Calcium carbonate precipitation induced by a bacterium strain isolated from an oligotrophic cave in Central China

Abstract

A heterotrophic Bacillus sp. strain (5C-1) was isolated from Heshang cave, an oligotrophic karst cave in the middle reaches of Yangtze River, and identified by BIOLOG and 16S rDNA sequencing. Bacterially induced formation of calcium carbonate by 5C-1 was investigated in several comparative experimental sets with or without the cell and extracellular enzymes. The temporal variations of both the amount of the precipitates and the pH values of the solution were measured by a spectrophotometer and a pH meter, respectively. The morphological characteristics of the calcium carbonate precipitates were observed with environmental scanning electronic microscopy (ESEM). The growth of 5C-1 was found to greatly promote the pH value of the liquid medium in the first 2 days, which favors the formation of calcium carbonate. No precipitates were formed with the pH value lower than 8.6, though the pH value was demonstrated to be not the only factor controlling the formation of the calcium carbonate. The accumulation of extracellular polysaccharide substance was observed to favor the precipitate formation. Only when both factors reached a threshold did the precipitates form with the addition of CaCl₂. Cells and extracellular enzymes were not the factors that limit the precipitate formation in our microbial systems. The precipitates of a variety of morphological features including dumb bells, peanuts, irregular and spherical and rhombic forms were mainly observed in our microbial systems but not in the chemical control system. Interestingly, imprints of bacterial cells and spores were observed to be present on the surface of the precipitates of a peanut or a dumb bell form, probably indicative of the microbial escaping mechanism during the mineralization of calcium carbonate.

Keywords

Bacillus / oligotrophic karst cave / calcium carbonate / bacterially induced mineral formation