This review summarizes the molecular mechanisms of biofilm formation and disassembly in *Bacillus subtilis*, a non-pathogenic Gram-positive bacterium. Biofilms are communities of bacteria enclosed in an extracellular matrix, and *B. subtilis* has become an important model organism for studying these processes. The review highlights the complex regulatory pathways that control biofilm assembly, including the Spo0A pathway, which is central to matrix gene expression and sporulation. The matrix is primarily composed of exopolysaccharides (EPS) and proteins, and its structure and function are crucial for biofilm integrity. The review also discusses the role of various proteins, such as TasA and BslA, in biofilm formation and the regulatory mechanisms that control their expression. The review outlines the regulatory pathways that govern biofilm formation, including the SlrR-SinR epigenetic switch, which controls the expression of matrix genes. The Spo0A pathway is regulated by phosphorylation and interacts with other regulatory proteins to control gene expression. The review also describes the role of kinases such as KinC and KinD in triggering matrix gene expression, as well as the role of cannibalism in increasing the population of matrix-producing cells. The review discusses the mechanisms by which *B. subtilis* disassembles biofilms, including the release of D-amino acids and norspermidine, which disrupt the extracellular matrix and lead to biofilm dispersal. The review also explores the potential of small molecules, such as D-amino acids and polyamines, as biofilm inhibitors that could be used to control biofilm formation in other bacterial species. These findings highlight the importance of understanding the molecular mechanisms of biofilm formation and disassembly for developing strategies to control biofilms in both medical and industrial settings.This review summarizes the molecular mechanisms of biofilm formation and disassembly in *Bacillus subtilis*, a non-pathogenic Gram-positive bacterium. Biofilms are communities of bacteria enclosed in an extracellular matrix, and *B. subtilis* has become an important model organism for studying these processes. The review highlights the complex regulatory pathways that control biofilm assembly, including the Spo0A pathway, which is central to matrix gene expression and sporulation. The matrix is primarily composed of exopolysaccharides (EPS) and proteins, and its structure and function are crucial for biofilm integrity. The review also discusses the role of various proteins, such as TasA and BslA, in biofilm formation and the regulatory mechanisms that control their expression. The review outlines the regulatory pathways that govern biofilm formation, including the SlrR-SinR epigenetic switch, which controls the expression of matrix genes. The Spo0A pathway is regulated by phosphorylation and interacts with other regulatory proteins to control gene expression. The review also describes the role of kinases such as KinC and KinD in triggering matrix gene expression, as well as the role of cannibalism in increasing the population of matrix-producing cells. The review discusses the mechanisms by which *B. subtilis* disassembles biofilms, including the release of D-amino acids and norspermidine, which disrupt the extracellular matrix and lead to biofilm dispersal. The review also explores the potential of small molecules, such as D-amino acids and polyamines, as biofilm inhibitors that could be used to control biofilm formation in other bacterial species. These findings highlight the importance of understanding the molecular mechanisms of biofilm formation and disassembly for developing strategies to control biofilms in both medical and industrial settings.