The viability of spores is the key factor for microbial induced calcium carbonate precipitation.

While previous studies mainly focused on the total number of spores as an index to predict the calcium precipitation activity (CPA) of bacterial strains, the effect of viability of spores on microbial-induced calcium precipitation (MICP) has remained highly ignored. Therefore, for the first time, we have attempted to optimize the sporulation process in terms of viable spore production and, most importantly, aimed to build a correlation between viable spores and CPA. The results have shown that for the sporulation of Bacillus sp. H4, starch and peptone are the optimal carbon and nitrogen sources, respectively. One gram per liter of sodium chloride promotes CPA and production of viable spores, whereas an increase of sodium chloride concentration beyond 8 g L-1 significantly reduces CPA without reducing the quantity of viable spores. Exogenous conditions such as seed age, inoculation quantity, and liquid volume only pose slight influence on the sporulation and CPA. Conclusively, the spores produced under optimized conditions are more morphologically uniform and display a 20% increase in CPA compared to pre-optimized spores. Furthermore, by combining the results of heatmap analysis, it can be concluded that not only the quantity, but also the quality of viable spores is important for bacterial strain to develop high CPA and effective MICP process. This study sheds light on the breadth of biomineralization activity based on viable spores and is an imperative step toward the intelligible design of MICP-based engineering solutions. KEY POINTS: • Viability of spores is a key controlling factor in calcium precipitation activity (CPA). • Spores produced under optimized conditions display a 20% increase in CPA. • Quality of viable spores is imperative for bacterial strains to develop high CPA.