Bioinformatics analysis of Bacillus subtilis isoprene enzyme ComQ and its effect on biofilm morphology

Abstract:

[Objective] Bacillus subtilis ComQ is a kind of isoprene biosynthesis enzyme. We used bioinformatics to predict and analyze the biological characteristics of ComQ. In addition, we also constructed comQ overexpression and knockout mutants to verify their effect on biofilm morphology. [Methods] Firstly, we obtained the amino acid sequence of ComQ protein from the Protein database in the NCBI website. Secondly, we predicted and analyzed the biological characteristics including physical and chemical properties, signal peptide, domain and spatial structure of ComQ by online bioinformatics software. Thirdly, we constructed comQ overexpression and knockout strain of Bacillus subtilis, and observe their growth characteristics of biofilm by plate fermentation. [Results] ComQ protein was composed of 299 amino acids with a theoretical molecular weight of about 34 204.08 Da. It was a stable intracellular protein without signal peptide and transmembrane region. Colony PCR verified that Bacillus subtilis BS168 mutants were constructed successfully. The biofilm morphology of the two mutants was different from that of the original strain BS168. [Conclusion] Through the prediction and analysis of the basic properties, key amino acid sites and protein structure of ComQ protein, the mutant strain was constructed to verify the morphological changes of biofilm, which laid a foundation for the subsequent study of ComQ on the growth and metabolism of Bacillus subtilis.