Study on bioactive molecules involved in extracellular biosynthesis of silver nanoparticles by Penicillium aculeatum Su1

Abstract

In this paper, the biosynthesis of high-stable and biocompatible silver nanoparticles (AgNPs) was implemented by employing cell-free filtrate of Penicillium aculeatum Su1. The compositions analysis of reducing biomolecules in reaction system before and after AgNPs synthesis suggested that proteins were mainly involved in the biosynthesis process of AgNPs. Polyacrylamide gel electrophoresis (SDS-PAGE) analysis displayed that two main protein bands with molecular weights ranging from 66.2 to 116 KDa and 35 to 45 KDa were capped on the surface of AgNPs. The further identification of these protein bands by liquid chromatography-mass spectrometry (LC-MS/MS) analysis indicated that actin as a major protein component was responsible for stabilization of prepared AgNPs. The activity of nitrate reductase secreted by P. aculeatum Su1 was 73.73 ± 3.89 μg/(g · h). Furthermore, the dialysis assay showed that small molecular components had significant impacts on yield and particle size of biosynthesized AgNPs. Reduced nicotinamide adenine dinucleotide or reduced nicotinamide adenine dinucleotide phosphate (NADH or NADPH)-dependent nitrate reductases and other types of reductases or non-enzymatic bioactive molecules (≥ 3.5 KDa) might simultaneously participate in the biosynthesis process of AgNPs mediated by P. aculeatum Su1.

Biosynthesis of Metal and Metal Oxide Nanoparticles Using Microbial Cultures: Mechanisms, Antimicrobial Activity and Applications to Cultural Heritage

“…However, the mechanism behind these phenomena is not yet fully understood. Notwithstanding, several studies using plant extracts and microorganisms’ cell-free supernatants have been performed in an attempt to identify the molecules responsible for the reduction and stabilization (capping agents) of these NPs [ 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 ]. In these studies, several techniques have been applied.…”

“…Chromatographic techniques, such as gas chromatography (GC) [ 29 ], high-performance liquid chromatography (HPLC) [ 32 ], ultra-high-performance liquid chromatography (UPLC) [ 34 ], liquid chromatography (LC) [ 40 ] and gel permeation chromatography (GPC) [ 41 ], have been employed to separate and identify the several compounds present in the reaction solutions before and after synthesis. The latter are usually followed by identification techniques, such as mass spectrometry (MS) [ 29 , 40 ] and high-resolution mass spectrometry (HRMS) [ 34 ]. One study also used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), aiming to identify the capping agents of MNPs [ 39 ].…”

“…One study also used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), aiming to identify the capping agents of MNPs [ 39 ]. Separation of proteins present in the media using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) has also been performed [ 40 ] to try to prove their role in the reaction, as well as to determine their molecular weight. Quantifications of several compounds in both the media and nanoparticle suspensions were performed to understand their role in the reactions, as well as their relative contribution to the synthesis—e.g., DTNB, DNSA, DTNP and Folin-phenol assays [ 40 , 41 ].…”

“…Separation of proteins present in the media using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) has also been performed [ 40 ] to try to prove their role in the reaction, as well as to determine their molecular weight. Quantifications of several compounds in both the media and nanoparticle suspensions were performed to understand their role in the reactions, as well as their relative contribution to the synthesis—e.g., DTNB, DNSA, DTNP and Folin-phenol assays [ 40 , 41 ]. Spectrophotometric analysis of the NPs plasmon peak associated with changes in the reaction media was generally performed [ 27 , 36 , 37 , 38 , 40 , 41 , 42 ] with the assumption that higher absorbance values are related to increases in concentration, and the shifts in the peak toward higher wavelengths (toward red) are due to the increase in the diameter of the nanoparticle [ 47 ].…”

The construction of a lymphoma cell-based, DC-targeted vaccine, and its application in lymphoma prevention and cure

“…Optimizations have been tried, such as genetically modifying tumor cells to produce cytokines (GM-CSF, TGF-β, and so on), which enhanced the stimulation of antitumor response or relieved the immunosuppressive effect [ 32 , 33 ]. In addition, biomaterials with good biocompatibility have been recently developed to delivery therapeutics [ [34] , [35] , [36] , [37] , [38] , [39] , [40] ], which can be flexibly designed to meet the modification needs of vaccines to enhance antitumor immune activity [ [41] , [42] , [43] ]. Unfortunately, these optimizations cannot solve the existing challenges substantially, and multi-treatments provide new directions of lymphomas’ therapy potentially.…”