Biosynthesis and biocompatibility evaluation of zinc oxide nanoparticles prepared using Priestia megaterium bacteria.

The current study aimed to find an effective, simple, ecological, and nontoxic method for bacterial green synthesis of zinc oxide nanoparticles (ZnONPs) using the bacterial strain Priestia megaterium BASMA 2022 (OP572246). The biosynthesis was confirmed by the change in color of the cell-free supernatant added to the zinc nitrate from yellow to pale brown. The Priestia megaterium zinc oxide nanoparticles (Pm/ZnONPs) were characterized using UV-Vis spectroscopy, high-resolution transmission electron microscopy (HR-TEM), energy-dispersive X-ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), and zeta potential. The Pm/ZnONPs characterization showed that they have a size ranging between 5.77 and 13.9 nm with a semi-sphere shape that is coated with a protein-carbohydrate complex. An EDX analysis of the Pm/ZnONPs revealed the presence of the shield matrix, which was composed of carbon, nitrogen, oxygen, chlorine, potassium, sodium, aluminum, sulfur, and zinc. The results of the FTIR analysis showed that the reduction and stabilization of the zinc salt solution were caused by the presence of O-H alcohols and phenols, O=C=O stretching of carbon dioxide, N=C=S stretching of isothiocyanate, and N-H bending of amine functional groups. The produced ZnONPs had good stability with a charge of - 16.2 mV, as evidenced by zeta potential analysis. The MTT assay revealed IC50 values of 8.42% and 200%, respectively, for the human A375 skin melanoma and human bone marrow 2M-302 cell lines. These findings revealed that the obtained Pm/ZnONPs have the biocompatibility to be applied in the pharmaceutical and biomedical sectors.

Zinc solubilizing bacteria and their potential as bioinoculant for growth promotion of green soybean (Glycine max L. Merr.).

Zinc-solubilizing rhizobacteria can convert insoluble zinc to an accessible form and increase Zn bioavailability in soil, which help mitigate Zn deficiency in crops. In this work, 121 bacterial isolates were isolated from the rhizosphere soils of peanuts, sweet potatoes, and cassava, and their capability to solubilize Zn was evaluated using Bunt and Rovira's agar containing 0.1% ZnO and ZnCO3. Among these isolates, six showed high Zn solubilization efficiencies ranging from 1.32 to 2.84 and 1.93 to 2.27 on the medium supplemented with 0.1% ZnO and ZnCO3, respectively. In a quantitative analysis of soluble Zn in liquid medium supplemented with 0.1% ZnO, the isolate KAH109 showed the maximum soluble zinc concentration of 62.89 mg L-1. Among the six isolates, the isolate KAH109 also produced the most indole-3-acetic acid (IAA) at 33.44 mg L-1, whereas the isolate KEX505 also produced IAA at 17.24 mg L-1 along with showing zinc and potassium solubilization activity. These strains were identified as Priestia megaterium KAH109 and Priestia aryabhattai KEX505 based on 16S rDNA sequence analysis. In a greenhouse experiment conducted in Nakhon Pathom, Thailand the ability of P. megaterium KAH109 and P. aryabhattai KEX505 to stimulate the growth and production of green soybeans was examined. The results revealed that inoculation with P. megaterium KAH109 and P. aryabhattai KEX505 considerably increased plant dry weight by 26.96% and 8.79%, respectively, and the number of grains per plant by 48.97% and 35.29% when compared to those of the uninoculated control. According to these results, both strains can be considered as a potential zinc solubilizing bioinoculant to promote the growth and production yield of green soybeans.

The "beauty in the beast"-the multiple uses of Priestia megaterium in biotechnology.

Over 30 years, the Gram-positive bacterium Priestia megaterium (previously known as Bacillus megaterium) was systematically developed for biotechnological applications ranging from the production of small molecules like vitamin B12, over polymers like polyhydroxybutyrate (PHB) up to the in vivo and in vitro synthesis of multiple proteins and finally whole-cell applications. Here we describe the use of the natural vitamin B12 (cobalamin) producer P. megaterium for the elucidation of the biosynthetic pathway and the subsequent systematic knowledge-based development for production purposes. The formation of PHB, a natural product of P. megaterium and potential petro-plastic substitute, is covered and discussed. Further important biotechnological characteristics of P. megaterium for recombinant protein production including high protein secretion capacity and simple cultivation on value-added carbon sources are outlined. This includes the advanced system with almost 30 commercially available expression vectors for the intracellular and extracellular production of recombinant proteins at the g/L scale. We also revealed a novel P. megaterium transcription-translation system as a complementary and versatile biotechnological tool kit. As an impressive biotechnology application, the formation of various cytochrome P450 is also critically highlighted. Finally, whole cellular applications in plant protection are completing the overall picture of P. megaterium as a versatile giant cell factory. KEY POINTS: • The use of Priestia megaterium for the biosynthesis of small molecules and recombinant proteins through to whole-cell applications is reviewed. • P. megaterium can act as a promising alternative host in biotechnological production processes.