ABSTRACT
The fungus Aspergillus flavus MTCC 873, a non-toxigenic isolate demonstrated its capability to synthesize mycoferritin (MF) upon induction with iron in yeast extract sucrose (YES) medium. The molecular mass, yield, iron and carbohydrate contents of the MF were 440 kDa, 0.015 mg/g of wet mycelia, 0.8 and 30.4%, respectively. Native gel-electrophoresis revealed a band corresponding to dimeric form of equine spleen ferritin (ESF). Subunit analysis by SDS-PAGE revealed a single protein band with an apparent molecular mass of 24 kDa, suggesting similar sized subunits in the structure of apoferritin shell. Immunological cross-reactivity was observed with the anti-fish liver ferritin. Transmission electron microscopy (TEM) revealed an apparent particle size of 100 Å. N-terminal amino acid sequence of MF revealed a sequence of SLPLQDYA, which showed identities with other eukaryotic ferritin sequences. The spectral characteristics (UV/VIS, fluorescence and circular dichroic spectra) were similar to ESF. The fungus, unlike A. parasiticus 255 (non-toxigenic) was incapable of producing aflatoxins, when grown in YES media.
Subject(s)
Amino Acid Sequence , Animals , Aspergillus flavus/chemistry , Electrophoresis, Polyacrylamide Gel , Ferritins/chemistry , Ferritins/isolation & purification , Ferritins/metabolism , Fungal Proteins/chemistry , Fungal Proteins/isolation & purification , Fungal Proteins/metabolism , Humans , Sequence Alignment , Spectrum AnalysisABSTRACT
To investigate the effect of placental isoferritin (PLF) on mouse embryo development in vitro, mice 2-cell embryos were co-cultured with human first trimester decidual cells at different concentrations of PLF in vitro. The following changes of the above system were observed under an invert microscope and the number of embryos were recorded and the embryos were classified. The results showed there was no significant difference in the percentage of embryos development to 4-cell, 8-cell and morula (P>0.05). PLF at the doses of 10 and 100 U/mL significantly enhanced more embryos development to the blastocyst and hatching blastocyst (P0.05). It was concluded that PLF at the concentration of 10-100 U/mL had no significant effects on the early development of mice embryos, however, PLF could promote the growth, differentiation, and hatching of preimplantation blastocysts.
Subject(s)
Coculture Techniques , Decidua/cytology , Embryo, Mammalian/cytology , Embryo, Mammalian/drug effects , Embryo, Mammalian/embryology , Embryonic Development/drug effects , Ferritins/isolation & purification , Ferritins/pharmacology , Placenta/chemistry , Tissue Culture TechniquesABSTRACT
Se purificó ferritina esplénica y hepática mediante el empleo de un inmunoabsorbente de Sepharosa 4B activada con CNBr-IgG antiferritina humana. Se realizaron 5 experimentos de purificación utilizando diferentes concentraciones y volúmenes de los extractos. el material no absorbido especificamente al inmunoabsorbente y la fracción cconteniendo ferritina se les realizaron estudios de pureza y recuperación. En cada experimento se obtuvieron alrededor de 500 *g de ferritina por Ml de inmunoabsorbente, electroforéticamente pura y con un patrón de corrida similar al de la ferritina obtenida por precipitación