Phylogenetic Analysis and Investigation of the Consensus Sequence of Alpha- Amylase-Producing Bacillus Species for Potential Amylase with Enhanced Activity

Alpha-amylase enzymes from Bacillus species are integral to various industrial processes due to their efficient starch hydrolysis capabilities. This study aimed to conduct a phylogenetic analysis to understand the evolutionary relationships between alpha-amylase-producing Bacillus species and investigate the consensus sequence derived from the multiple sequence alignment for its potential for alpha-amylase with enhanced activity. Sixteen (16) alpha-amylase protein sequences from Bacillus species were retrieved from the NCBI database and imported to Expasy server to calculate the physicochemical features. The Geneious Prime software was employed to perform multiple sequence alignment and construct a phylogenetic tree. The consensus sequence obtained from the MSA was analysed by determining its physicochemical features, predicting the functional domains, and predicting the 3D secondary structure. The physicochemical features of the sixteen sequences revealed a notable range of features, and the multiple sequence alignment revealed several conserved regions with consecutive identical residues, including highly conserved region sequence such as NGTLMQYFEW, conserved region sequences such as ENG, NHD, and less conserved region sequences such as DG, PL, LA, and YG. The consensus sequence analysis revealed physicochemical features, including 534 amino acids, molecular weight of 60,553.30 Da, theoretical pI of 5.47, negative residues (64), positive residues (49), instability index (24.92), aliphatic index (70.30), GRAVY (-0.537), and extinction coefficients (156650). The analysis also revealed functional domains like alpha-amylase and glycosyl hydrolase, and a 3D secondary structure prediction showed a stable conformation suitable for industrial applications. This study enhanced our understanding of the evolutionary and functional relationships among alpha-amylase- producing Bacillus species and the potential of utilising consensus sequences in enzyme engineering.