{"title":"Enzyme Involvement in the Biosynthesis of Selenium Nanoparticles by Geobacillus wiegelii Strain GWE1 Isolated from a Drying Oven","authors":"Daniela N. Correa-Llant\u00e9n, Sebasti\u00e1n A. Mu\u00f1oz-Ibacache, Mathilde Maire, Jenny M. Blamey","volume":90,"journal":"International Journal of Bioengineering and Life Sciences","pagesStart":637,"pagesEnd":642,"ISSN":"1307-6892","URL":"https:\/\/publications.waset.org\/pdf\/9999976","abstract":"<p>The biosynthesis of nanoparticles by microorganisms,<br \/>\r\non the contrary to chemical synthesis, is an environmentally-friendly<br \/>\r\nprocess which has low energy requirements. In this investigation, we<br \/>\r\nused the microorganism Geobacillus wiegelii, strain GWE1, an<br \/>\r\naerobic thermophile belonging to genus Geobacillus, isolated from a<br \/>\r\ndrying oven. This microorganism has the ability to reduce selenite<br \/>\r\nevidenced by the change of color from colorless to red in the culture.<br \/>\r\nElemental analysis and composition of the particles were verified<br \/>\r\nusing transmission electron microscopy and energy-dispersive X-ray<br \/>\r\nanalysis. The nanoparticles have a defined spherical shape and a<br \/>\r\nselenium elemental state. Previous experiments showed that the<br \/>\r\npresence of the whole microorganism for the reduction of selenite<br \/>\r\nwas not necessary. The results strongly suggested that an intracellular<br \/>\r\nNADPH\/NADH-dependent reductase mediates selenium<br \/>\r\nnanoparticles synthesis under aerobic conditions. The enzyme was<br \/>\r\npurified and identified by mass spectroscopy MALDI-TOF TOF<br \/>\r\ntechnique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase.<br \/>\r\nHistograms of nanoparticles sizes were obtained. Size distribution<br \/>\r\nranged from 40-160 nm, where 70% of nanoparticles have less than<br \/>\r\n100 nm in size. Spectroscopic analysis showed that the nanoparticles<br \/>\r\nare composed of elemental selenium. To analyse the effect of pH in<br \/>\r\nsize and morphology of nanoparticles, the synthesis of them was<br \/>\r\ncarried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For<br \/>\r\nthermostability studies samples were incubated at different<br \/>\r\ntemperatures (60, 80 and 100 &ordm;C) for 1 h and 3 h. The size of all<br \/>\r\nnanoparticles was less than 100 nm at pH 4.0; over 50% of<br \/>\r\nnanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over<br \/>\r\n90% of nanoparticles have less than 100 nm in size. At neutral pH<br \/>\r\n(7.0) nanoparticles reach a size around 120 nm and only 20% of them<br \/>\r\nwere less than 100 nm. When looking at temperature effect,<br \/>\r\nnanoparticles did not show a significant difference in size when they<br \/>\r\nwere incubated between 0 and 3 h at 60 &ordm;C. Meanwhile at 80 &deg;C the<br \/>\r\nnanoparticles suspension lost its homogeneity. A change in size was<br \/>\r\nobserved from 0 h of incubation at 80&ordm;C, observing a size range<br \/>\r\nbetween 40-160 nm, with 20% of them over 100 nm. Meanwhile<br \/>\r\nafter 3 h of incubation at size range changed to 60-180 nm with 50%<br \/>\r\nof them over 100 nm. At 100 &deg;C the nanoparticles aggregate forming<br \/>\r\nnanorod structures. In conclusion, these results indicate that is<br \/>\r\npossible to modulate size and shape of biologically synthesized<br \/>\r\nnanoparticles by modulating pH and temperature.<\/p>\r\n","references":"[1] C. Ip, \u201cSelenium and ER stress response: Implication and exploitation\r\nfor cancer therapy\u201d. 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