rexresearch

Microbially Induced Calcite Precipitation / MICP
( Ureolytic Calcite Precipitation / UCP )

Relate : HILBERTZ: BioRock .. DAVIDOVITS: GeoPolymers ... HWANG: Grancrete ... ROTTA LORIA: ElectroCement ... Hydrophobic Cement ...


https://pmc.ncbi.nlm.nih.gov/articles/PMC10467343/
Appl Environ Microbiol. 2023 Jul 13;89(8):e01794-22. doi: 10.1128/aem.01794-22

Microbially Induced Calcium Carbonate Precipitation by Sporosarcina pasteurii: a Case Study in Optimizing Biological CaCO3 Precipitation
Michael S Carter, et al.
[ PDF ]

ABSTRACT -- Current production of traditional concrete requires enormous energy investment that accounts for approximately 5 to 8% of the world’s annual CO2 production. Biocement is a building material that is already in industrial use and has the potential to rival traditional concrete as a more convenient and more environmentally friendly alternative. Biocement relies on biological structures (enzymes, cells, and/or cellular superstructures) to mineralize and bind particles in aggregate materials (e.g., sand and soil particles). Sporosarcina pasteurii is a workhorse organism for biocementation, but most research to date has focused on S. pasteurii as a building material rather than a biological system. In this review, we synthesize available materials science, microbiology, biochemistry, and cell biology evidence regarding biological CaCO3 precipitation and the role of microbes in microbially induced calcium carbonate precipitation (MICP) with a focus on S. pasteurii. Based on the available information, we provide a model that describes the molecular and cellular processes involved in converting feedstock material (urea and Ca2+) into cement. The model provides a foundational framework that we use to highlight particular targets for researchers as they proceed into optimizing the biology of MICP for biocement production.



US11512021 -- PREVENTING OR REDUCING PLANT GROWTH BY BIOCEMENTATION  

...Mixture 4 was composed of the following components in the following concentrations:

20.0  g/l  Yeast extract
0.25  M  calcium chloride
18.0  g/l  urea
4 × 10<{circumflex over ( )}>8  cells/ml  Sp. pasteurii

The mixture also contained trace elements and traces of salts and sugars, for example (<1%). In this medium, urea served primarily as a source of carbonate and secondarily as a source of nitrogen.

In mixture 5, 50 ml/l Silicade 8 (silica sol-acrylic dispersion) was additionally added as additive. The additive was used to achieve a longer lasting stability of the biocementation layer.

The components of the biocementation mixtures 4 and 5 (without bacteria) were present in solid form, respectively. The bacteria were present as liquid culture in a culture medium known from the state of the art, respectively, as described for example in Cuthbert, M. O. et al., Ecological Engineering 2012, 41, 32-40 (see section 2.2, p. 33). The solid components and the bacteria in liquid culture were mixed directly before use, respectively, dissolving the solid components....



http://dustbiosolutions.com/
Dust BioSolutions GmbH
Am Klopferspitz 19
82152 Planegg
Germany



https://bioplasticsnews.com/2017/06/12/turning-dust-into-stone-with-bacteria-martin-spitznagel-founder-dust-biosolutions/
Turning Dust Into Stone with Bacteria by Martin Spitznagel founder of Dust Biosolutions GmbH

The company has developed a portfolio of 30 liquid biological binders able to work with sand stone, dusts, etc. The technology is called bio-cementation and use calcite producing bacteria. It takes 24 to 48h to be in full effect and support e.g. the weight of a heavy truck. It is very powerful to encapsulate dusts associated with extractive industries and is reportedly 34% cheaper than the cheapest chemical spray. It does not need to be used more than once or twice a year contrarily to water that requires spraying several times a day. It is not harmful to the environment and biodegrades. Next to the recent financing round, the company name will change from the current project name. Biology PhDs are in urgent need...


Some MICP Patents:

CN121571451 -- Soil remediation method and system based on EKG electric power and MICP
Abstract -- The invention belongs to the technical field of soil remediation. According to the soil remediation method and system based on the EKG electric power and the MICP, a horizontal landfill type implementation mode and a vertical insertion type implementation mode are adopted, conductive plastic electrodes (including an anode and a cathode) are laid, and a drip irrigation system, a high-pressure liquid injection system, a vacuum negative pressure pumping drainage system and a soil moisture content, pollutant and pH value monitoring instrument are matched; firstly, water-soluble and free-state pollutants are directionally migrated, pumped and removed by utilizing an electric field effect, microorganisms such as urease bacteria are injected through a high-pressure liquid injection pipeline in the later period of restoration, and residual heavy metal is converted into carbonate combined-state precipitates by virtue of an MICP technology to be stable. According to the invention, dual remediation of'removal-stabilization 'is realized, secondary pollution is avoided, the intelligent degree is high, the system is suitable for river, lake and reservoir dredged sediment, coastal mud flat dredger fill, petrochemical engineering sites and other scenes, and the content of soil pollutants is ensured to reach the environmental quality standard or design standard

CN121573935 -- Solid waste landfill crack combination self-repairing material based on microbial mineralization and application
Abstract -- The invention discloses a solid waste landfill crack combination self-repairing material based on microbial mineralization and application, and belongs to the field of environmental geotechnical engineering. The method comprises the steps that firstly, a klebsiella liquid and a calcium source cementing liquid are prepared, grouting holes are formed in the side slope, the bacterial liquid and the cementing liquid are injected in batches, and internal cracks are filled with a microorganism induced calcium carbonate precipitation (MICP) technology; and then, a fiber reinforced guniting material formed by compounding field soil, active magnesium oxide and chopped fibers is adopted to spray and cover the surface layer of the slope, and a compact sealing layer is formed through carbon dioxide carbonization. By combining internal MICP curing and surface layer fiber reinforced magnesium oxide carbonization sealing, a synergistic protection system with strong inside and tough outside is constructed, the anti-permeability and stability of the slope are improved, meanwhile, the toughness, crack resistance and long-term durability of a surface layer sealing layer are remarkably enhanced, and the service life of the slope is prolonged. And the method is particularly suitable for slope treatment of projects with high anti-seepage requirements such as solid waste landfills and the like

CN121451579 -- Reinforcing method for MICP-bionic structure interlayer of coral sand soil body
Abstract -- The invention relates to the technical field of buildings, in particular to a coral sand soil body MICP-bionic structure interlayer reinforcing method which comprises the following steps: S1, grading original coral sand, taking fine particles as an interlayer material, and taking medium-coarse particles as a main body filling material; s2, laying a 3D printing bionic geosynthetic material in the fine particles, sequentially injecting a sporosarcina pasteurii bacterial solution and a urea-calcium chloride cementing solution, standing, and alternately grouting for multiple times; s3, the composite interlayer and the main body sand layer are stacked and compacted in a layered mode, and a layered composite structure is constructed; and S4, repeating the steps S1 to S3. The ecological reinforcement technology and the reinforcement technology are combined, a collaborative system of layered construction, interlayer construction and overall reinforcement is constructed, and the reinforcement effect and efficiency are effectively improved.

CN121426590 -- Carbonate rock cultural relic restoration method based on MICP
Abstract -- The invention discloses a carbonate rock cultural relic restoration method based on MICP, and belongs to the technical field of cultural relic restoration. A carbonate rock cultural relic restoration method based on MICP comprises the following steps that S1, early-stage detection is conducted, specifically, mineral composition, porosity and microfracture distribution are conducted on carbonate rock cultural relics to be restored, the surface weathering degree and spatial distribution characteristics are evaluated, the pH, the water content and the ion composition of a restoration area are measured, and restoration parameters are determined; s2, bacterial liquid preparation and activity control; s3, repairing application: applying the bacterial liquid and the calcium source solution to the target area in a spraying or permeating manner according to the shape and damage degree of the cultural relics; s4, curing and curing; and S5, environment restoration and in-situ control. Calcium carbonate is induced by microorganisms to be deposited on the surface and in cracks of the carbonate rock cultural relic to form newly generated calcium carbonate filler, and the overall stability and weather resistance of the cultural relic structure are improved.

CN121407588 -- Slope grouting reinforcement device and method based on MICP technology
Abstract -- The invention relates to the technical field of road slope grouting, in particular to a slope grouting reinforcement device and method based on the MICP technology. The device comprises a bearing vehicle, a rotating disc, a first telescopic arm, a second telescopic arm, a liquid storage tank, a pump machine, a pipe winding and unwinding disc and a conveying pipe. The turntable is rotationally connected with the bearing vehicle; the other end of the first telescopic arm is fixedly connected with the second telescopic arm; the second telescopic arm is perpendicular to the first telescopic arm; the free end of the second telescopic arm is detachably connected with a spray head; the liquid storage tank, the pump machine and the winding and unwinding pipe disc are all fixedly mounted on the bearing vehicle, and the conveying pipe is wound on the winding and unwinding pipe disc; a liquid inlet of the pump machine is connected to the liquid storage tank, a liquid outlet is connected to the liquid inlet end of the conveying pipe, and the liquid outlet end of the conveying pipe is connected to the spray head. The device can efficiently conduct slope grouting operation on the whole road slope; and the distance between the spray head and the road slope can be conveniently adjusted, and it can be guaranteed that the reinforcing depth and the compaction degree of the whole slope tend to be consistent

CN121275450 -- Sandy soil solidification method based on synergistic effect of MICP and organic polymer emulsion
Abstract -- The invention provides a sand solidification method based on the synergistic effect of MICP and organic polymer emulsion, which is characterized by comprising the following steps: filling sand into a mold, oscillating and compacting to manufacture a sand column; activating the microbial powder with the mineralization effect to obtain a bacterial liquid; mixing urea and calcium chloride to obtain a cementing solution, and then adding the waterborne acrylic epoxy hybrid emulsion to obtain a mixed solution; pouring the bacterial liquid into the sand column until the bacterial liquid is completely immersed, and standing until bacteria in the bacterial liquid are fully attached to the sand particles; filling the mixed liquid into a sand column to finish a round of biological grouting; and carrying out multiple rounds of biological grouting until the sand column cannot be filled, and after the mineralization reaction is completed, demolding, cleaning and drying the sand column to obtain the cured sand column. The mechanical property of a sand body is greatly improved by forming an organic film-calcium carbonate inorganic framework synergistic cementation structure, particularly through a treatment group of multiple rounds of mixed grouting, the unconfined compressive strength is improved more remarkably compared with that of a traditional MICP technology, and the requirement of heavy-load engineering for the foundation strength can be met.



https://en.wikipedia.org/wiki/Sporosarcina_pasteurii
Sporosarcina pasteurii

Sporosarcina pasteurii formerly known as Bacillus pasteurii from older taxonomies, is a gram positive bacterium with the ability to precipitate calcite and solidify sand given a calcium source and urea; through the process of microbiologically induced calcite precipitation (MICP) or biological cementation.[2] S. pasteurii has been proposed to be used as an ecologically sound biological construction material. Researchers studied the bacteria in conjunction with plastic and hard mineral; forming a material stronger than bone.[3] It is a commonly used for MICP since it is non-pathogenic and is able to produce high amounts of the enzyme urease which hydrolyzes urea to carbonate and ammonia.[4]
Physiology

S. pasteurii is a gram positive bacterium that is rod-like shaped in nature. It has the ability to form endospores in the right environmental conditions to enhance its survival, which is a characteristic of its bacillus class.[5] It has dimensions of 0.5 to 1.2 microns in width and 1.3 to 4.0 microns in length. Because it is an alkaliphile, it thrives in basic environments of pH 9–10. It can survive relatively harsh conditions up to a pH of 11.2.[4]
Metabolism and growth

S. pasteurii are soil-borne facultative anaerobes that are heterotrophic and require urea and ammonium for growth.[6] The ammonium is utilized in order to allow substrates to cross the cell membrane into the cell.[6] The urea is used as the nitrogen and carbon source for the bacterium. S. pasteurii are able to induce the hydrolysis of urea and use it as a source of energy by producing and secreting the urease enzyme. The enzyme hydrolyzes the urea to form carbonate and ammonia. During this hydrolysis, a few more spontaneous reactions are performed. Carbamate is hydrolyzed to carbonic acid and ammonia and then further hydrolyzed to ammonium and bicarbonate.[4] This process causes the pH of the reaction to increase 1–2 pH, making the environment more basic which promotes the conditions that this specific bacterium thrives in.[7] Maintaining a medium with this pH can be expensive for large scale production of this bacterium for biocementation. A wide range of factors can affect the growth rate of S. pasteurii. This includes finding the optimal temperature, pH, urea concentration, bacterial density, oxygen levels, etc.[7] It has been found that the optimal growing temperature is 30 °C, but this is independent of the other environmental factors present.[5] Since S. pasteurii are halotolerant, they can grow in the presence of low concentrations of aqueous chloride ions that are low enough to not inhibit bacterial cell growth.[7] This shows promising applications for MICP use.

S. pasteurii DSM 33 is described to be auxotrophic for L-methionine, L-cystein, thiamine and nicotinic acid.[8]

Genomic properties

The whole genome of S. pasteurii NCTC4822 was sequenced and reported under NCBI Accession Number: NZ_UGYZ01000000. With a chromosome length of 3.3 Mb, it contains 3,036 protein coding genes and has GC content of 39.17% .[9] When the ratio of known functional genes to the unknown genes is calculated, the bacterium shows highest ratios for transport, metabolism, and transcription. The high proportion of these functions allows the conversion of urea to carbonate ions which is necessary for the bio-mineralization process.[9] The bacterium has seven identified genes that are directly related to urease activity and assembly as well, which can be further studied to give insight about maximizing urease production for optimizing use of S. pasteurii in industrial applications.[9]

Applications with MICP

S. pasteurii have the unique capability of hydrolyzing urea and through a series of reactions, produce carbonate ions. This is done by secreting copious amounts of urease through the cell membrane.[5] When the bacterium is placed in a calcite rich environment, the negatively charged carbonate ions react with the positive metal ions like calcium to precipitate calcium carbonate, or bio-cement.[4] The calcium carbonate can then be used as a precipitate or can be crystallized as calcite to cement sand particles together. Therefore, when put into a calcium chloride environment, S. pasteurii are able to survive since they are halotolerant and alkaliphiles. Since the bacteria remain intact during harsh mineralization conditions, are robust, and carry a negative surface charge, they serve as good nucleation sites for MICP.[9] The negatively charged cell wall of the bacterium provides a site of interaction for the positively charged cations to form minerals. The extent of this interaction depends on a variety of factors including the characteristics of the cell surface, amount of peptidoglycan, amidation level of free carboxyl, and availability of teichoic acids.[7] S. pasteurii show a highly negative surface charge which can be shown in its highly negative zeta potential of −67 mV compared to non-mineralizing bacteria E. coli, S. aureus and B. subtilis at −28, −26 and −40.8 mV, respectively.[9] Aside from all of these benefits towards using S. pasteurii for MICP, there are limitations like undeveloped engineering scale-up, undesired by-products, uncontrolled growth, or dependence on growth conditions like urea or oxygen concentrations.[9]

Current and potential applications

S. pasteurii have a purpose in improving construction material as in concrete or mortar. Concrete is one of the most used materials in the world but it is susceptible to forming cracks which can be costly to fix. One solution is to embed this bacterium in the cracks and once it is activated using MICP. Minerals will form and repair the gap in a permanent environmentally-friendly way. One disadvantage is that this technique is possible only for external surfaces that are reachable.[7]

Another application is to use S. pasteurii in bio self-healing of concrete which involves implementing the bacterium into the concrete matrix during the concrete preparation to heal micro cracks. This has a benefit of minimal human intervention and yields more durable concrete with higher compressive strength.[7]

One limitation of using this bacterium for bio-mineralization is that although it is a facultative anaerobe, in the absence of oxygen, the bacterium is unable to synthesize urease anaerobically. A lack of oxygen also prevents MICP since its initiation relies heavily on oxygen. Therefore, at sites distant from the injection location or at great depths, the likelihood of precipitation decreases.[9] One potential fix is to couple this bacterium in the biocement with oxygen releasing compounds (ORCs) that are typically used for bioremediation and removal of pollutants from soil.[7] With this combination, the lack of oxygen can be diminished and the MICP can be optimized with the bacterium.

Some specific examples of current applications include:

Architecture student Magnus Larsson won the 2008 Holcim Award "Next Generation" first prize for region Africa Middle East for his project "Dune anti-desertification architecture, Sokoto, Nigeria" and his design of a habitable wall.[10] Larssons also presented the proposal at TED.[11]

Ginger Krieg Dosier's unique biotechnology start-up company, bioMason, in Raleigh, NC has developed a method of growing bricks from Sporosarcina pasteurii and naturally abundant materials. In 2013 this company won the Cradle to Cradle Innovation Challenge (which included a prize of $125,000) and the Dutch Postcode Lottery Green Challenge (which included a prize of 500,000 euros).[12]

More potential applications include:

Use bacteria to solidify liquefiable soils in areas prone to earthquakes.
Form bio-bricks
Stabilize marshes and swamps
Reduce the settlement rate of buildings[6]
Remove heavy metals from wastewater[13]
used as barrier for weed control in agriculture, as an alternative to herbicide[14]

Considerations of using this bacterium in industrial applications is scale-up potential, economic feasibility, long-term viability of bacteria, adhesion behavior of calcium carbonate, and polymorphism.[7]

References

 "Species: Sporosarcina pasteurii". lpsn.dsmz.de. Archived from the original on 17 June 2024. Retrieved 17 June 2024.
   
 Chou CW, Aydilek A, Seagren E, Maugel T (November 2008). "Bacterially-induced calcite precipitation via ureolysis". American Society for Microbiology.

    "Microbial makers help humans to build tough stuff". Nature. 591 (7849): 180. 4 March 2021. Bibcode:2021Natur.591R.180.. doi:10.1038/d41586-021-00565-3.

    Henze J, Randall DG (August 2018). "Microbial induced calcium carbonate precipitation at elevated pH values (>11) using Sporosarcina pasteurii". Journal of Environmental Chemical Engineering. 6 (4): 5008–5013. doi:10.1016/j.jece.2018.07.046. S2CID 105388152.

    Bhaduri S, Debnath N, Mitra S, Liu Y, Kumar A (April 2016). "Microbiologically Induced Calcite Precipitation Mediated by Sporosarcina pasteurii". Journal of Visualized Experiments (110). doi:10.3791/53253. PMC 4941918. PMID 27167458.

    "Optimizing the use of sporosarcina pasteurii bacteria for the stiffening of sand". www.envirobiotechjournals.com. Archived from the original on 17 June 2024. Retrieved 4 May 2020.

    Seifan M, Berenjian A (November 2018). "Application of microbially induced calcium carbonate precipitation in designing bio self-healing concrete". World Journal of Microbiology & Biotechnology. 34 (11): 168. doi:10.1007/s11274-018-2552-2. PMID 30387067. S2CID 53295171.
   
 Lapierre FM, Schmid S, Ederer B, Ihling N, Büchs J, Huber R (December 2020). "Revealing nutritional requirements of MICP-relevant Sporosarcina pasteurii DSM33 for growth improvement in chemically defined and complex media". Scientific Reports. 10 (22448): 22448. Bibcode:2020NatSR..1022448L. doi:10.1038/s41598-020-79904-9. PMC 7775470. PMID 33384450.
   
 Ma L, Pang AP, Luo Y, Lu X, Lin F (January 2020). "Beneficial factors for biomineralization by ureolytic bacterium Sporosarcina pasteurii". Microbial Cell Factories. 19 (1): 12. doi:10.1186/s12934-020-1281-z. PMC 6979283. PMID 31973723.
   
 Holcim Awards 2008 Africa Middle East "Next Generation" 1st prize: Dune anti-desertification architecture, Sokoto, Nigeria, Holcim awards. Retrieved 20 February 2010.
   
 Magnus Larsson: Dune architect , TED.com. Retrieved 20 February 2010.
    bioMason @Green Challenge
    
Torres-Aravena, Álvaro Esteban; Duarte-Nass, Carla; Azócar, Laura; Mella-Herrera, Rodrigo; Rivas, Mariella; Jeison, David (November 2018). "Can Microbially Induced Calcite Precipitation (MICP) through a Ureolytic Pathway Be Successfully Applied for Removing Heavy Metals from Wastewaters?". Crystals. 8 (11): 438. doi:10.3390/cryst8110438.