Mon, 18 Dec 2017

Anticancer and antiviral potential of folic acid conjugated with silver and gold Nanoparticles produced by Thermoduric Bacillus alcalophilus subsp halodurans

Rawhia Arafa

Department of Botany and Microbiology, Faculty of Science, Al-Azhar University, Egypt


ABSTRACT

Fifteen bacterial isolates were purified from Wady El Natrun loam salty calcareous sand soil textures and subjected to screen for their ability to produce folic acid. Bacterial isolate number WN12 was highly producer  of folic acid (Vitamin 9) and it was selected tentatively to be identified physiologically and phylogeny. The strain grows at 15-50°C, pH 6.0-12.5 and at salinities of 3.0-27.5 % NaCl (w/v). It was Gram positive , translucent brownish smooth color colony, long rods; non spore forming, motile by peritrichous numerous flagella.WN12 isolate identify as Bacillus alcalophilus subsp halodurans .The TLC Rf values for crude folic acid was 0.7 by using solvent system n-butanol, glacial acetic acid, water, methanol (20:10:20:5 v/v). The UV analysis of the fractions to the purified bacterial folic acid peak at 270 nm .Also, TEM images estimated the average size nearly around 0.1µm. While HPLC chromatograms of free folic acid maximum absorption wavelength was selected at 280 nm .And the FTIR spectra show absorption bands for pure folic acid infrared spectrum showed several identical absorption bands including OH, band at 3500 cm−1, C-H, band at 2900 cm−1 C=O band at 1100 cm−1. Comparison of the two synthesis conjugates Nano gold (AuNPs) and Nano silver(AgNPs) with folic acid (FA) characterization were  done . Six treatment were tested for measuring  infected dose 50 (IC50)  on four cancer cell lines,. Cytotoxic effect of  folic acid alone and Ag NP showed a variable pattern of reactivity to cell lines arranged ion in the order of MCF7> HCT> CaCO2> HEPG2 (P<0.05). AuNPs have no effect on HEPG2 and have highly effect on hct , MCF72 and Caco2 cell lines. Also, combination with FA showed the same pattern of antagonism. Preparation of trivalent mix of Ag-Au-FA showed antagonistic reaction to cell lines and toxicity of mix to cell lines was arranged in the order of MCF7 > CacO2 >HCT >HEPG2 (P<0.05).Also conjugated treatment  FA+AGNPs, (FA+AUNPs  and of trivalent mix of Ag-Au-FA have highly effect only on breast cancer(MCF7) IC50 . Finally Folic acid toxicity with the least concentration showed between 0.130 mg /ml to 0.180 mg/ml significant  IC50 high affecting  on different cell line. Antiviral potential were tested .Conjugated FA+AGNPs, FA+AUNPs were tested against A virus(HAV) and Vesicular stomatitis virus (VSV). Au-FA sample and Ag- FA  sample materials could induce viral load depletion in the order of 21 % and 74% for HAV post cell treatment with two test materials respectively. In the mean time VSV virus infectivity titer was decreased recording depletion of infectivity titer in the order of 3.75(68%) and 4.13(75%)for AuNP- FA- and AgNP-FA respectively.

Keyword: Folic acid, Bacillus alcalophilus subsp. halodurans bacteria, antitumor's, Antivirus ,nanosilver  and nanogol


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