Production and Characterization of Poly(3-hydroxybutyrate) by Thermophilic Geobacillus sp. Strain SL28 Isolated from a Natural Hot Spring

Abstract

Polyhydroxyalkanoates (PHAs) are biodegradable microbial polyesters that may reduce dependence on petroleum-derived plastics, but broader use remains constrained by production costs and tightly controlled cultivation. Thermophilic microorganisms have attracted increasing attention as promising PHA producers because they can be cultivated at elevated temperatures, reducing contamination risks and operational costs. In this study, a thermophilic Geobacillus sp. strain SL28 isolated from a Vietnamese hot spring was evaluated for its PHA-producing capability. Temperature, initial pH, carbon and nitrogen sources, C/N ratio, and cultivation time were examined. Intracellular inclusions were assessed by Sudan Black B and Nile Blue A staining and transmission electron microscopy, whereas the recovered polymer was characterized by FE-SEM, Fourier-transform infrared spectroscopy (FTIR), gas chromatography–mass spectrometry (GC–MS), 1H- and 13C-nuclear magnetic resonance (NMR) spectroscopy, capillary viscometry, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and tensile testing. Under the optimized cultivation conditions identified by one-factor experimentation (50 °C, pH 7.0, sucrose, peptone, C/N 25:1, and 72 h), SL28 reached a dry cell weight of 2.215 ± 0.16 g/L and a PHA concentration of 1.383 ± 0.04 g/L, equivalent to 62.57 ± 2.82% of DCW. Structural characterization confirmed that the recovered polymer was poly(3-hydroxybutyrate) (PHB). The purified PHB also exhibited favorable thermal stability, a relatively high molecular weight, and satisfactory mechanical properties. These results demonstrate the potential of thermophilic Geobacillus sp. SL28 as a promising candidate for PHB production.