• KSBB Journal
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• v.29 no.5
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• pp.323-327
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• 2014

Microbially induced calcite precipitation is a naturally occurring biological process in which microbes produce calcite on the surface of the microorganisms by urease activity. In order to collect calcite forming bacteria (CFB) in Korea, we isolated 343 putative CFB strains from various environments over three year period (2011~2013) and selected 100 CFB strains. Average of calcite productivity was 10.56 mg/mL. And average of ammonium concentration by urease activity was $8.00{\mu}M$. Two useful CFB strains of the others were analyzed by 16S rRNA and identified as Sporosarcina sp. and Viridibacillus arenosi. The CFB strains presented in this study are indigenous microorganisms in Korea and they are expected to be applicable to a variety of environments in the country.

• Journal of Microbiology and Biotechnology
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• v.23 no.9
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• pp.1269-1278
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• 2013

Crack remediation on the surface of cement mortar using microbiological calcium carbonate ($CaCO_3$) precipitation (MICP) has been investigated as a microbial sealing agent on construction materials. However, MICP research has never acknowledged the antifungal properties of calcite-forming bacteria (CFB). Since fungal colonization on concrete surfaces can trigger biodeterioration processes, fungi on concrete buildings have to be prevented. Therefore, to develop a microbial sealing agent that has antifungal properties to remediate cement cracks without deteriorative fungal colonization, we introduced an antifungal CFB isolated from oceanic islands (Dokdo islands, territory of South Korea, located at the edge of the East Sea in Korea.). The isolation of CFB was done using B4 or urea-$CaCl_2$ media. Furthermore, antifungal assays were done using the pairing culture and disk diffusion methods. Five isolated CFB showed $CaCO_3$ precipitation and antifungal activities against deteriorative fungal strains. Subsequently, five candidate bacteria were identified using 16S rDNA sequence analysis. Crack remediation, fungi growth inhibition, and water permeability reduction of antifungal CFB-treated cement surfaces were tested. All antifungal CFB showed crack remediation abilities, but only three strains (KNUC2100, 2103, and 2106) reduced the water permeability. Furthermore, these three strains showed fungi growth inhibition. This paper is the first application research of CFB that have antifungal activity, for an eco-friendly improvement of construction materials.

• KSBB Journal
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• v.32 no.3
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• pp.206-211
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• 2017

The physical method used to prevent a landslide has the risk of environmental pollution. Calcite forming bacteria (CFB) have been received increasing attention as a novel and environmental friendly strategy for the soil improvement. In this study, we selected 11 CFB strains with high calcite production. We also examined survivability and calcite productivity of the strains under various stress conditions to select strains with high resistance to various stresses. Two strains was selected by environment stress. Sphingobacterium sp. KJ-32 and Viridibacillus arenosi B-25 precipitate calcite more than other strains at pH 5 and $15^{\circ}C$ respectively. Bio-consolidated soil cakes were made using various calcium salts (calcium chloride, calcium acetate, calcium lactate, calcium gluconate) with mixed culture of 2 strains. Among them, the calcite made using calcium chloride was the largest. These observations demonstrate that this bio-consolidation technology has the potential for eco-friendly prevention of landslide and soil improvement.

• KSBB Journal
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• v.29 no.3
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• pp.188-192
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• 2014

Calcite forming bacteria (CFB) have been received increasing attention as a novel and environmental friendly strategy for the healing of concrete crack. Among the CFB, spore forming bacteria were proposed to overcome concrete condition (high pH, hydration heat, deicer). In this study, Lysinibaclillus sphaericus WJ-8 (WJ-8) isolated from concrete pavement was characterized. The WJ-8 was able to precipitate calcite at 10 mg/mL. When observed by scanning electron microscopy, WJ-8 showed spore formation and maximum spore yield was approximately 97.9%. Also response of spores against various environment stresses was examined. Approximately 83~97% of spores maintained their survivability at each three conditions ($60^{\circ}C$, 3 M NaCl and pH 12).

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.782-788
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• 2010

Microbiological calcium carbonate precipitation (MCP) has been investigated for its ability to improve the compressive strength of mortar. However, very few studies have been conducted on the use of calcite-forming bacteria (CFB) to improve compressive strength. In this study, we discovered new bacterial genera that are capable of improving the compressive strength of mortar. We isolated 4 CFB from 7 environmental concrete structures. Using sequence analysis of the 16S rRNA genes, the CFB could be partially identified as Sporosarcina soli KNUC401, Bacillus massiliensis KNUC402, Arthrobacter crystallopoietes KNUC403, and Lysinibacillus fusiformis KNUC404. Crystal aggregates were apparent in the bacterial colonies grown on an agar medium. Stereomicroscopy, scanning electron microscopy, and X-ray diffraction analyses illustrated both the crystal growth and the crystalline structure of the $CaCO_3$ crystals. We used the isolates to improve the compressive strength of cement-sand mortar cubes and found that KNUC403 offered the best improvement in compressive strength.

• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.1015-1020
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• 2012

Antifungal cement mortar or microbiological calcium carbonate precipitation on cement surface has been investigated as functional concrete research. However, these research concepts have never been fused with each other. In this study, we introduced the antifungal calcite-forming bacteria (CFB) Bacillus aryabhattai KNUC205, isolated from an urban tunnel (Daegu, South Korea). The major fungal deteriogens in urban tunnel, Cladosporium sphaerospermum KNUC253, was used as a sensitive fungal strain. B. aryabhattai KNUC205 showed $CaCO_3$ precipitation on B4 medium. Cracked cement mortar pastes were made and neutralized by modified methods. Subsequently, the mixture of B. aryabhattai KNUC205, conidiospore of C. sphaerospermum KNUC253, and B4 agar was applied to cement cracks and incubated at $18^{\circ}C$ for 16 days. B. aryabhattai KNUC205 showed fungal growth inhibition against C. sphaerospermum. Furthermore, B. aryabhattai KNUC205 showed crack remediation ability and water permeability reduction of cement mortar pastes. Taken together, these results suggest that the $CaCO_3$ precipitation and antifungal properties of B. aryabhattai KNUC205 could be used as an effective sealing or coating material that can also prevent deteriorative fungal growth. This study is the first application and evaluation research that incorporates calcite formation with antifungal capabilities of microorganisms for an environment-friendly and more effective protection of cement materials. In this research, the conception of microbial construction materials was expanded.

• Journal of Life Science
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• v.25 no.2
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• pp.237-242
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• 2015

The use of calcite-forming bacteria (CFB) in crack remediation and durability improvements in construction materials creates a permanent and environmentally-friendly material. Therefore, research into this type of application is stimulating interdisciplinary studies between microbiology and architectural engineering. However, the mechanisms giving rise to these materials are dependent on calcite precipitation by the metabolism of the CFB, which raises concerns about possible hazards to cement-based construction due to microbial metabolic acid production. The aim of this study was to determine target microorganisms that possibly can have bio-corrosive effects on cement mortar and to assess multi-functional CFBs for their safe application to cement structures. The chalky test was first used to evaluate the $CaCO_3$ solubilization feature of construction sites by fungi, yeast, bacterial strains. Not all bacterial strains are able to solubilize $CaCO_3$, but C. sphaerospermum KNUC253 or P. prolifica KNUC263 showed $CaCO_3$ solubilization activity. Therefore, these two strains were identified as target microorganisms that require control in cement structures. The registered patented strains Bacillus aryabhatti KNUC205, Arthrobacter nicotianae KNUC2100, B. thuringiensis KNUC2103 and Stenotrophomonas maltophilia KNUC2106, reported as multifunctional CFB (fungal growth inhibition, crack remediation, and water permeability reduction of cement surfaces) and isolated from Dokdo or construction site were unable to solubilize $CaCO_3$. Notably, B. aryabhatti KNUC205 and A. nicotianae KNUC2100 could not hydrolyze cellulose or protein, which can be the major constituent macromolecules of internal materials for buildings. These results show that several reported multi-functional CFB can be applied to cement structures or diverse building environments without corrosive or bio-deteriorative risks.