Nanobiology Lab, Biotechnology Division, Council of Scientific and Industrial Research-Institute of Himalayan Bioresource Technology, Palampur, H.P. 176061, India.
J Nanobiotechnology. 2011 Dec 7;9:56. doi: 10.1186/1477-3155-9-56.
Elucidation of molecular mechanism of silver nanoparticles (SNPs) biosynthesis is important to control its size, shape and monodispersity. The evaluation of molecular mechanism of biosynthesis of SNPs is of prime importance for the commercialization and methodology development for controlling the shape and size (uniform distribution) of SNPs. The unicellular algae Chlamydomonas reinhardtii was exploited as a model system to elucidate the role of cellular proteins in SNPs biosynthesis.
The C. reinhardtii cell free extract (in vitro) and in vivo cells mediated synthesis of silver nanoparticles reveals SNPs of size range 5 ± 1 to 15 ± 2 nm and 5 ± 1 to 35 ± 5 nm respectively. In vivo biosynthesized SNPs were localized in the peripheral cytoplasm and at one side of flagella root, the site of pathway of ATP transport and its synthesis related enzymes. This provides an evidence for the involvement of oxidoreductive proteins in biosynthesis and stabilization of SNPs. Alteration in size distribution and decrease of synthesis rate of SNPs in protein-depleted fractions confirmed the involvement of cellular proteins in SNPs biosynthesis. Spectroscopic and SDS-PAGE analysis indicate the association of various proteins on C. reinhardtii mediated in vivo and in vitro biosynthesized SNPs. We have identified various cellular proteins associated with biosynthesized (in vivo and in vitro) SNPs by using MALDI-MS-MS, like ATP synthase, superoxide dismutase, carbonic anhydrase, ferredoxin-NADP⁺ reductase, histone etc. However, these proteins were not associated on the incubation of pre-synthesized silver nanoparticles in vitro.
Present study provides the indication of involvement of molecular machinery and various cellular proteins in the biosynthesis of silver nanoparticles. In this report, the study is mainly focused towards understanding the role of diverse cellular protein in the synthesis and capping of silver nanoparticles using C. reinhardtii as a model system.
阐明银纳米粒子（SNPs）生物合成的分子机制对于控制其尺寸、形状和单分散性非常重要。评价 SNPs 生物合成的分子机制对于 SNP 形状和尺寸（均匀分布）的商业化和方法学发展至关重要。单细胞藻类莱茵衣藻被用作阐明细胞蛋白在 SNPs 生物合成中的作用的模型系统。
C. reinhardtii 无细胞提取物（体外）和体内细胞介导的银纳米粒子合成揭示了尺寸范围为 5±1 至 15±2nm 和 5±1 至 35±5nm 的 SNPs。体内生物合成的 SNPs 定位于外周细胞质和鞭毛根的一侧，鞭毛根是 ATP 运输途径及其合成相关酶的位置。这为氧化还原蛋白参与 SNPs 的生物合成和稳定提供了证据。在蛋白质耗尽部分中，SNPs 的尺寸分布改变和合成速率降低证实了细胞蛋白参与 SNPs 生物合成。光谱和 SDS-PAGE 分析表明，各种蛋白质与 C. reinhardtii 介导的体内和体外合成的 SNPs 有关。我们通过 MALDI-MS-MS 鉴定了与生物合成（体内和体外）的 SNPs 相关的各种细胞蛋白，如 ATP 合酶、超氧化物歧化酶、碳酸酐酶、铁氧还蛋白-NADP⁺还原酶、组蛋白等。然而，这些蛋白质在体外孵育预合成的银纳米粒子时没有与之相关。
本研究提供了分子机制和各种细胞蛋白参与银纳米粒子生物合成的证据。在本报告中，主要研究重点是使用莱茵衣藻作为模型系统，了解各种细胞蛋白在银纳米粒子的合成和盖帽中的作用。
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