Protein and DNA Isolation from Aspergillus Niger as well as Ghost Cells Formation

Abstract

Recently, a protocol for ghost cells preparations was introduced. It was given the name sponge-like protocol: Procaryotes, eucaryotes and virus were turned to ghost cells using such protocol. In this study, with slight modifications, Aspergillus niger ghost cells were prepared using the same protocol. Both the Minimum Inhibitory Concentration (MIC) and the minimum growth concentration (MGC) values for H2O2, NaOH, NaHCO3 and SDS against A. niger were determined. Five different randomization experiments were conducted instead of the full Plackett–Burman design. During the ghost preparation steps, the released Protein and DNA were measured spectrophotometrically at 280nm and 260nm, respectively. The quality of the prepared ghost cells were evaluated during the preparation steps using light microscope. Transmission electron microscope was used for evaluating the final steps. Protein and DNA electrophoresis were conducted to evaluate the quality of the released protein and DNA after each randomization experiment. The data obtained prove correct evacuation of the fungal cells from their cytoplasmic content during the successive steps. The study not only introduces a protocol for preparing ghost cells from Aspergillus niger but also enables the isolation of both of protein and DNA. The idea, the concept and the tools used in this study could establish a more sensitive method for protein and DNA isolation using any of four utilized chemical compounds. This proposes the same concept of enzyme-induced cell lysis which is based on minimizing the effect of used chemicals or enzymes. The study recommended extending the benefit of the sponge-like protocol from being a protocol for ghost cells preparation to DNA and protein isolation technique using the same concept.