• Proceedings of the Korea Concrete Institute Conference
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• 2009.05a
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• pp.525-526
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• 2009

On this paper we induce calcite($CaCO_3$) precipitation using microbial biomineralization of the Sporosarcina pasteurii and evaluate required performance evaluation by adjusting it to mortar. As a result carbonation normal mortar test piece(C3S-W) and mortar test piece(C3S-S.p) mixed with Sporosarcina pasteurii, reaction of C3S-S.p was late than C3S-W. Also, in the case of carbonation experiment of C3S-S.p curing in the Urea-CaCl2 aqueous solution(Medium) during 28days and durability of the C3S-W, durability of the mortar test piece(C3S-S.p) mixed with Sporosarcina pasteurii become higher than normal mortar test piece(C3S-W).

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.540-548
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• 2016

Microbially induced calcium carbonate precipitation (CCP) is a long-standing but re-emerging environmental engineering process for production of self-healing concrete, bioremediation, and long-term storage of CO₂. CCP-capable bacteria, two Bacillus strains (JH3 and JH7) and one Sporosarcina strain (HYO08), were isolated from two samples of concrete and characterized phylogenetically. Calcium carbonate crystals precipitated by the three strains were morphologically distinct according to field emission scanning electron microscopy. Energy dispersive X-ray spectrometry mapping confirmed biomineralization via extracellular calcium carbonate production. The three strains differed in their physiological characteristics: growth at alkali pH and high NaCl concentrations, and urease activity. Sporosarcina sp. HYO08 and Bacillus sp. JH7 were more alkali- and halotolerant, respectively. Analysis of the community from the same concrete samples using barcoded pyrosequencing revealed that the relative abundance of Bacillus and Sporosarcina species was low, which indicated low culturability of other dominant bacteria. This study suggests that calcium carbonate crystals with different properties can be produced by various CCP-capable strains, and other novel isolates await discovery.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1328-1338
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• 2015

Years of research have shown that the application of microorganisms increases the compressive strength of cement-based material when it is cured in a culture medium. Because the compressive strength is strongly affected by the hydration of cement paste, this research aimed to investigate the role of the microorganism Sporosarcina pasteurii in hydration of cement paste. The microorganism's role was investigated with and without the presence of a urea-CaCl₂ culture medium (i.e., without curing the specimens in the culture medium). The results showed that S. pasteurii accelerated the early hydration of cement paste. The addition of the urea-CaCl₂ culture medium also increased the speed of hydration. However, no clear evidence of microbially induced calcite precipitation appeared when the microorganisms were directly mixed with cement paste.

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.312-319
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• 2002

The Sporosarcina psychrophilia pyrB gene, which encodes aspartate transcarbamylase (ATcase), was cloned on Sau3AI restriction endonuclease fragment inserted into pUC19 plasmid vector, S. psychrophilia pyrB gene was expressed in Escherichia coli pyrB mutant for the complementation test. The sequence of 2,606 nucleotides including putative pyrB gene was determined. The region contained one full open reading frame (ORf) and two partial ORFs. The deduced amino acid sequence of the second ORF showed 59% identity with that of Bacillus caldolyticus ATCase. The first and third partial ORFs were closely related to the uracil permease (pyrP) and dihydroorotase (pyrC), respectively. Besides, potential terminator, antiterminator, and anti-antiterminator structures were found in the intergenic region between pyrP and pyrB. These results suggested that S. psychrophilia pyrimidine nucleotide biosynthesis genes are clustered as well as other Bacillus sp. Over-expressed product of pyrB encoding ATCase was purified and analyzed by the SDS-PAGE. The purified PyrB protein turned out to be molecular mass of 27 kDa and showed ATCase activity.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.255-259
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• 1999

A 6.9-kb DNA fragment including the minimal Bacillus pasteurii urease gene cluster was subcloned into a high-copy-number plasmid vector, pUC19, and the recombinant B. pasteurii urease was overexpressed in Escherichia coli. The recombinant urease was purified 25.9-fold by using combinations of anion-exchange and gel-filtration chromatography followed by Mono-Q chromatography on a FPLC. N-terminal peptide sequencing analyses revealed that two distinct smaller peptide bands resolved on a 10-18％ gradient SDS-PAGE corresponded to UreA and UreB peptides, respectively. It was also shown that the ureB gene was translated from a GUG codon and the first methionine residue was post-translationally cleaved off. The native molecular weight of the recombinant urease was 176,000 and 2 nickel atoms were present per catalytic unit. pH stability studies of the purified enzyme showed that the recombinant Bacillus pasteurii urease is stable in alkaline pH range, which is similar to the enzyme of the evolutionarily related bacterium, Sporosarcina ureae.

• Journal of the Korea Concrete Institute
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• v.22 no.4
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• pp.547-557
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• 2010

This paper presents a study on the development of next generation smart concrete in an eco-friendly manner using micro-biologically induced calcite precipitation (MICP) via microbial biomineralization. It seems that currently, the reformation and functional improvement of concrete using MICP can be achieved using Sporosarcina pasteurii, which is a representative microorganism that produces calcite precipitation. Based on previous studies on MICP the biochemical tests and crystallinity evaluation of cement using sporoasrcina pasteurii and four additional micro-organisms from the concrete structures as identified by 16S rDNA sequence analysis were conducted. Also by applying the Sporosarcina pasteurii and separated four effective micro-organisms from the concrete structures to mortar, the compressive strength improvement by varying curing conditions, repair of crack were examined, and plans for future study were suggested. The effect of the application of effective micro-organisms can lead to the development of a new material that will contribute to resolution of environmental problems and facilitate repair work, and this can also serve as a new research theme in the future. In addition, the importance of this study is to use micro-organism, which is found common in concrete structures, this new microbial is not only environmentally safe but also persists in the natural environment for an extended period of time. Therefore, it seems to have a great potential to became a new environmentally low-burdened functional material.

• Journal of Soil and Groundwater Environment
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• v.20 no.3
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• pp.83-91
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• 2015

Mine soil contamination by high levels of metal ions that prevents the successful vegetation poses a serious problem. In the study presented here, we used the microbial biocatalyst of urease producing bacterium Sporosarcina pasteurii or plant extract based BioNeutro-GEM (BNG) agent. The ability of the biocatalysts to bioremediate contaminated soil from abandoned mine was examined by solid-state composting vegetation under field conditions. Treatment of mine soil with the 2 biocatalysts for 5 months resulted in pH increase and electric conductivity reduction compared to untreated control. Further analyses revealed that the microbial catalysts also promoted the root and shoot growth to the untreated control during the vegetation treatments. After the Sporosarcina pasteurii or plant extract based BNG treatment, the microbial community change was monitored by culture-independent pyrosequencing. These results demonstrate that the microbial biocatalysts could potentially be used in the soil bioremediation from mine-impacted area.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.184.3-184
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• 2005

• Journal of the Korean Geotechnical Society
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• v.28 no.3
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• pp.67-75
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• 2012

This paper presents an environment-friendly sand cementation method by precipitating calcium carbonate using plant extracts. The plant extracts contain urease like $Sporosarcina$ $pasteurii$, which can decompose urea into carbonate ion and ammonium ion. It can cause cementation within sand particles where carbonate ions decomposed from urea combine with calcium ions dissolved from calcium chloride or calcium hydroxide to form calcium carbonate. Plant extracts, urea and calcium chloride or calcium hydroxide were blended and then mixed with Nakdong River sand. The mixed sand was compacted into a cylindrical specimen and cured for 3 days at room temperature ($18^{\circ}C$). Unconfined compression test, SEM and XRD analyses were carried out to evaluate three levels of urea concentration and two different calcium sources. As urea concentration increased, the unconfined compressive strength increased up to 10 times those without plant extracts because calcium carbonate precipitated more, regardless of calcium source. It was also found that the strength of specimen using calcium chloride was higher than that of specimen using calcium hydroxide.

• Korean Journal of Soil Science and Fertilizer
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• v.47 no.6
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• pp.391-397
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• 2014

Acid mine drainage occurrence is a serious environmental problem by mining industry; it usually contain high levels of metal ions, such as iron, copper, zinc, aluminum, and manganese, as well as metalloids of which arsenic is generally of greatest concern. It causes mine impacted soil pollution with mining and smelting activities, fossil fuel combustion, and waste disposal. In the present study, three bacterial strains capable of producing urease were isolated by selective enrichment of heavy metal contaminated soils from a minei-mpacted area. All isolated bacterial strains were identified Sporosarcina pasteurii with more than 98% of similarity, therefore they were named Sporosarcina sp. KM-01, KM-07, and KM-12. The heavy metals detected from the collected mine soils containing bacterial isolates as Mn ($170.50mg\;kg^{-1}$), As ($114.05mg\;kg^{-1}$), Zn ($92.07mg\;kg^{-1}$), Cu ($62.44mg\;kg^{-1}$), and Pb ($40.29mg\;kg^{-1}$). The KM-01, KM-07, and KM-12 strains were shown to be able to precipitate calcium carbonate using urea as a energy source that was amended with calcium chloride. SEM-EDS analyses showed that calcium carbonate was successfully produced and increased with time. To confirm the calcium carbonate precipitation ability, urease activity and precipitate weight were also measured and compared. These results demonstrate that all isolated bacterial strains could potentially be used in the bioremediation of acidic soil contaminated by heavy metals by mining activity.