Ten new thiosemicarbazone derivatives, furan-2-carbaldehyde thiosemicarbazone (1), 3-methyl-furan-2-carbaldehyde thiosemicarbazone (2), 5-hydroxymethyl-furan-2-carbaldehyde thiosemicarbazone (3), 5-trifluoromethyl-furan-2-carbaldehyde thiosemicarbazone (4), 5-nitro-furan-2-carbaldehyde thiosemicarbazone (5), 5-phenyl-furan-2-carbaldehyde thiosemicarbazone (6), 5-(2-fluorophenyl)-furan-2-carbaldehyde thiosemicarbazone (7), 5-(4-methoxyphenyl)-furan-2-carbaldehyde thiosemicarbazone (8), 5-(1-naphthyl)-furan-2-carbaldehyde thiosemicarbazone (9), and 5-(1H-Pyrazol-5-yl)-furan-2-carbaldehyde thiosemicarbazone (10) were synthesized by condensing thiosemicarbazide with the respective furan-2-carbaldehyde in methanol. The prepared compounds were characterized by spectroscopic studies (FT-IR and NMR) and electrospray mass spectrometry. The molecular structures of 2, 6, 7, and 8 have also been determined by X-ray crystallography. Compounds 2, 6, and 7 crystallize in the E conformation about the N1-C6, N1-C11, and N1-C11 bonds, respectively, while 8 adopts the Z conformation about the N1-C12 bond with the presence of an intramolecular N2-HO2 hydrogen bond. All prepared thiosemicarbazone derivatives were evaluated for their in vitro antibacterial, antifungal, and antitumor activities against Staphylococcus aureus strains, Candida albicans/Candida tropicalis fungi, and seven human tumor cell lines (HuTu80, H460, DU145, M-14, HT-29, MCF-7, and LNCaP), respectively. The antioxidant activity was also studied by the DPPH assay. Compound 5 exhibited significant antibacterial activity against Staphylococcus aureus ATCC700699 (MIC=1μg/mL) compared to the nitrofurantoin and gentamicin reference drugs (MIC=125 and 10->100μg/mL, respectively). Compound 4 was ten times less active than amphotericin B (MIC=5μg/mL) against Candida albicans (ATCC90028 and ATCC10231), while 1 exhibited a moderate effect of scavenging of DPPH radical (IC₅₀=40.9μg/mL) in comparison to ascorbic acid reference compound (IC₅₀=22.0μg/mL). Among all the studied thiosemicarbazones, 5 showed a higher cytotoxic activity (IC₅₀=13.3627.73μ) in relation to the other tested compounds (IC₅₀=34.84>372.34μ) against all tested cell lines, except the LNCaP cell line, exhibiting its highest antiproliferative activity (IC₅₀=13.36μ) on the HuTu80cell line. Besides, 8 and 9 exhibited high antitumor activity (IC₅₀=13.31 and 7.69μ, respectively) against the LNCaP cells.

Aims: To isolate and characterize lignocellulase producing thermophilic bacteria from a Peruvian hot spring. Study Design: Combined sediment and water samples from the hot spring were subjected to direct plating, in situ baiting and ex situ enrichment. Endoglucanase and xylanase producing bacterial colonies were isolated and characterized. Place and Duration of Study: Samples were taken from the Huancarhuaz hot spring, Peru (8º56’31.86”S, 77º47’00.53”W) in August 2010 and processed during 2011-2013. Methodology: Samples were subjected to three isolation methods and bacterial colonies with different color, size and appearance, were isolated, purified by streaking several times and conserved in Tryptic Soy Agar slants at 4ºC. The agar staining method was used to isolate enzyme-producing strains which were then identified by 16S rRNA sequencing and further studied for endoglucanase and xylanase production. Results: From 19 bacterial isolates only eight were selected for further study as they showed clearing activities on both carboxymethyl cellulose and xylan agar plates. By using 16S rRNA gene phylogenetic analysis, seven isolates were identified as Bacillus licheniformis and one as Cohnella laeviribosi which was the best xylanase and endoglucanase producer. Maximum endoglucanase activity produced by C. laeviribosi EHB4 was obtained at pH 6.0 and at 60ºC and only 50% of its activity was lost at 90ºC for 1h indicating that this enzyme is particularly thermostable. Conclusion: This is the first report on the production of endoglucanase by C. laeviribosi. These findings indicate that Peruvian hot springs are good sources of thermophilic cellulase-producing bacteria and that C. laeviribosi EHB4 may contribute to the development of biomass bioconversion processes.

En el presente trabajo estudiamos el grado de tolerancia a metales pesados de hongos y bacterias aisladas de suelos con y sin rizósfera, con el propósito de conocer su potencial para aplicaciones en biorremediación. Las muestras procedían del pasivo minero de Santa Rosa de Jangas. Los hongos y bacterias aislados fueron identificados taxonómicamente mediante el análisis de la región ITS y 16S ADNr, respectivamente. El índice de tolerancia a metales pesados se calculó usando medio salino suplementado con 1 mM a plomo (II), cobre (II), niquel (II) o zinc (II), y 0.1 mM a plata (I), cromo (VI) o cadmio (II). En total se aislaron 23 hongos y 18 bacterias. Las cepas de hongos con mejores índices de tolerancia fueron: Fusarium temperatum CTLM05 (Pb+2), F. temperatum CTLM08 (Zn+2), F. oxysporum CTLM18 (Ni+2 y Cd+2), F. oxysporum CTLM12 (Ag+1), F. inflexumCTLM22 (Cu+2) y Penicillium vanluykii CTLM11 (Cr+6). Las cepas de bacterias con mayores índices de tolerancia fueron Bacillus licheniformis SSR18 (Cd+2, Ni+2 y Zn+2), B. subtilis SSR3 (Pb+2), Serratia sp. SSR15 (Cu+2), Serratia sp. SSR13 (Ag+1) y B. cereus SSR01 (Cr+6). También se encontró que los hongos mostraron mejores índices de tolerancia que las bacterias. Finalmente, los suelos del pasivo ambiental minero de Santa Rosa de Jangas poseen una microflora interesante, probablemente con mecanismos para su adaptación, crecimiento y desarrollo sobre metales pesados y pueden ser de utilidad para el desarrollo de procesos biotecnológicos y biorremediación.

The lignocellulose bioconversion process is an eco-friendly and green-economy alternative technology that allows the reduction of pollution and global warming, so it is necessary for thermophilic and thermostable hydrolytic enzymes from natural sources. This research aimed to isolate cellulolytic and xylanolytic microbial consortia from Huancarhuaz hot spring (Peru) from sludge or in situ baiting cultured with or without sugarcane bagasse. According to the hydrolytic activities consortium T4 from in situ baiting was selected. It was cultivated in submerged fermentation at 65 °C, pH 6.5 for eight days using LB supplemented with sugar cane bagasse (SCB), pine wood sawdust (PWS), CMC, xylan of birchwood, or micro granular cellulose. Crude extract of culture supplemented with SCB (T4B) showed better endoglucanase and xylanase activities with higher activities at 75 °C and pH 6. In these conditions, cellulase activity was kept up to 57% after 1 h of incubation, while xylanase activity was up to 63% after 72 h. Furthermore, this crude extract released reduced sugars from pretreated SCB and PWS. According to metagenomic analysis of 16S rDNA, Geobacillus was the predominant genus. It was found thermostable genes: a type of endoglucanase (GH5), an endo-xylanase (GH10), and alkali xylanase (GH10) previously reported in Geobacillus sp. strains. Finally, Huancarhuaz hot spring harbors a genetic microbial diversity for lignocellulosic waste bioconversion in high temperatures, and the T4B consortium will be a promising source of novel extreme condition stable enzymes for the saccharification process.