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Acid resistance is critical for the survival of bacteria in the dental caries oral micro-environment. However, there are few acid-resistant genes of microbiomes obtained through traditional molecular biology experimental techniques. This study aims to try macrogenomics technologies to efficiently identify acid-resistant genes in oral microbes of patients with dental caries. Total DNA was extracted from oral microbiota obtained from thirty dental caries patients and subjected to high-throughput sequencing. This data was used to build a metagenomic library, which was compared to the sequences of two S treptococcus mutant  known acid-resistant genes,  danK  and  uvrA , using a BLAST search. A total of 19 and 35 unknown gene sequences showed similarities with  S. mutans uvrA  and  dnaK  in the metagenomic library, respectively. Two unknown genes,  mo-dnaK  and  mo-uvrA , were selected for primer design and bioinformatic analysis based on their sequences. Bioinformatics analysis predicted them encoding of a human heat-shock protein (HSP) 70 and an ATP-dependent DNA repair enzyme, respectively, closely related with the acid resistance mechanism. After cloning, these genes were transferred into competent  Escherichia coli  for acid resistance experiments.  E. coli  transformed with both genes demonstrated acid resistance, while the survival rate of  E. coli  transformed with  mo-uvrA  was significantly higher in an acidic environment (pH = 3). Through this experiment we found that identify unknown acid-resistant genes in oral microbes of patients with caries by establishing a metagenomic library is very efficient. Our results provide an insight into the mechanisms and pathogenesis of dental caries for their treatment without affecting oral probiotics.   Supplementary Information  The online version contains supplementary material available at 10.1186/s13568-021-01199-4.

Identification of unknown acid-resistant genes of oral microbiotas in patients with dental caries using metagenomics analysis