Functional analysis of a putative holin-like peptide-coding gene in the genome of Bacillus licheniformis AnBa9.
BhlA, a putative holin-like protein of Bacillus licheniformis AnBa9 expressed in Escherichia coli BL21(DE3) showed antibacterial activity against several gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and Micrococcus luteus. Deletion analysis of bhlA suggests that a hydrophobic transmembrane domain, BhlATM is essential for antibacterial activity. Though the minimum inhibitory concentration (MIC) of BhlA was sevenfold lower than BhlATM, transmembrane domain deleted construct (BhlATM) had no antibacterial activity. The expression of BhlA in E. coli was found to be toxic to cells. Therefore, the bhlA was cloned in yeast surface display vector pYD1 and expressed in Saccharomyces cerevisiae. The surface displayed yeast showed inhibition of several gram-positive bacteria. This recombinant yeast expressing BhlA may be used as biodrug for efficient control of multiple drug-resistant bacterial infections.
Anthony
Thangamani
T
Department of Genetics, School of Biological Sciences, Centre for Excellence in Genomic Sciences, Madurai Kamaraj University, Madurai, India.
Chellappa
Gunasekaran Stalin
GS
Rajesh
Thangamani
T
Gunasekaran
Paramasamy
P
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000145:classification / Q000378:metabolism* / Q000494:pharmacology; D001407:Bacillus / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D019149:Bioreactors; D002463:Cell Membrane Permeability / Q000187:drug effects; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D006094:Gram-Positive Bacteria / Q000187:drug effects; D008565:Membrane Proteins / Q000378:metabolism; D016106:Methicillin Resistance / Q000187:drug effects; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010455:Peptides / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013204:Staphylococcus Phages / Q000378:metabolism; D001618:beta-Lactamases / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D008565:Membrane Proteins; D010455:Peptides; D001618:beta-Lactamases