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- W3148633051 abstract "Alcaligenes faecalis strain A 1501 isolated from strain A15 was used in this experiment. Its main physiological and biochemical properties were the same as those of strain A15. Starin A1501, however,could use more sorts of carbohydrates as sole carbon source. A. faecalis A1501 was grown in N-free and combined N(NH_4~+30 mmol/L) culture medium, respectively. The Fe protein from N_2-grown cells (Af2) and NH_4~+grown cells (Af~* 2) were purified by treating the cell-free crude extract with DNase and RNase followed by 56℃ heating for 5 min. After centrifugation, the supernatant was twice passed through DEAE-cellulose (DE-52) columns(Fig. 1,2) followed by passing twice through Sephacryl S-200 column (Fig. 3) for gel filtration. Both Af2 and Af~* 2 were shown to be homogeneous by SDS-PAGE (Fig. 4). Af2 was purified 52-fold with specific activity of 1540 nmol C_2H_4 mg~(-1) protein min~(-1). No activity was detected from Af~* 2 (Table 1). The amino acid composition, metal contents, molecular weight, subunit composition, UV-visible spectrum and the properties of binding with MgATP and MgATP of Af2 and Af~* 2 were compared. The results showed that their physical and biochemical properties were almost the same. The molecular weight in both Af2 and Af~* 2 was 64.5 kD (Fig. 5) and the molecules were composed of two homogeneous subunits with the same molecular weight of 32.5 kD (Fig. 4). The number of amino acid resedues was 537 in Af2 and 553 in Af~* 2 and tryptophane was absent from both proteins(Table 2). Both Af2 and Af~* 2 contain 4 Fe and 4 acid-labile S~(2-). Their UV-visible spectrum features were the same,with a broad absorbance between 380 nm and 650 nm and with a small shoulder at 410 nm after being exposed to oxygen for 10 min(Fig. 6). Fluorescence probe FMA showed that both Af2 and Af~* 2 bound with 2 MgATP or 2 MgADP per molecule (Table 3)." @default.
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- W3148633051 date "1992-01-01" @default.
- W3148633051 modified "2023-09-25" @default.
- W3148633051 title "Biosynthesis and Properties of Fe Protein of Nitrogenase from N_2-Grown and NH_4~+-Grown Alcaligenes faecalis" @default.
- W3148633051 hasPublicationYear "1992" @default.
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