Potentized Mercuric chloride and Mercuric iodide enhance α-amylase activity in vitro

Abstract

Mercuric chloride 30c and Mercuric iodide 30c were prepared by successive dilution in 30 steps of 1:100 followed by sonication at 20 KHz for 30 s at each step. Both were prepared in two media: 90% ethanol and distilled water. Three preparations of Mercuric chloride 30 in water were used: 12-month old, 1-month old and 4-day old. The controls for the water and ethanol–water preparations were pure water 30c and 90% ethanol 30c, respectively. For the three water preparations there were three matched controls of water 30c of the same ages. Each potentized substance or its control was mixed with distilled water 1 : 100 before testing. Hydrolysis of starch by α-amylase was measured by the standard procedure after incubation for 15 min at 27°C.

Mercuric chloride 30c and Mercuric iodide 30c in both water and aqueous ethanol media, enhanced enzyme activity significantly, compared to their respective controls. Mercuric chloride 30c, prepared in water 12 months previously, produced no significant change in the enzyme activity compared to its control.

We hypothesize that the structure of the active molecule imprinted on water polymers during the process of dynamization. The specifically structured water interacts with the active sites of α-amylase, modifying its activity. Ethanol molecules have large non-polar part stabilizing the water structure and thus retaining activity for a longer time.

Keywords

α-amylase - starch - hydrolysis - Mercuric chloride - Mercuric iodide - Mercurius corrosivus ageing

• References

• 1 Boyd WE. Biochemical and biological evidance of the activity of high potencies. Br Hom J 1954; 44: 6-44.
• 2 Sukul A, Sinhababu SP, Sukul NC. Reduction of alcohol induced sleep time in albino mice by potentized Nux vomica prepared with 90% ethanol. Br Hom J 1999; 88: 58-61.
• 3 Bernfield P. In: Colowick S, Kaplan NO. (eds). Methods of Enzymology. 1955: 149
• 4 Nelson DL, Cox MM. Lehninger Principles of Biochemistry. New York: Macmillan Worth Publishers; 2000. pp. 253–254, 550.
• 5 Petit C, Belon P, Got R. effect of homeopathic dilutions on subcellular enzymatic activity. Hum Toxicol 1989; 8: 125-129.
• 6 Qian M, Haser R, Payan F. Structure and molecular model refinement of pig pancreatic ∝-amylase at 2. 1A resolution. J Mol Biol 1993; 231: 785-799.
• 7 MacGregor EA, Janecek S, Svensson B. Relationship of sequence and structure to specificity in the ∝-amylase family of enzymes. Biochem Biophys Acta 2001; 1546: 1-20.
• 8 Kaboto S, Ogawa T, Muramoto K, Oosthuizen V, Naude RJ. The amino acid sequence of pancreatic ∝-amylase from the ostrich, Struthio camelus. Comp Biochem Physiol Part B 2000; 127: 481-490.
• 9 Rarey M, Kramer B, Lengauer T. The particle concept: placing discrete water molecules during protein–ligand docking predictions. Proteins 1999; 34: 17-28.
• 10 Perez- Gill JP, Cruz A, Casals C. Solubility of surfactant proteins in organic solvent/water mixtures. Structural studies on SP-B and SP-C in aqueous organic solvents and lipids. Biochem Biophys Acta 1993; 1168: 261-270.
• 11 Leroy A, Jonas A. Native-like structure and self-association behaviour of apolopoprotein A-I in a water/n-propanol solution. Biochem Biophys Acta 1994; 1212: 285-294.
• 12 Nedjar-Arroume N, Castellano A, Guillochon D. Stabilizing effect of water/alcohol solvents towards autoxidation of glutaraldehyde-modified haemoglobin. Biotechnol Appl Biochem 1995; 21: 173-183.
• 13 Wang JM, Takeda A, Yang JT, Wu C-S C. Conformation of concanavalin A and its fragments in aqueous solution and organic solvent–water mixtures. J Prot Chem 1992; 11: 157-164.
• 14 Halling PJ. High affinity binding of water by proteins is similar in air and inorganic solvents. Biochem Biophys Acta 1990; 1040: 225-228.
• 15 Fisher S, Verma CS. Binding of buried structural water increases the flexibility of proteins. Proc Natl Acad Sci USA 1999; 96: 9613-9615.
• 16 Watanabe H, Iwata S. Molecular orbital studies of the structures and reactions of a singly charged calcium ion with water clusters, Ca+ (H2O)n. J Phys Chem A 1997; 101: 487-496.
• 17 Muller A, Saenger W. Studies on the inhibitory action of Mercury upon proteinase K. J Biol Chem 1993; 268: 26150-26154.
• 18 Da Silva M CM, de Sa M FG, Chrispeels MJ. Togawa RC, Neshich G. Analysis of structural and physico-chemical parameters involved in the specificity of binding between ∝-amylases and their inhibitors. Protein Engng 2000; 13: 167-177.