Functional cloning and predictive structural modeling of a novel esterase from Bacillus subtilis strain, RRL 1789.
We have recently reported the purification and characterization of a novel esterase from the Bacillus subtilis strain. In the present study we report the genomic DNA cloning and predictive structural modeling of this novel esterase. Tributyrin- and Rhodamine B-based functional screen of a Bacillus subtilis genomic library led to the identification of a potential lipolytic gene. DNA sequence analysis of the cloned gene showed that it encodes a protein of 489 amino acid residues. Sequence homology search and multiple sequence alignment showed that the protein was highly homologous to known esterases. Secondary structure-driven multiple sequence alignment with the homologous esterase of known three-dimensional structures was performed and a 3D structure model of this enzyme was constructed. Based on the topological organization of the secondary structures, this protein belongs to the alpha/beta hydrolase superfamily. Moreover, the presence of serine in the context of amino acid sequence G/A-X-S-X-G (with X an arbitrary amino acid residue) in the protein indicates that it belong to the class of serine hydrolases of this superfamily.
Peerzada
Kaiser
K
Department of Biotechnology, University of Kashmir, Hazratbal, Srinagar, Kashmir, India.
Ul Hussain
Mahboob
M
Jan
Nishawar
N
Verma
Vijeshwar
V
Qazi
Ghulam N
GN
Andrabi
Khurshid I
KI
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D004950:Esterases / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D008958:Models, Molecular; D008969:Molecular Sequence Data; D012329:RNA, Bacterial / Q000032:analysis; D016415:Sequence Alignment
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D004950:Esterases