Thirty bacterial isolates were tested on three different media for Poly-ß-hydroxybutyrate (PHB) production. The best bacterial isolates for producing PHB were screened and identified based on molecular biology; then, using three different alternative carbon sources (dried whey, sugar beet molasses and date molasses), physical properties were evaluated by Infrared (IR) spectrometry and Gas chromatography-mass spectrometry (GC-MS/MS) analysis. Our results showed that the best isolates identified based on molecular biology were Bacillus paramycoides MCCC 1A04098, Azotobacter salinestris NBRC 102611 and Brevundimonas naejangsanensis BIO-TAS2-2. The addition of sugar beet molasses to the medium of A. salinestris increased the cell dry weight (CDW), PHB concentration, PHB% and conversion coefficient (4.97 g/L, 1.56 g/L, 31.38% and 23.92%, respectively). The correlation coefficient values between PHB g/L and CDW g/L varied between very strong and moderate positive correlation. IR of the produced PHB from B. paramycoides and A. salinestris showed similar bands which confirmed the presence of PHB; however, B. naejangsanensis showed weak bands, indicating lower PHB concentration. The chemical composition obtained showed that the GC-MS of the PHB extracted represents 2, 4-ditert-butylphenol for B. paramycoides and isopropyl ester of 2-butenoic acid for both of A. salinestris and Brevundimonas naejangsanensis. Therefore, PHB produced by microorganisms can be considered a biodegradable polyester, and represents a promising technique for the development of eco-friendly and fully biodegradable plastics.
Keywords: GC-MS/MS spectrometry; IR spectrometry; PHB; agro-industrial waste.