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IN 309: Building Biotechnology: Science, Business, Law and Ethics Assignment 1:

Name of protein: Lysozyme Introduction: Lysozyme is the enzymes which plays important role in protecting us from the bacterial infection. It attacks on the protective cell wall of the bacteria which is made of with carbohydrate and peptide chains. Lysozyme breaks this carbohydrate chains and destroy the construction of the cell wall. After that, bacteria burst because of their own internal osmotic pressure. Due to its antibacterial effects, it is an important member of the immune system. Lysozyme found in tears, saliva, kidney tissue, cervical mucus, blood, milk, egg white,and other areas that are susceptible to potential bacteria growth. Apart from antibacterial characteristics, it is also responsible for the initial digestion of starches in mouth. Hen eggs white is the main sources of lysozyme for commercial supplies. Lysozyme was first officially characterized and noticed by the Alexander Fleming. It cannot be useful as drugs because it has large size so it is difficult to travel between the cells. Fleming continued his research and five years later he found true antibiotic drug: penicillin. Size and Characteristics: Lysozyme occupies an ellipsoidal shape with dimensions of roughly 45 x 30 x 30 and possesses a complex folding structure. The primary structure of lysozyme is single polypeptide which contains 129 amino acids. It consists of alpha () helixes and contains a -sheet structure in various areas along the polypeptide chain. Visually lysozyme is the White, lyophilized powder. It has pH level approx. 4.5 to 4.9. What does it do.? Lysozyme is known for damaging bacterial cell walls by catalyzing the hydrolysis of 1,4-beta-linkages between N acetylmuramic acid (NAM) and N-acetyl-D-glucosamine (NAG) residues in peptidoglycan, and between N-acetyl-Dglucosamine residues in chitodextrins. By breaking the 1,4 glycosidic bonds that link the sugars of these polysaccharides, the structural integrity of the bacterial cell wall is compromised and the bacterium bursts under the high internal osmotic pressure. Because it is commonly found in the places where microorganisms are most likely to enter the body, lysozyme is one of the powerful first line defenses against bacterial infection. Structure and Mechanism Lysozyme mainly has carbon, oxygen, nitrogen and sulfur atoms. It has amino acids residues. Like all other proteins it also contains 3C and 5N terminal. Lysozyme protein has two major domains (alpha and beta), and it has one active site as shown in figure. The alpha domain of the molecule is build up primarily of alpha helices (magenta), while the beta domain contains the only beta sheets (gold) and few helices. The active site is located deep within the cleft between the two domains. The deep gap between the two domains forms a cradle in which the long polysaccharides are confined. First enzymes (lysozyme) and substrate (polysaccharide) associate and creates the enzyme substrate complex. Enzymes catalyzed the hydrolysis reaction and it cleaves the substrate in to product which are quickly release and allowing the enzyme to catalyzed the another reaction.

Date: 20/ 01/2014

Submitted By: Tanay Patel (10002038) .

The cleft in the enzymes hold six sugar residues of a polysaccharide. The hydrolysis reaction occurs between these residues. Sugar residues have their most stable 3D configuration. However after the polysaccharide enter the enzymes-substrate complex, the enzymes forces the sugar into the strange configuration. There are two amino acid within the enzymes facilitate the reaction. Glu 35 and Asp 52. Glu 35 donates the proton to the sugar on the right. Asp 52 attacks on the C1 carbon atoms of the sugar on the left. This attacks on the C1 carbon results in the transient covalent bond between sugar and the amino acids and hydrolysis of the sugar-sugar bond. The deprotoned Glu 35 polarized the water molecule and trying to get proton which is lost. This allows the water oxygen to attacks the C1 carbon and breaks the C1 Asp 52 bond. In this way two amino acid returns in to their original states and forming the enzymes products complex. After that the enzymes and products dissociate.

Figure 1: Enzyme structure with substrate

Application of Lysozyme: Since lysozyme has been well known for its antibacterial and antifungal characteristics, it has wide verity of application in biochemical, pharmaceutical industries and food processing industries. In molecular biology, lysozyme is often used in the alkaline-lysis procedure for extracting and isolating plasmid DNA. It is used extensively in the pharmaceutical field for destroying gram-positive bacteria. Lysozyme can be also used for preventing the food (like dairy products) from spoilage in food industries. References: