In Vitro Cytotoxicity and Hemolysis Effect of Poly-Gamma-Glutamic Acid Nano-Polymer Biosynthesized Using Some Isolates of Bacillus spp.

Abstract

Natural polymers have good biological, biocompatible properties and a low degradation rate, so they can be used in medical applications. This study demonstrates an optimized biosynthesis for poly-gamma-glutamic acid nano-polymer (ɣ-PGA NP) using three bacterial isolates encoded as B4, B5, and B6. The 16S rRNA gene sequence of the isolates showed 98.5, 99, and 99.8% similarity with Bacillus sp., with accession numbers MZ976778, MZ976779, and MZ956153, respectively. The optimal conditions of the biosynthesis were 35 °C, followed by incubation for 30 h, 7 pH, 0.7 O.D600 of bacterial broth, carbon sources of glutamic acid and glucose, and ammonium sulfate as the nitrogen source. Biosynthesized ɣ-PGA NPs were characterized using X-ray diffraction (XRD), Zeta sizer, Fourier transform infrared (FT-IR), transmission electron microscopy (TEM), and nuclear magnetic resonance (1HNMR). The TEM study showed the formation of oval-shaped ɣ-PGA NPs with sizes 31.75–32.69 nm, 18.63–21.66 nm, and 37.57–39.22 nm for B4, B5, and B6, respectively. XRD, FT-IR, and 1HNMR confirmed the purity of the polymer. The hemolysis rates of the biosynthesized ɣ-PGA NPs were below the acceptable value. Moreover, the cytotoxicity test conducted using an MTT assay on peripheral blood mononuclear cells (PBMCs) revealed no significant effect on the cell viability within a dose range of up to 512 μg/mL.