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INTRODUCTION: Application of nanoparticles has been extensively increased in last decades. Nanoparticles of noble metals such as gold, platinum and especially silver are widely applied in medical and pharmaceutical applications. Although, variety of physical and chemical methods has been developed for production of metal nanoparticles, because of destructive effects of them on environment, biosynthetic methods have been suggested as a novel alternative. Some bacteria and microalgae have different ranges of potentiality to uptake metal ions and produce nanoparticles during detoxification process. In the present work, we study the potential of three Lactobacilli and three algal species in production of AgNPs in different concentrations of silver nitrate. METHODS: Utilizing AAS, XRD and TEM methods, Nannochloropsis oculata, Dunaliella salina and Chlorella vulgaris as three algal species in addition to three Lactobacilli including L. acidophilus, L. casei, L. reuteri were monitored for production of silver nanoparticles. Three concentrations of AgNO3 (0.001, 0.002, 0.005 M) and two incubation times (24h and 48h) were included in this study. RESULTS: Our findings demonstrated that C. vulgaris, N. oculata and L. acidophilus have the potential of nanosilver production in a culture medium containing 0.001 M of AgNO3 within 24 hours. Also L. casei and L. reuteri species exhibited their potential for production of silver nanoparticles in 0.002 M concentration of AgNO3 in 24 hours. The size range of particles was approximately less than 15 nm. The uptake rate of silver in the five species was between 1.0 to 2.7 mg/g of dry weight. Nanoparticle production was not detected in other treatments and the algae Dunaliella. CONCLUSION: The biosynthesis of silver nanoparticles in all of three Lactobacilli and two algal species including N. oculata and C. vulgaris was confirmed.
RESUMO
INTRODUCTION: Papaver somniferum is the commercial source of morphine and codeine. The isolation of effective genes involved in the morphine biosynthesis of P. somniferum is very important in the production of specific metabolites achieved using metabolic engi-neering techniques. In this pathway, the key enzyme COR is involved in the conversion of codeinone to codeine and morphinone to morphine. METHODS: the gene encoding of this enzyme was isolated using primers designed on the base of gene sequence available on (NCBI) for P. somniferum. This gene correct size around (960 bp) was first subcloned into pTZ57RIT vector then cloned into expression vectors (pBI121) between BamHI and SacI sites to allow the expression of cor gene driven by the cauliflower mosaic virus 35S pro-moter. The result was confirmed through different molecular methods e.g. PCR and en-zyme digestion by BamHI and SacI. The recombinant plasmid was transformed into the E. coli strain DH5α using a freeze-thaw method. Having selected positive colones on selection medium, plasmid was extracted by miniprep method and recombinant plasmids were selected based on PCR and digestion. The construct was then mobilized in Agrobacterium tumefaciens C58/pGV3850 (KmR RifR). After gene transformation to P. somniferum plants, the agroinfiltration method was also used for transient expression of COR enzyme. RESULTS: evaluation results showed that morphine and codeine were detectable in the leaves of transgenic plants containing cor transgene and there was significant difference in the final production. After completing this experiment for three times, results showed that in 11 sets from 15 sets of leaves experiment tested, main alkaloids (codeine, morphine, papaverin, noscapine and thebaine) were detectable. CONCLUSION: Whereas no signal was detected in non-infiltrated control leaves or in leaves infiltrated with non-recombinant bacteria for morphine and codeine, others such as thebaine and papaverine were detectable.
