Statistical optimization and characterization of monodisperse and stable biogenic gold nanoparticle synthesis using Streptomyces sp. M137-2.

Microbial synthesis of gold nanoparticles (AuNPs), which are used in various forms with different properties in medicine, as a renewable bioresource has become increasingly important in recent years. In this study, statistical optimization of stable and monodispersed AuNPs synthesis was performed using a cell-free fermentation broth of Streptomyces sp. M137-2 and AuNPs were characterized, and their cytotoxicity was determined. The three factors determined as pH, gold salt (HAuCl4) concentration, and incubation time, which are effective in the extracellular synthesis of biogenic AuNPs, were optimized by Central Composite Design (CCD) and then UV-Vis Spectroscopy, Dynamic Light Scattering (DLS), X-Ray Diffraction (XRD), Scanning Electron Microscope (SEM), Scanning Transmission Electron Microscope (STEM), size distribution, Fourier-Transform Infrared (FT-IR) Spectroscopy, X-Ray Photoelectron Spectrophotometer (XPS) and stability analyzes of AuNPs were carried out. Optimum values of the factors were determined as pH 8, 10- 3 M HAuCl4, and 72 h incubation using Response Surface Methodology (RSM). Almost spherical AuNPs with 20-25 nm protein corona on the surface, 40-50 nm in size, monodisperse, and highly stable form were synthesized. Biogenic AuNPs were confirmed from characteristic diffraction peaks in the XRD pattern, UV-vis peak centred at 541 nm. The FT-IR results confirmed the role of Streptomyces sp. M137-2 metabolites in the reduction and stabilization of AuNPs. The cytotoxicity results also showed that AuNPs obtained using Streptomyces sp. can be used safely in medicine. This is the first report to perform statistical optimization of size-dependent biogenic AuNPs synthesis using a microorganism.

Adjuvant action of needle-shaped BC microfibrils.

Bacterial cellulose (BC) is an unbranched biopolymer produced by microorganisms and composed of glucopyranose units linked by ß-1,4 bonds. This study investigates the adjuvant action of needle-shaped BC microfibrils (BCmFs) in vitro using bovine serum albumin (BSA) as a model antigen. BC produced by the static culture of Komagataibacter xylinus was then microparticled (1-5 µm) by acid hydrolysis and characterized using Dynamic Light Scattering and Scanning Electron Microscopy. Subsequently, Attenuated Total Reflectance-Fourier-Transform Infrared Spectroscopy, cytotoxicity, TNF-α (tumour necrosis factor-alpha) and IL-6 (interleukin-6) cytokine secretion, and cellular uptake of the BCmFs-BSA conjugate on the human monocyte cell line (U937) differentiated into macrophages were performed. The microfibrils were determined to be 1-5 µm in size, needle-shaped, with a zeta potential of - 32 mV. Their conjugation with the model antigen, BSA, was demonstrated by FTIR analysis. In the cytotoxicity assay, BCmFs-BSA in macrophage cells showed high viability (over 70%). Although the highest TNF-α cytokine level (113 pg/ml) was obtained with BCmFs-BSA (Bovine serum albumin) conjugate (500 µg/ml) and was statistically significant (p = 0.0001) compared to the positive control group (BSA-aluminium hydroxide), IL-6 cytokine levels were not statistically different from those in the control group as desired. It has been shown in macrophage-differentiated U937 cells that microbially synthesized BC in the form of needle-shaped microfibrils (BCmFs) has a high cellular uptake capacity and increases the immunogenicity of the antigen. These results demonstrate for the first time that BCmFs have the potential to serve as a vaccine adjuvant.

Improvement of microstructure of cementitious composites by microbially-induced calcite precipitation.

In this study, microstructural improvement of cementitious composites was achieved by bacterial CaCO3 precipitation using three bacterial species, namely Sporosarcina pasteurii, Bacillus cereus, and Actinobacteria sp. M135-3, respectively. The final product was comparatively investigated regarding the physical effects of urease activity of different cells on the mortar in the long term.Microstructural improvement was determined by evaluating the pore structure by determining the increase in strength, decrease in water absorption, and capillary water absorption rate of the cement mortars having different microorganism concentrations (106-109 bacteria/ml). These measurements were taken on bacteria-containing and control samples on the 2nd, 7th, 28th, and 56th days, respectively. In addition, calcite and vaterite as calcium carbonate polymorphs formed by the precipitation of calcium carbonate by three types of bacteria were identified by Scanning electron microscopy and energy dispersive X-ray spectroscopy (SEM/EDS), X-ray diffraction (XRD) and Thermogravimetric analysis - Differential scanning calorimetry (TGA-DSC) analyzes.The bacteria-containing mortar samples showed that bacterial species and concentrations directly affect cementitious composites' mechanical and physical properties. Composite samples containing bacteria resulted in statistically significant microstructural improvements measured by higher mechanical strength, lower water absorption value, and capillary water absorption rate compared to control samples, especially at early ages. However, the effect of microbial calcite formation diminishes at later ages, especially at 56-days, attributed to the bacteria cells losing their vitality and integrity and forming spaces inside the mortars.

A Modular Chain Bioreactor Design for Fungal Productions.

Plastic bag bioreactors are single-use bioreactors, frequently used in solid culture fermentation. This study developed plastic bag bioreactors with more effective aeration conditions and particular connection elements that yield sensors, environmental control, and modular connectivity. This bioreactor system integrates the bags in a chain that circulates air and moisture through filtered connections. Within the present scope, this study also aimed to reveal that cultures in different plastic bags can be produced without affecting each other. In this direction, biomass production in the modular chain bioreactor (MCB) system developed in this study was compared to traditional bag systems. In addition, contamination experiments were carried out between the bags in the system, and it was observed that the filters in the developed system did not affect the microorganisms in different bags.