Screening and Production of Biodegradable Polyester Poly(3-hydroxybutyrate) by Bacteria Endophytic to Brassica nigra L.

Abstract

Aims: Poly(3-hydroxybutyrate) [P(3HB)], the microbially produced biodegradable thermoplastics has find wide range of applications in recent years. Development of low cost production strategies utilizing novel organisms is a crucial challenge. Present study is aimed to isolate and screen bacterial endophytes of Brassica nigra L. for the production of P(3HB). Place and Duration of Study: The experiments were performed in the Microbiology Laboratory, Department of Botany, University of Calcutta, Kolkata during 2012-2014. Methodology: Culturable bacterial endophytes were isolated from surface sterilized healthy tissues of B. nigra L. and screened for P(3HB) production in mineral salts medium. The chloroform extracted dried polymer was treated with H2SO4 and quantified spectrophotometrically at 235 nm. Results: About 78% of the bacterial endophytes recovered from surface sterilized B. nigra L. tissues showed different degrees of P(3HB) accumulation. Isolates (9) showing P(3HB) accumulation exceeding 10% of the cell dry weight (CDW) were characterized and tentatively identified as members of Bacillus, Pseudomonas, Xanthomonas, Alcaligenes and Acetobacter. The most potent isolate, BNL 06 identified as Bacillus pumilus BNL 06 (GenBank Accession No. KP202723), accumulated P(3HB) accounting 18% of CDW with an yield of 0.55 g/l. Finally the nature of the polymer was further confirmed by FTIR analysis. Conclusion: Exploration of the endophytic bacterial diversity of B. nigra L. have clearly revealed the potential of Bacillus pumilus BNL 06 for P(3HB) production as an alternative source of thermoplastics.