Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 9 (1), 49

Enhanced Calcium Carbonate-Biofilm Complex Formation by Alkali-Generating Lysinibacillus Boronitolerans YS11 and Alkaliphilic Bacillus Sp. AK13

Affiliations

Enhanced Calcium Carbonate-Biofilm Complex Formation by Alkali-Generating Lysinibacillus Boronitolerans YS11 and Alkaliphilic Bacillus Sp. AK13

Yun Suk Lee et al. AMB Express.

Abstract

Microbially induced calcium carbonate (CaCO3) precipitation (MICP) is a process where microbes induce condition favorable for CaCO3 formation through metabolic activities by increasing the pH or carbonate ions when calcium is near. The molecular and ecological basis of CaCO3 precipitating (CCP) bacteria has been poorly illuminated. Here, we showed that increased pH levels by deamination of amino acids is a driving force toward MICP using alkalitolerant Lysinibacillus boronitolerans YS11 as a model species of non-ureolytic CCP bacteria. This alkaline generation also facilitates the growth of neighboring alkaliphilic Bacillus sp. AK13, which could alter characteristics of MICP by changing the size and shape of CaCO3 minerals. Furthermore, we showed CaCO3 that precipitates earlier in an experiment modifies membrane rigidity of YS11 strain via upregulation of branched chain fatty acid synthesis. This work closely examines MICP conditions by deamination and the effect of MICP on cell membrane rigidity and crystal formation for the first time.

Keywords: Alkaline generation; Bacteria-CaCO3 interaction; Branched chain fatty acid synthesis; Dual species CaCO3 precipitation; Membrane rigidity.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
a Circular genomic map of Lysinibacillus boronitolerans YS11. From the outside to the center are RNA genes, genes of the reverse strand, genes on the forward strand, GC ratio, and GC skew. Genes are colored according to their COG category. b Number of CDSs belonging to amino acid metabolism and transport group from YS11 with other Lysinibacillus species including L. boronitolerans NBRC 103108T, L. macroides DSM 54 T, L. xylanilyticus DSM 23493 T, and L. pakistanensis JCM 18776 T. c COG category of Lysinibacillus species. d Quantification of CDSs involved in deamination of amino acids among amino acid metabolism and transport groups from Lysinibacillus species
Fig. 2
Fig. 2
a Quantification of ammonia production from supernatant of YS11 cultured in YL medium. b pH increases by YS11 in amino acid rich media (.8% yeast extract or .8% nutrient broth). c Expression of ammonia releasing genes from YS11 that exhibited RPKM value higher than 100
Fig. 3
Fig. 3
Growth, pH changes, and calcium utilization in MICP-inducing conditions with YS11, AK13 alone, and YS11 + AK13 coculture. a Based on CFU growth of YS11, AK13, and YS11 + AK13, the growth of AK13 is facilitated by coculture with YS11. b pH changes in YS11, AK13 single culture, and YS11 + AK13 coculture are shown. c Unbound Ca2+ concentrations in supernatants of YS11, AK13 single culture, and YS11 + AK13 coculture are measured. d Induced growth of AK13 from coculture with YS11 in agar plate cultures
Fig. 4
Fig. 4
a Field emission scanning electron microscopy (FE-SEM) of calcium carbonate precipitated by YS11 single culture and YS11 + AK13 coculture (×30,000 and ×5000 magnification). b FTIR analysis of precipitated calcium carbonate. c CLSM image of biofilm and calcium carbonate produced by YS11 single culture. d Crystal violet quantification of biofilm formation in single culture or YS11 + AK13 coculture. e Killing curve assay of YS11 alone, AK13 alone, and YS11 plus AK13 under pH 12
Fig. 5
Fig. 5
a Growth curve of YS11 in calcium-rich condition (CaAc) and calcium-poor condition (NaAc). b Schematic view of upregulated branched chain amino acid (BCAA) and branched chain fatty acid (BCFA) synthesis in CaAc. c Modified BCFA ratio in CaAc compared to that in NaAc
Fig. 6
Fig. 6
a Enhanced spore formation of YS11 in calcium-rich condition. b Increased biofilm formation in calcium-rich condition. c CSLM image of increased biofilm (red) formation in calcium-rich condition (blue color indicates calcium carbonate mineral). d Increase in swimming motility in calcium-rich condition compared to that in calcium-poor condition

Similar articles

See all similar articles

References

    1. Andrews SC, Robinson AK, Rodriguez-Quinones F. Bacterial iron homeostasis. FEMS Microbiol Rev. 2003;27:215–237. doi: 10.1016/S0168-6445(03)00055-X. - DOI - PubMed
    1. Arp G, Reimer A, Reltner J. Photosynthesis-induced biofilm calcification and calcium concentrations in Phanerozoic oceans. Science. 2001;292:1701–1704. doi: 10.1126/science.1057204. - DOI - PubMed
    1. Bai Y, Guo XJ, Li YZ, Huang T. Experimental and visual research on the microbial induced carbonate precipitation by Pseudomonas aeruginosa. AMB Express. 2017;7:57. doi: 10.1186/s13568-017-0358-5. - DOI - PMC - PubMed
    1. Bertrand JJ, West JT, Engel JN. Genetic analysis of the regulation of type IV pilus function by the Chp chemosensory system of Pseudomonas aeruginosa. J Bacteriol. 2010;192:994–1010. doi: 10.1128/JB.01390-09. - DOI - PMC - PubMed
    1. Bhaskar PV, Bhosle NB. Bacterial extracellular polymeric substance (EPS): a carrier of heavy metals in the marine food-chain. Environ Int. 2006;32:191–198. doi: 10.1016/j.envint.2005.08.010. - DOI - PubMed

LinkOut - more resources

Feedback