In the current study, two different strains of the green, freshwater microalga Chlamydomonas reinhardtii bioreduced Ag+ to silver nanoparticles (AgNPs), which have applications in biosensors, biomaterials, and therapeutic and diagnostic tools. The bioreduction takes place in cell cultures of C. reinhardtii at ambient temperature and atmospheric pressure, thus eliminating the need for specialized equipment, harmful reducing agents or the generation of toxic byproducts. In addition to the visual changes in the cell culture, the production of AgNPs was confirmed by the characteristic surface plasmon resonance (SPR) band in the range of 415–425 nm using UV-Vis spectrophotometry and further evolution of the SPR peaks were studied by comparing the peak intensity at maximum absorbance over time. X-ray diffraction (XRD) determined that the NPs were Ag0. Micrographs from transmission electron microscopy (TEM) revealed that 97 ± 2% AgNPs were <10 nm in diameter. Ag+ to AgNP conversion was determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES). The AgNPs were stable over time in the cell culture media, acetone, NaCl and reagent alcohol solutions. This was verified by a negligible change in the features of the SPR band after t > 300 days of storage at 4 °C.
Insertional mutations exhibiting high cell-culture density HCD phenotypes are enriched through continuous subcultures in Chlamydomonas reinhardtii
