The steady-state kinetic parameters of pyridoxal 5’-phosphate-dependent
recombinant methionine γ -lyase from three pathogenic bacteria,
Clostridium tetani, Clostridium sporogenes,
and Porphyromonas gingivalis, were determined in β-
and γ-elimination reactions. The enzyme from C. sporogenes is
characterized by the highest catalytic efficiency in the γ-elimination reaction
of L-methionine. It was demonstrated that the enzyme from
these three sources exists as a tetramer. The N-terminal poly-histidine
fragment of three recombinant enzymes influences their catalytic activity and
facilitates the aggregation of monomers to yield dimeric forms under denaturing
conditions. The cytotoxicity of methionine γ-lyase from C.
sporogenes and C. tetani in comparison with
Citrobacter freundii was evaluated using K562, PC-3, LnCap,
MCF7, SKOV-3, and L5178y tumor cell lines. K562 (IC50=0.4–1.3 U/ml),
PC-3 (IC50=0.1–0.4 U/ml), and MCF7 (IC50=0.04–3.2 U/ml)
turned out to be the most sensitive cell lines.
The recombinant producer of Rhodospirillum rubrum L-asparaginase (RrA) was received and purification procedure of RrA was developed. It was shown that RrA has following biochemical and catalytic characteristics: K for L-asn 0,22 мM, pH optimum 9,2; temperature optimum 54°С; pI=5,1±0,3; L-gln activity seems to be low-to-negligible. К562, DU145 and MDA-MB-231 cellular lines displayed significant sensitivity towards the enzyme (IC50=1,80; 9,19 and 34,62 ME/ml, respectively. In comparison with L-asparaginases from E . coli II type (EcA) and Erwinia carotovora (EwA) cytotoxicity of RrA seems to be higher than EwA, but lower than EcA. 10-fold i.p. RrA administration (4000 ME/kg per day) in L5178y bearing mice showed Т/С=172%. The received results show that RrA belongs to I type cellular L-asparaginases with low L-gln activity and the high antiproliferative effect.
Single crystals of potassium iron hydrogen phosphate, KFe3(HPO4)2(H2PO4)6 · 4 H2O, were prepared hydrothermally by heating a mixture of Fe2O3, H3PO4 and K2CO3 with a small amount of water. It crystallizes monoclinic, space group C2/c (N° 15 Int. Tab.) with Z = 4 and a = 1701(2), b = 960.4(5), c = 1750(1) pm, β = 90.88(7)°.
The crystal structure was solved by using 1716 unique reflections F0 > 4σ(F0) with a final wR2 value of 0.126 (SHELXL‐93).
The main feature of the crystal structure are layers formed by PO4‐tetrahedra around the FeO6‐octahedra parallel to (001). K+ and H2O molecules connect these layers. Effective Coordination Numbers (ECoN), Mean Fictive Ionic Radii (MEFIR), Charge Distribution (CHARDI) and the Madelung Part of Lattice Energy (MAPLE) are calculated for the title compound. The existence of hydrogen bonds is confirmed by these calculations.
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