Metagenomic and mineralogical insights into biomineralization of shell lime putty prepared with fermented Terminalia chebula fruit extract for built heritage applications

The study investigates an ancient Indian fermentation technique using 10% w/v Terminalia chebula fruit extract (K10%) and its interaction with shell lime. Fermentation ages of 1, 4, 7, and 14 days of K10% were identified as key stages based on carbon dioxide evolution rates, with peaks at 4 and 7 days (8826.67 ppm/hour and 6923.32 ppm/hour, respectively). These stages were further investigated via 16S rRNA (V3–V4 region) metagenomic sequencing, identifying dominant microbial taxa including Acinetobacter guillouiae , Bacillus safensis , Enterobacter cowanii , Lactobacillus helveticus, Lactobacillus casei , and Sphingobacterium multivorum . These strains facilitated biomineralization through carbonic anhydrase activity, supported by exopolysaccharide production and hydrolytic enzymes, in putty variants prepared with a K10%-to-binder mass ratio of 0.95. Gas chromatography–mass spectrometry profiling of the putty extracts revealed a progression of unsaturated fatty acids and an increase in ester content, particularly in the variant prepared with 4-day aged K10%. XRD analysis at 90 days confirmed the highest levels of calcite (64.4% and 60.17%) and tobermorite (5.27% and 5.14%) in the lime putty variants prepared with 4- and 7-day aged K10%. These variants also confirmed elevated carbonation levels (53.36% and 51.95%), as determined by thermogravimetric analysis. Mineralogical developments contributed to enhanced carbonation, matrix densification, and reduced water permeability, corroborated by superior mechanical and other durability parameters compared to the reference putty. Overall, the findings demonstrate that K10% extract significantly enhances the functional and durability characteristics of lime binders, offering a bio-based, sustainable approach suitable for internal and external applications in heritage restoration and eco-friendly construction. • Use of fermented T.chebula fruit in lime binders is a sustainable technique of lime binder preparation in Indian heritage. • K10% w/v fermented extract at different ages showed erratic CO 2 evolution rates, affecting the mineralogical and carbonation characteristics of lime putty. • V3 and V4 Metagenomic analysis confirmed the enzymatic route of biomineralization activity in the lime putty supported by GC-MS, XRD, FT-IR, and TGA analysis.