Alkaline-Tolerant Bacillus cereus 12GS: A Promising Polyhydroxybutyrate (PHB) Producer Isolated from the North of Mexico.

Environmental pollution caused by petroleum-derived plastics continues to increase annually. Consequently, current research is interested in the search for eco-friendly bacterial polymers. The importance of Bacillus bacteria as producers of polyhydroxyalkanoates (PHAs) has been recognized because of their physiological and genetic qualities. In this study, twenty strains of Bacillus genus PHA producers were isolated. Production was initially evaluated qualitatively to screen the strains, and subsequently, the strain B12 or Bacillus sp. 12GS, with the highest production, was selected through liquid fermentation. Biochemical and molecular identification revealed it as a novel isolate of Bacillus cereus. Production optimization was carried out using the Taguchi methodology, determining the optimal parameters as 30 °C, pH 8, 150 rpm, and 4% inoculum, resulting in 87% and 1.91 g/L of polyhydroxybutyrate (PHB). Kinetic studies demonstrated a higher production within 48 h. The produced biopolymer was analyzed using Fourier-transform infrared spectroscopy (FTIR), confirming the production of short-chain-length (scl) polyhydroxyalkanoate, named PHB, and differential scanning calorimetry (DSC) analysis revealed thermal properties, making it a promising material for various applications. The novel B. cereus isolate exhibited a high %PHB, emphasizing the importance of bioprospecting, study, and characterization for strains with biotechnological potential.

Utilization of Agave durangensis leaves by Bacillus cereus 4N for polyhydroxybutyrate (PHB) biosynthesis.

Lignocellulosic wastes may provide a means to economize polyhydroxybutyrate (PHB) production. This study has proposed the use of Agave durangensis leaves obtained from the artisanal mezcal industry as a novel substrate for this aim. Results revealed an increase in PHB biosynthesis (0.32 g/L) and improvement in %PHB (16.79-19.51%) by Bacillus cereus 4N when A. durangensis leaves used as carbon source were physically pre-treated by ultrasound for 30 min (ADL + US30') and thermally pre-treated (ADL + Q). Chemical analyses and SEM studies revealed compositional and morphological changes when A. durangensis leaves were physically pre-treated. Also, elemental analysis of growth media showed that carbon/nitrogen ratios of 14-21, and low nitrogen, hydrogen, and protein content were well-suited for PHB biosynthesis. Confocal microscopy revealed morphological changes in the bacterial cell and carbonosome structure under the influence of different substrates. Finally, Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) analyses showed that homopolymeric PHB with a high thermal-resistance (271.94-272.89 °C) was produced. Therefore, the present study demonstrates the potential use of physically pre-treated A. durangensis leaves to produce PHB. These results promote the development of a circular economy in Mexico, where lignocellulosic wastes can be employed to produce value-added biotechnological products.

Efficient recovery of thermostable polyhydroxybutyrate (PHB) by a rapid and solvent-free extraction protocol assisted by ultrasound.

Nowadays, there are great research interest in polyhydroxybutyrate (PHB) recovery protocols that reduce the use of organic solvents and efficiently recover this bacterial biopolymer. The present study reports an extraction protocol assisted by ultrasound, which is a rapid protocol that increases the amount of polymeric matter extracted, reduces the cellular digestion step with sodium hypochlorite and eliminates the use of organic solvents. Likewise, characterization studies by Fourier transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance spectroscopy (1H NMR), differential scanning calorimetry (DSC), and X-ray diffraction (XRD) demonstrated that the biopolymer recovered by this protocol is PHB homopolymer with a major thermal-resistance and crystalline properties. Besides, in this study are reported the thermic and crystalline differences between the PHB obtained from the fermentation of complex carbohydrates (agavins) and simple sugars. The biopolymer obtained by this rapid extraction protocol would be suitable for ecological and biomedical applications, due to the low melting temperature, less than 50% crystallinity, and the lack of lipopolysaccharides. Therefore, this extraction protocol might represent an alternative to the traditional protocol based on NaOCl-chloroform and is part of the green trend to improve the PHB production.