Abstract

Background: Collagenase is a type of proteolytic enzyme that specifically hydrolyzes native collagen with a three-dimensional helical structure. The structure and properties of collagenase vary with different sources and types. In addition to the well-characterized Clostridium collagenase, other collagenases are largely unknown. Various gene and protein databases have been widely used to mine novel functional genes in the genome. Gene mining and sequence analysis are effective methods for studying these enzymes. Objective: The present study aimed to understand the molecular, structural, and functional characteristics of collagenase from Bacillus cereus MH19 using a bioinformatics approach. Method: Based on the three-generation sequencing PacBio technique, Sequencing the Bacillus cereus MH19 genome. Function annotation is completed by blasting genes with different databases. Collagenases were investigated based on the physiochemical properties, phylogenetic relation, and domain architecture. The 3D structure model of the selected collagenase has been constructed and verified by SAVES. Results: There were 5 collagenases in Bacillus cereus MH19 with a molecular weight distribution ranging from 36-110 kDa. The analysis of evolutionary relationship between different collagenases indicating that the BCC000504 and BCC003388 collagenase gene sequences are closer to the typical collagenase genes ColG, ColA, and ColH, followed by BCC003615. The domain and function analysis showed that the collagenases BCC000504 and BCC003388 were similar to the collagenases ColG, ColA and ColH. BCC004271 was similar to BCC004272, and BCC003615 might be quite different from other collagenases. The secondary structure of collagenase was analyzed. The SAVES evaluation indicates that 3D structural modeling of the selected collagenase is acceptable. Conclusion: This study provides an overview of the molecular, functional, and structural characteristics of collagenase from Bacillus cereus MH19, which helps to understand the bacterial collagenase. The characterization of the collagenase will certainly expand the application range of collagenase.

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