The hydrolysis of alpha-glucosidase linkage is the mechanism of all Family GH13 enzymes. The substrate size determines the rate of hydrolysis or the carbohydrate size. AMD is given as a non-sex-linked autosomal recessive condition, where the excessive accumulation of glycogen builds up within the lysosome vacuoles in nearly all types of cells and all over the body.
It is the most serious glycogen storage disease that affects muscle tissue. AMD is also categorized into three separate types according to the age of onset of the symptoms in affected individuals.
Infantile which is Type a , childhood which is Type b , and adulthood which is Type c. The AMD type is defined by the gene mutation type, which was localized in 17q At the same time, the mutation type will determine the production level of acid maltase.
AMD is fatal, and type-a generally dies of heart failure before age one. Where enzymes are produced Enzyme Substrate End-products Where produced Salivary amylase Starch Maltose Salivary glands Protease Protein Amino acids Stomach, pancreas Lipase Lipids fats and oils Fatty acids and glycerol Pancreas Pancreatic amylase Starch Maltose Pancreas Maltase Maltose Glucose Small intestine Where digestion happens Proteases catalyse the breakdown of proteins into amino acids in the stomach and small intestine Lipases catalyse the breakdown of fats and oils into fatty acids and glycerol in the small intestine Amylase catalyses the breakdown of starch into maltose in the mouth and small intestine Maltase catalyses the breakdown of maltose into glucose in the small intestine.
Salivary amylase. Salivary glands. Amino acids. In this way, maltase helps the entire digestive system function smoothly. Is sucrose an enzyme? Sucrase is a digestive enzyme that catalyzes the hydrolysis of sucrose to its subunits fructose and glucose. One form, sucrase-isomaltase, is secreted in the small intestine on the brush border.
What do you mean by enzymes? Enzyme: Proteins that speeds up the rate of a chemical reaction in a living organism. An enzyme acts as catalyst for specific chemical reactions, converting a specific set of reactants called substrates into specific products. Without enzymes, life as we know it would not exist. Where is protease produced? The body produces protease in the pancreas, but the pancreas doesn't produce protease in a working condition.
Instead, the protease produced in the pancreas has to be activated by another enzyme found in the intestine. Only after it is activated by the other enzyme, can the protease go to work breaking down protein. What is the purpose of enzymes? Enzymes are biological molecules typically proteins that significantly speed up the rate of virtually all of the chemical reactions that take place within cells.
They are vital for life and serve a wide range of important functions in the body, such as aiding in digestion and metabolism. After enrichment, the salts were removed from protein by performing dialysis against the same buffer overnight. These dialyzed precipitates were then used for further characterization. The catalytic activity of maltase was measured using GOD-PAP method using glucose as a reference sugar and maltose as a substrate [ 20 , 21 ].
Different buffers were evaluated for selection of suitable buffer as well as pH for enzyme substrate reaction. These buffers include acetate pH 5. After selecting the buffer, pH of that buffer was varied from 5. The pH stability of maltase was analyzed by pre-incubating the enzyme without substrate in different pH environment 5. The enzyme containing aliquots were retrieved and catalytic activity was monitored after adjusting the pH of all aliquots to optimum in order to examine the stability profile of the enzyme for potential industrial use.
The buffers used for this experiment were acetate buffer pH Maltase was further characterized on the basis of its thermal profiling by keeping the enzyme at different temperatures ranging from After every After every 10 days, the catalytic activity of enzyme was measured according to the standard assay procedure. To observe the substrate specificity, partially purified maltase was allowed to react with different substrates including maltose, starch, sucrose and lactose at optimum pH and temperature.
The influence of substrate concentration was also studied by performing the maltase assay at various concentration of maltose ranging from 2. The solutions were prepared in three different concentrations 1. The percent relative activity was compared with the control enriched enzyme without exposure to any metal ions. Different concentrations 1. The effect of different organic solvents DMSO, ethanol, methanol, isopropanol, formaldehyde and chloroform on the catalytic performance of maltase was investigated by pre-incubating the enzyme with different concentrations 1.
The enzyme was mixed with the sample diluting buffer ratio and the protein with known marker was initially loaded in their respective wells. Reservoir buffer of pH: 8. After this, the gel was stained with coomassie brilliant blue stain and kept overnight with constant stirring. The gel was destained using destaining solution until dark bands were appeared with a clear background. The molecular weight was measured using gel documentation system.
Zymography was performed as described by Pan et al. After performing electrophoresis, the gel was washed with double deionized water thrice and incubated with 7.
Then zymogram was removed and washed repeatedly with same buffer and kept in 2, 3, 5-Triphenyltetrazolium chloride solution dissolved in 1. In the current study, the maltase from B. The pH of the enzyme-substrate reaction mixture is among the important parameters that cause significant impact on the catalytic activity and stability of enzyme.
Therefore in this study, different pH ranges were used from 5. It was observed that as the pH of the reaction mixture increased, the catalytic activity of maltase also increased and reach to its maximum at pH Further increase in pH beyond optimum decreased the catalytic activity while less activity observed at pH The pH of the surrounding environment have great impact on charges of these groups and as a result of this the structural configuration of the enzyme can be changed which either increases or decreases the catalytic activity.
McWethyt and Hartman [ 24 ] reported similar kind of observations when maltase from Bacillus brevis was evaluated for its pH profile. Further, it was also noticed that increase in pH beyond optimum value also have negative impact on the catalytic activity and about It has also been reported that most of the enzymes showed their catalytic activity at neutral pH. Any change in pH either in acidic or alkaline side causes loss in activity, structural stability and solubility of protein as a result of changes in the charge groups [ 25 ].
Cihan et al. A that is relatively similar to current results where pH 6. Effect of various pH and temperature on the catalytic activity a, b and stability c, d of maltase. Consequently, the stability of maltase at these pH values was also investigated by pre-incubating the enzyme with different pH buffers ranging from 5.
It was found that the maltase was stable at pH The pH stability curve can be compared with pH optimum curve in order to understand the reversible and irreversible effects of pH. The reversible effect is the protonation of the amino acids in the active center while charge alteration of structurally important amino acids mostly generate irreversible changes in the enzyme native structure [ 27 ].
The loss of catalytic activity of maltase under extreme pH values for example pH 5. The reaction temperature is another critical factor for evaluation of the enzyme activity. The temperature is responsible to increase the kinetic energy of enzyme which ultimately increases the collision frequency of enzyme and substrate.
The catalytic activity of enzyme increases as the temperature increases up to its optimum temperature, beyond this point, the structural conformational of protein is disturbed due to increased kinetic energy of the molecules at higher temperature which eventually declines the catalytic performance of enzyme.
These results are very much similar to the previously reported findings of optimum temperature for catalytic activity of extracellular maltase from Bacillus brevis [ 24 ]. The stability of enzyme in response to different temperatures represents the capability of an enzyme to tolerate harsh industrial conditions. The thermophilic enzymes are also one of the major requirements to proceed different bioprocesses at commercial scale [ 28 , 29 ].
It was observed that the enzyme lost its catalytic activity as time increased. The loss of maltase activity at high temperatures due to exposure for longer time might be due to the increase in kinetic energy of the molecules.
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