School of Biotechnology, National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
ACS Nano. 2012 Jul 24;6(7):6165-73. doi: 10.1021/nn301502s. Epub 2012 Jun 25.
In recent years, there has been significant progress in the biological synthesis of nanomaterials. However, the molecular mechanism of gold biomineralization in microorganisms of industrial relevance remains largely unexplored. Here we describe the biosynthesis mechanism of gold nanoparticles (AuNPs) in the fungus Rhizopus oryzae . Reduction of AuCl(4)(-) [Au(III)] to nanoparticulate Au(0) (AuNPs) occurs in both the cell wall and cytoplasmic region of R. oryzae . The average size of the as-synthesized AuNPs is ~15 nm. The biomineralization occurs through adsorption, initial reduction to Au(I), followed by complexation [Au(I) complexes], and final reduction to Au(0). Subtoxic concentrations (up to 130 μM) of AuCl(4)(-) in the growth medium increase growth of R. oryzae and induce two stress response proteins while simultaneously down-regulating two other proteins. The induction increases mycelial growth, protein yield, and AuNP biosynthesis. At higher Au(III) concentrations (>130 μM), both mycelial and protein yield decrease and damages to the cellular ultrastructure are observed, likely due to the toxic effect of Au(III). Protein profile analysis also confirms the gold toxicity on R. oryzae at high concentrations. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis shows that two proteins of 45 and 42 kDa participate in gold reduction, while an 80 kDa protein serves as a capping agent in AuNP biosynthesis.
近年来,在生物合成纳米材料方面取得了重大进展。然而,工业相关微生物中金的生物矿化的分子机制在很大程度上仍未得到探索。在这里,我们描述了真菌 Rhizopus oryzae 中金纳米颗粒(AuNPs)的生物合成机制。AuCl(4)(-) [Au(III)]在 Rhizopus oryzae 的细胞壁和细胞质区域均被还原为纳米级的 Au(0)(AuNPs)。合成的 AuNPs 的平均尺寸约为 15nm。生物矿化通过吸附、初始还原为 Au(I)、随后络合[Au(I)复合物]和最终还原为 Au(0)发生。在生长培养基中,亚毒性浓度(高达 130μM)的 AuCl(4)(-) 会增加 Rhizopus oryzae 的生长并诱导两种应激响应蛋白,同时下调另外两种蛋白质。这种诱导会增加菌丝体生长、蛋白质产量和 AuNP 的生物合成。在更高的 Au(III)浓度(>130μM)下,菌丝体和蛋白质产量都会下降,并且观察到细胞超微结构受损,这可能是由于 Au(III)的毒性作用。蛋白质谱分析也证实了高浓度金对 Rhizopus oryzae 的毒性。十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析表明,两种 45 和 42kDa 的蛋白质参与金的还原,而 80kDa 的蛋白质则在 AuNP 的生物合成中作为封端剂。