Get to Know about Hydrolysis of Carbohydrates Experiment

Hydrolysis of carbohydrates experiment is an experiment in which a chemical reaction breaks down carbohydrates through the addition of water. The process of breaking starch molecules into sugars is called hydrolysis and this experiment aimed to investigate whether this process can happen spontaneously or by using specific enzymes.

This post will give you a complete account on the steps to perform the experiment, how to set up the conditions, and some possible hypothesis from what might have happened if there is hydrolysis and no enzymes are used. This article will also provide useful tips for troubleshooting any problems that may arise during the experiment.

Benefits of hydrolysis of carbohydrates experiment

1. It is useful in studying the process of hydrolysis if there are no enzymes involved. Many enzymatic reaction are difficult to find out the exact role of each individual enzyme when they are expressed in small quantities, while here you can see them clearly.

2. If nothing happened and no enzymes were used, you can check that it is not because of factors such as lack or shortage of water, acids or temperatures supplied.

3. If something happened but without enzymes, you can see the physical changes that happen which will be helpful in studies on binding, etc.

Reagents used in hydrolysis of carbohydrates

1. Hypotonic solution

This can be prepared by dissolving a number of grams of Glucose in one litre of water. We normally use deionized water from the lab system (water from the distillate column that has been purified as it is our filtered tap water) but tap water can work too. The concentration of glucose when dissolved in deionized water is 0.1 M, which means the concentration will be 100 mM (About 10 g/L)

2. Protein

The protein used was bovine serum albumin, an abundant protein that is commercially available and also commonly used in lab nanotechnology projects, as well as a good indicator for hydrolysis because it forms hydrogen bonds with water molecules. This protein can be dissolved in water by adding a few drops of acetic acid.

3. Aqueous solution of enzyme

The enzymes used were amylase, or specifically alpha-amylase, an exo-enzyme which is typically extracted from fruits in an attempt to make easy access to this enzyme. The digestive enzyme sucrase was also used

Hypothesis on hydrolysis of carbohydrates experiment:

When one mixes the bovine serum albumin and the hypotonic glucose solution together, it will start absorbing water molecules from the surrounding environment and thus creating more hydrogen bonds with more water molecules and making a complex (denaturing). The formation of the complex will result in a lowering of the pH of the solution, denaturing the protein.

The addition of a hydrophobic enzyme such as alpha-amylase (which breaks cellulose and starch down into more polysaccharides) with high affinity for hydrophobic proteins and an exo-enzyme (not membrane bound, so it is more easily freed in solution) will start hydrolysing some small parts of this complex by breaking up some hydrogen bonds, thus reducing its size. The enzyme will then start digesting some other parts of the complex. This means that even if it was not put in the hypotonic solution, it could break down some of the complex by using its own hydrophobic enzyme and exo-enzyme.

Thus, as expected, as more enzyme is introduced into the solution hydrolysis will increase. This experiment can be considered a confirmatory experiment to show that hydrolysis of carbohydrates does happen spontaneously due to high concentrations of proteins, proteins which can also bind with water molecules and form hydrogen bonds as well as enzymes. However, if all these reactions are happening at very high rates without any enzymes then they should have been observed in a simple experiment with no extra ingredients or equipment required. This is an experiment that has been traditionally done by many students and chemists, though they usually end up repeating the same experiment. This experiment is a slightly complicated one and there will be some issues to look out for.

Inconclusions, as hydrolysis of carbohydrates experiment is a relatively simple experiment, there will not be any serious issues that you may need to work out. One thing that I observed is that the solution looks dull and the color does not change even if the concentration of glucose or pH was increased, although in fact this is due to the formation of cellulose and starch molecules. With this and other factors like very high amounts of hydrogen bonds in glucose or pH might have been too acidic in this case, it is difficult to conclude on what actually happened in each case. The possibility still exists that hydrolysis was not taking place at all because there were no enzymes involved.