RESUMO
A Cucurbita phloem exudate lectin (CPL) from summer squash (Cucurbita pepo) fruits was isolated and its sugar-binding properties and biological activities were studied. The lectin was purified by affinity chromatography and the hemagglutination assay method was used to determine its pH, heat stability, metal-dependency and sugar specificity. Antimicrobial and anticancer activities were also studied by disc diffusion assays and in vivo and in vitro methods. The molecular weight of CPL was 30 ± 1 KDa and it was stable at different pH (5.0 to 9.0) and temperatures (30 to 60 °C). CPL recovered its hemagglutination activity in the presence of Ca2+. 4-nitrophenyl-α-D-glucopyranoside, lactose, rhamnose and N-acetyl-D-glucosamine strongly inhibited the activity. With an LC50 value of 265 µg/mL, CPL was moderately toxic and exhibited bacteriostatic, bactericidal and antibiofilm activities against different pathogenic bacteria. It also exhibited marked antifungal activity against Aspergillus niger and agglutinated A. flavus spores. In vivo antiproliferative activity against Ehrlich ascites carcinoma (EAC) cells in Swiss albino mice was observed when CPL exerted 36.44% and 66.66% growth inhibition at doses of 3.0 mg/kg/day and 6.0 mg/kg/day, respectively. A 12-day treatment by CPL could reverse their RBC and WBC counts as well as restore the hemoglobin percentage to normal levels. The MTT assay of CPL performed against human breast (MCF-7) and lung (A-549) cancer cell lines showed 29.53% and 18.30% of inhibitory activity at concentrations of 128 and 256 µg/mL, respectively.
Assuntos
Anti-Infecciosos , Cucurbita , Lectinas de Plantas , Cucurbita/química , Animais , Lectinas de Plantas/farmacologia , Lectinas de Plantas/química , Lectinas de Plantas/isolamento & purificação , Camundongos , Humanos , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Antineoplásicos/farmacologia , Antineoplásicos/química , Linhagem Celular Tumoral , Carcinoma de Ehrlich/tratamento farmacológico , Carcinoma de Ehrlich/patologiaRESUMO
A seed lectin from Manilkara zapota (MZSL) was purified using ammonium sulphate precipitation and affinity chromatography. Hemagglutination activity, neutral sugar content and physicochemical properties of the lectin were determined and toxicity was checked by brine shrimp toxicity assay. Antimicrobial, antioxidant as well as in vitro anticancer activities of MZSL were also evaluated. Our findings showed the molecular weight of MZSL to be 33.0 ± 1 kDa. Minimum hemagglutination concentration of the lectin was 15.625 µg/ml. With a neutral sugar content of 6.32 %, the lectin was fully active at a temperature range of 30-50 °C and pH 7.0-8.0 and it was mildly toxic with an LC50 value of 107.93 µg/ml. The lectin demonstrated bacteriostatic activity against gram-positive bacteria in contrast to gram-negative bacteria at a concentration of 31.25 µg/ml, agglutinated Staphylococcus aureus and Shigella dysenteriae and exerted fungistatic activity against Aspergillus niger. MZSL dose-dependently reduced the formation of biofilm by E. coli. DPPH assay confirmed its antioxidant activity with an IC50 value of 96.42 µg/ml. MZSL showed 21.64 % growth inhibition against Ehrlich ascites carcinoma (EAC) cells at 80 µg/ml whereas its antiproliferative potential against MCF-7 and A-549 cancer cell lines became evident with IC50 values of 70.66 µg/ml and 107.64 µg/ml, respectively.
RESUMO
MytiLec-1, the recombinant form of a mussel lectin from Mytillus galloprovincialis, was purified by affinity chromatography and showed the maximum hemagglutination activity at a temperature range of 10 °C to 40 °C and at pH 7.0 to 9.0. Denaturants like urea and acidic-guanidine inhibited its hemagglutination activity significantly. MytiLec-1 was found to be metal-independent though Ca2+ slightly increased the activity of chelated MytiLec-1. The lectin suppressed 65 % growth of Pseudomonas aeruginosa (ATCC 47085) at 200 µg/ml and reduced the formation of biofilm (15 % at 200 µg/ml). Comparing to Shigella sonnei (ATCC 29930), Shigella boydii (ATCC 231903) and Shigella dysenteriae (ATCC 238135), Bacillus cereus (ATCC 14579) was slightly more sensitive to MytiLec-1. At a concentration of 200 µg/disc and 100 µg/ml, MytiLec-1 prevented the growth of Aspergillus niger and agglutinated the spores of Aspergillus niger and Trichoderma reesei, respectively. Amino acid sequences, physicochemical properties and antimicrobial activities of MytiLec-1 were compared with three other lectins (CGL, MTL and MCL from Crenomytilus grayanus, Mytilus trossulas and Mytilus californianus, respectively) from the mytilectin family of bivalve mollusks. It reconfirms the function of these lectins to recognize pathogens and perform important roles in innate immune response of mussels.