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Nanomaterials 2019, 9, 1042 17 of 20 power of nanoparticles, might be responsible for the potent anti-cancer activity of AgNPs produced in this study. Collectively, the data suggests that biogenic AgNPs derived in this study can potentially inhibit cancer cell growth. Although the initial results seem promising, further studies on the mode of action and selectivity of AgNPs are required before AgNPs can be proposed as a suitable addition in the armoury against cancer cells. 4. Conclusions The extracellular pigment of T. purpurogenus was successfully utilized as a reducing agent to bio-synthesize AgNPs. A low pigment concentration of 0.5 g/L was required, which provides scope for scalability of the synthesis process. The optimum precursor concentration and reduction time were found to be 6 mM and 24 h, respectively. Characterization of the nanoparticles revealed their size to be in range of 4 to 41 nm. Further exploration is needed to understand the process of AgNP synthesis. The FTIR spectra of the pigment in natural and alkaline conditions were compared to understand the groups involved in the reduction mechanism, and the anti-microbial and anti-cancer activities of the biofunctionalized AgNPs were evaluated. The anti-microbial activity was detected to be more significant against Gram-positive S. epidermidis, which was ascribed to the presence of functional groups coating the exterior surface of the nanoparticles. Further research can be conducted for various combinations of antibiotics and AgNPs to understand their synergistic potential in the face of rising antibiotic resistance. The strong activity of the biofunctionalized AgNPs against HepG2 cancer cell line was also attributed to the molecules coating the nanoparticle surface. The mode of action of biogenic AgNPs and the reasons behind selectivity are still unclear and can be studied in detail in the future. More research is required to understand the molecules and functional groups present in the pigment, and to unearth other probable applications of the pigment-mediated biosynthesized AgNPs. Supplementary Materials: The following are available online at http://www.mdpi.com/2079-4991/9/7/1042/s1, Figure S1: (a) Qualitative EPMA analysis of sample showing presence of Ag between 3 and 4 keV. (b) Comparison with coverslip background showing the presence of impurities, Table S1: Semi-quantitative analysis of the targeted sample area by EPMA, Figure S2: (I) Effect of pigment on HEK-293, (II) HepG2 and, (III) HeLa cell lines. (a) Cell survival after exposure to 500 μg/mL of pigment (n = 3), (b) Cell morphology without exposure to pigment, (c) Cell morphology after exposure to pigment. Author Contributions: S.B. and H.A. designed the research; H.A. supervised the research; S.B., T.K., and H.A. designed the experiments and S.B., D.G., and K.O. performed them; S.B., T.K. and H.A. analysed, interpreted the results.; S.B., T.K., and H.A. wrote the manuscript. All authors read and approved the final manuscript. Funding: This work in part was supported in part by JSPS KAKENHI Grant-in Aid for Scientific Research B (19H03086), the Sumitomo Electric Industries Group Corporate Social Responsibility Foundation, and Noda Institute for Scientific Research GRANT 2018 (grant to H.A.). Acknowledgments: S.B. was supported by a scholarship grant from the Japanese Ministry of Education, Culture, Sports, Science, and Technology. D.G. also studied on the Global Food Security Partnership Program, University of Tsukuba. The manuscript has been edited carefully by native-English-speaking professional editor from Editage, a division of Cactus Communications. Conflicts of Interest: The authors declare no conflict of interest. References 1. Chandran, S.P.; Chaudhary, M.; Pasricha, R.; Ahmad, A.; Sastry, M. Synthesis of gold nanotriangles and silver nanoparticles using Aloe vera plant extract. Biotechnol. Prog. 2006, 22, 577–583. [CrossRef] [PubMed] 2. Mohapatra, B.; Kuriakose, S.; Mohapatra, S. Rapid green synthesis of silver nanoparticles and nanorods using Piper nigrum extract. J. Alloy Compd. 2015, 637, 119–126. [CrossRef] 3. Ali, K.; Ahmed, B.; Dwivedi, S.; Saquib, Q.; Al-Khedhairy, A.A.; Musarrat, J. Microwave accelerated green synthesis of stable silver nanoparticles with Eucalyptus globulus leaf extract and their antibacterial and antibiofilm activity on clinical isolates. PLoS ONE 2015, 10, e0131178. [CrossRef] 4. Srinithya, B.; Kumar, V.V.; Vadivel, V.; Pemaiah, B.; Anthony, S.P.; Muthuraman, M.S. Synthesis of biofunctionalized AgNPs using medicinally important Sida cordifolia leaf extract for enhanced antioxidant and anticancer activities. Mater. Lett. 2016, 170, 101–104. [CrossRef]PDF Image | Biosynthesis of Silver Nanoparticles Talaromyces purpurogenus
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