[국내논문]Calcium Carbonate Precipitation by Bacillus and Sporosarcina Strains Isolated from Concrete and Analysis of the Bacterial Community of Concrete원문보기
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 (...
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.
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.
Nonetheless, the reasons for mineralization by CCP-capable bacteria, regulation of the possible CCP-specific genetic system, and the role or effects of CCP-capable bacteria in ecosystems are still poorly understood. The purpose of this study was to isolate and characterize novel CCP-capable bacteria from concrete. We sampled concrete from two locations (Jingyo and Seoul, South Korea) and characterized them by field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectrometry (EDS) mapping, urease activity assays, and EPS quantification.
EPS quantification was conducted according to previous studies, with slight modifications [25]. One milliliter of cell culture was mixed with Congo red at the final concentration of 3.
대상 데이터
Taxonomic affiliation was determined by sequencing the 16S rRNA and by construction of phylogenetic trees via sequence similarity by means of EzTaxon [6]. The 16S rRNA sequences of the three isolates have been deposited in National Center for Biotechnology Information (NCBI) GenBank under the accession numbers KU168425, KU168426, KU168427 for strains JH3, JH7, and HYO08, respectively. The optimal liquid medium for strains JH3 and JH7 was Luria-Bertani broth, and for strain HYO08 was tryptic soy broth with 2% urea (w/v).
이론/모형
Urease activity was measured by a colorimetric assay [30]. Briefly, the strains JH3, JH7, and HYO08 and Escherichia coli MG1655 (negative control) were grown to the stationary phase.
A biofilm formation assay was conducted by the crystal violet staining method [16]. Briefly, the four strains, including Escherichia coli MG1655 as a control, were incubated overnight with aeration and washed twice with PBS.
성능/효과
Recent studies suggest that urea is abundant in CCP environments because of input from various sources: mammal urine, water infiltration from the surface, human activities, and agricultural waste [19,31]. Therefore, these findings suggest that the concrete samples from which we isolated the CCP-capable bacteria might have contained urea that the isolates could use for metabolism and formation of minerals.
The results of community analysis also showed that the relative abundance of Bacillus and Sporosarcina was low, even though we successfully isolated those strains. This discrepancy could be due to a culturing bias that possibly provided favorable conditions for Bacillus and Sporosarcina, or due to our focused screening strategy targeting alkali-tolerant strains.
후속연구
S5). Although the nucleation effect of CCP-capable bacteria is still a hypothesis [3,13], the exact location triggering CCP in our three strains needs further research.
The presence of Cyanobacteria and free-living nitrogen-fixing Actinobacteria such as Arthrobacter and Propionibacterium may play an important role by supplying nutrients to a concrete-dwelling community because an endolithic lifestyle involves extreme starvation [15,38]. For practical applications of CCP-capable bacteria, the relation between the capacity for CCP and other physiological features of CCP-capable bacteria as well as the interaction of CCP-capable bacteria with other species should be studied in more detail.
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