Phloroglucinol (PHL) or 2,4-diacetylphloroglucinol (DAPG) is a polyketide compound that exhibits broad spectrum anti-bacterial and antifungal properties against soil-borne plant pathogens. It is produced by gram negative bacterium Pseudomonas (fluorescent Pseudomonad) which renders them a potential biological control agent. The process of PHL production is accomplished by a well coordinated activity of four (phlABCD) genes arranged in an operon. All four genes are essential and indispensable for biosynthesis of DAPG from precursor malonyl CoA. Here, we report cloning and analysis of phlB gene from an Indian strain of Pseudomonas fluorescens. Further, we predicted the structure of PHLB protein and analyzed its intrinsic details. Stereo-chemical properties were analyzed by Ramachandran plot and secondary structure was retrieved by PDB sum. Our study provides insight into the structure of PHLB protein and revealed that though Pseudomonas PHLB lacks conserved motifs and domains it is indispensable for monoacetyl phloroglucinol (MAPG) synthesis.