The enzyme action of myosaccharine

The principle
in many organisms, such as animals, plants,
microorganisms
, the anaerobic decomposition of sugar is almost identical process. This experiment takes the process of
of
in animal muscles and tissues. The enzyme action of creacosal glycogen, i.e. myosaccharides under the condition of lack of oxygen, after a series of enzymatic reactions, and finally into lactic acid process.
in muscle tissue first phosphate, through hexaglucose phosphate, propylene glyphosate, glyphosate and a series of intermediate products, and finally produce lactic acid. The process can be combined into the following reactive :of myosaccharine is a way for sugars to supply energy to tissues. When the body suddenly needs a large amount of energy, and oxygen supply is insufficient (e.g. intense exercise), the enzyme action of glycogen can temporarily meet the needs of energy consumption. Under aerobic conditions, the enzyme action of glycosin in tissues is inhibited, while aerobic oxidation is the main route of sugar metabolism.of glycogen enzymes, muscle erosion or muscle extract is generally used. When using muscle erosion, it must be carried out under anaerobic conditions, while with muscle extraction, it can be carried out under aerobic conditions. Because the enzyme system that catalyticly dissociates is all present in muscle extract, the enzyme system that catalytics the breathing action (i.e., the tricelic acid circulation and oxidation of the respiratory chain) is concentrated in the mitochondrials.of saccharin or starch, which can be observed by the production of lactic acid. After removing the
protein
and sugar, lactic acid can be cothermally with sulfuric acid into acetaldehyde, which in turn reacts with parabens to produce violet-colored substances, which are identified according to the appearance of color.method is more sensitive, each milliliter solution containing 1-5 μg lactic acid is given a clear color reaction. If there are a large number of sugars and proteins and other impurities, then serious interference with the determination, so the experiment should try to remove these substances. In addition, the instruments used in the determination should be strictly cleaned.
reagents
and equipment I, reagents parabens: called parabens 1.5g, dissolved in 100mL 0.5% NaOH solution, with 1.5% solution. If the color of the parabens is darker, recrystrystry with acetone or a waterless ethanol. After a long placement time, needle-like crystallization will appear and should be used after shaking well.0.5% glycogen solution (or starch solution); 20% tacrychloroacetic acid solution; calcium hydroxide (powder); thick sulphuric acid; copper saturated sulfate solution; 1/15mol/L phosphate buffer (pH7.4)., the materialrabbit muscle.III, equipment
test tube
1.5×15cm (×8) and test tube holder; pipe pipe 5mL (×2), 2 mL (×1), 1mL (×2); dropper; tube 10mL (×4); glass rod;
the
water bath;
how toFirst, prepare muscle moths after killing the rabbit, blood, immediately cut back and leg muscles, in low temperature conditions with scissors as far as possible to cut the muscles into muscle erosion. Note that it should be prepared before use. , muscle molycose saccharintake 4 test tubes, after the number of each add fresh muscle molycches 0.5g. Tubes 1, 2 are sample tubes and tubes 3 and 4 are blank tubes. Add 20% tricloste 3mL to the 3,4 blank tube and use a glass rod to break up the muscles and stir well to precipitate the protein and terminate the enzyme's reaction.
then added 3mL phosphoric acid buffer and 1mL 0.5% glycogen solution (or 0.5% starch solution) to each of the four test tubes. Stir well with a glass stick, add a little liquid paraffin to isolate the air, and place 4 test tubes in a heated water bath at 37 degrees C at the same time to keep warm.1.5h, remove the test tube and immediately add 20% tricloste 3mL to tube 1,2 to mix well. The contents of each test tube will be
filtered
, discarded to precipitate. Take 5mL of the filter fluid of each sample, add it to the numbered test tube, then add 1mL of copper sulfate solution to each tube, mix well, add 0.5g calcium hydroxide powder, stir it well with a glass stick, place 30min, and stir the contents from time to time, so that the sugar precipitation is complete. Each sample is filtered separately and precipitated. 3, lactic acid determination take 4 clean,
cand
test tubes, number, each test tube added to the thick sulfuric acid 2mL, the test tube to the cold water bath, respectively, with small The dropper took 1 or 2 drops of the filter fluid from each sample and added it to the cooled solution of the above-mentioned sulfuric acid (note that the size of the dropper is as consistent as possible), followed by shaking the test tube to avoid local overheating of the solution in the test tube. After mixing the test tube evenly, put it in a boiling water bath to cook 5min, remove it and cool it down, then add 2 drops of paraben reagents, do not drop hydroxybiphenyl reagents to the test tube wall, mix the test tube contents, compare and record the color depth of each test tube solution, and explain them.
notes are (1) Parabens must be purified to make them white. (2) In lactic acid determination, the test tube must be clean and dry to prevent contamination and affect the results.