• P.NARMATHA DEVI
• A.MAHALAKSHMI
• K.SUJATHA

Polyhydroxyalkanoates (PHAs) are considered as very good alternative to synthetic plastics not only due to their similar physico-chemical properties but it can be degraded into CO2 and H2O in the environment by either intra or extracellular enzymatic (PHA depolymerase -phaZ) activity of PHA degrading bacteria. In intracellular degradation, accumulated PHA is mobilized by the bacteria in stressed environmental conditions for survival, whereas in extracellular degradation, enzyme will act on PHA released into the environment after cell lysis. The existing technique used to screen PHA degrading bacteria is observation of clear zone formation, as this method is influenced by polymer preparation, microbial culture and long incubation time (3-5days), there is a need to design an effective simple screening technique to select positive isolates from the environmental sample. Hence, in this study, specific primer pair was designed to amplify ~800 bp conserved region of mcl phaZ harbouring microbial isolates from environmental sample. The ideal colony/lysate PCR parameter was ascertained after repeated trials with reference strain Pseudomonas putida (MTCC-9782). Strains were isolated from environmental sample, the primer specificity was evaluated by sequencing randomly selected PCR positive phaZ strain (labelled as ND1). Similarity analysis of the phaZ and 16S rDNA identified the strain as Pseudomonas sp. PHA accumulation and degradation ability of the strain was confirmed further using FT-IR and esterase activity respectively. Therefore, in future, this phaZ specific primer provides a quick, powerful and reliable alternative to conventional method in the detection and identification of PHA degrading bacteria.

Polyhydroxyalkanoates, PHA depolymerase, Colony PCR, FT-IR, PHA synthase

"PCR VALIDATION OF GENE SPECIFIC PRIMER TO DETECT INTRACELLULAR MCL POLYHYDOXYALKANOATE DEGRADING BACTERIA FROM ENVIONMENTAL SAMPLE", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.6, Issue 3, page no.483-490, March-2019, Available :http://www.jetir.org/papers/JETIR1903973.pdf

"PCR VALIDATION OF GENE SPECIFIC PRIMER TO DETECT INTRACELLULAR MCL POLYHYDOXYALKANOATE DEGRADING BACTERIA FROM ENVIONMENTAL SAMPLE", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.6, Issue 3, page no. pp483-490, March-2019, Available at : http://www.jetir.org/papers/JETIR1903973.pdf