A new, more gentle enzyme purification for yeast enolase was developed. A series of kinetic experiments was performed with yeast enolase where the concentration of Mg(II) is kept constant and at the Km' level; the addition of Mn(II), Zn(II), or Cu(II) gives a hyperbolic decrease in the enzyme activity. The final velocity of these mixed-metal systems is the same as the velocity obtained only with Mn(II), Zn(II), or Cu(II), respectively. The concentration of the second metal that gives half-maximal effect in the presence of Mg(II) is approximately the same as the apparent Km (Km') value measured for that cation alone. Direct binding of Mn(II) to apoenolase in the absence and presence of Mg(II) shows that Mn(II) and Mg(II) compete for the same metal site on enolase. In the presence of D-2-phosphoglycerate (PGA) and Mg(II), only a single cation site per monomer is occupied by Mn(II). Water proton relaxation rate (PRR) studies of enzyme-ligand complexes containing Mn(II) and Mn(II) in the presence of Mg(II) are consistent with Mn(II) binding at site I under both conditions. PRR titrations of ligands such as the substrate PGA or the inhibitors orthophosphate or fluoride to the enolase-Mn(II)-Mg(II) complex are similar to those obtained for the enolase-Mn(II) complex, also indicating that Mn(II) is at site I in the presence of Mg(II). High-resolution 1H and 31P NMR was used to determine the paramagnetic effect of enolase-bound Mn(II) on the relaxation rates of the nuclei of the competitive inhibitor phosphoglycolate. The distances between the bound Mn(II) and the nuclei were calculated.(ABSTRACT TRUNCATED AT 250 WORDS)

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Ginkgo trees are common street trees in Korea, and the large amounts of leaves that fall onto the streets annually need to be cleaned and treated. Therefore, fallen gingko leaves have been used as a raw material to produce biochar for the removal of heavy metals from solutions. Gingko-leaf-derived biochar was produced under various carbonization temperatures and times. This study evaluated the physicochemical properties and adsorption characteristics of gingko-leaf-derived biochar samples produced under different carbonization conditions regarding Pb(II) and Cu(II). The biochar samples that were produced at 800 °C for 90 and 120 min contained the highest oxygen- and nitrogen-substituted carbons, which might contribute to a high metal-adsorption rate. The intensity of the phosphate bond was increased with the increasing of the carbonization temperature up to 800 °C and after 90 min of carbonization. The Pb(II) and Cu(II) adsorption capacities were the highest when the gingko-leaf-derived biochar was produced at 800 °C, and the removal rates were 99.2% and 34.2%, respectively. The highest removal rate was achieved when the intensity of the phosphate functional group in the biochar was the highest. Therefore, the gingko-leaf-derived biochar produced at 800 °C for 90 min can be used as an effective bio-adsorbent in the removal of metals from solutions.

There have been contradictory viewpoints whether soil amendments immobilize or mobilize heavy metals. Therefore, this study evaluated the mobility and bioavailability of Pb, Cu, and Cd in contaminated soil (1218 mg Pb per kg, 63.2 mg Cu per kg, 2.8 mg Cd per kg) amended with peat moss (0.22, 0.43, and 1.29% carbon ratio) and peat moss-derived biochar (0.38, 0.75, and 2.26% carbon ratio) at 0.5, 1, 3% levels. The more peat moss added, the stronger both mobility and bioavailability of Pb, Cu, and Cd would be. In contrast, the addition of peat moss-derived biochar significantly reduced both mobility and bioavailability of heavy metals through the coordination of metal electrons to C[double bond, length as m-dash]C (π-electron) bonds and increased pH. Maximum immobilization was observed in 3% peat moss-derived biochar treatment after 10 days of incubation, which was measured at 97.8%, 100%, and 77.2% for Pb, Cu, and Cd, respectively. Since peat moss and peat moss-derived biochar showed conflicting effectiveness in mobility and bioavailability of heavy metals, soil amendments should be carefully applied to soils for remediation purposes.

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