtained so far proved biocementation of the fine-grained soil and showed promise that the CA biocementation route can be further developed as a successful and environmentally friendly soil stabilization technique, with the added advantage of sequestering CO2 from the atmosphere or using captured waste CO2, during the biocementation process

Mechanism of salinity change and hydrogeochemical evolution of groundwater in the Machile-Zambezi Basin, South-western Zambia

Machile-Zambezi Basin, South-Western Zambia hosts high salinity groundwater which threatens water security for rural inhabitants. This study investigates the hydrological mechanism that led to high salinity and the geochemical evolution of the groundwater system. The Machile-Zambezi Basin is part of the wider Kalahari Basin which underwent major palaeo-environmental climatic, tectonic and sedimentology dynamics which must have impacted the groundwater salinity. The study examines the groundwater level, hydrochemistry, environmental isotopes (18O/16O, 2H/1H, 3H/3He, 14C/13C). In addition, the sediment cation exchange capacity (CEC) and pore-water chemistry on intact core material were measured. The groundwater chemistry evolved from fresh Ca(Na)HCO3 to saline Na(Ca, Mg)SO4 due to dissolution of salts and not evaporation as indicated by stable isotopes. The saline groundwater is old with 14C ages estimates of more than 1000 years old and stagnant. Geochemical modelling using PHREEQC suggests that ionic exchange due to release of cations from dissolving salts and sulphate reduction were also important processes in this system. High groundwater salinity is therefore associated with Pre-Holocene environmental changes and is restricted to a stagnant saline zone. It will therefore remain unflushed as long as current climatic conditions remain

Concurrent Carbon Capture and Biocementation through the Carbonic Anhydrase (CA) activity of microorganisms ‑ a review and outlook

Biocementation, i.e., the production of biomimetic cement through the metabolic activity of microorganisms, offers exciting new prospects for various civil and environmental engineering applications. This paper presents a systematic literature review on a biocementation pathway, which uses the carbonic anhydrase (CA) activity of microorganisms that sequester CO2 to produce biocement. The aim is the future development of this technique for civil and (geo-)environmental engineering applications towards CO2-neutral or negative processes. After screening 248 potentially relevant peer-reviewed journal papers published between 2002 and 2023, 38 publications studying CA-biocementation were considered in the review. Some of these studies used pure CA enzyme rather than bacteria-produced CA. Of these studies, 7 used biocementation for self-healing concrete, 6 for CO2 sequestration, 10 for geotechnical applications, and 15 for (geo-)environmental applications. A total of 34 bacterial strains were studied, and optimal conditions for their growth and enzymatic activity were identified. The review concluded that the topic is little researched; more studies are required both in the laboratory and field (particularly long-term field experiments, which are totally lacking). No studies on the numerical modelling of CA-biocementation and the required kinetic parameters were found. The paper thus consulted the more widely researched field of CO2 sequestration using the CA-pathway, to identify other microorganisms recommended for further research and reaction kinetic parameters for numerical modelling. Finally, challenges to be addressed and future research needs were discussed