STUDY OF ENZYME

Anupriya KapoorAssistant Professor

Institute Of Pharmacy, CSJM university, Kanpur, India

Introduction

• It refer to a class of enzymes that specifically split orhydrolyze starch.

• α-amylases: the amylases found in the animal kingdom• β-amylases: the one encountered in plant kingdom• bacteria and fungi give rise to α-amylases, althuogh the

enzymes from these two sources are not identical.• bacterial amylases are relatively heat-resistant with an

optimum temperature of ~ 55°C, which is a definiteadvantage over the corresponding fungal amylaseswhen starch hydrolysis is to be performed at a highertemperature

• A group of enzymes that hydrolyse complexsugars (starch, sucrose) into simplesugars(glucose, fructose,etc)

Complex sugarAmylase

Simple sugar

Applications of Amylase• Used as sweetening agent in food industry• Used in production of Dextrin• Used in production of Maltose syrup• Used to convert glucose to fructose• Used as starch sizing agent• Used in dry cleaning industry along with protease

enzyme.• Used in Brewing industry• Used in production of choclate syrup and corn

syrup

α-Amylases [Diastase]• It is an amylolytic enzyme or a mixture of enzymes.• obtained from1. fungi, for instance : Aspergillus oryzae2. nonpathogenic variant of bacteria, such as : Bacillus

subtilis3. α-Amylases, chemically known as [1, 4-α-

glucanglucanohydrolases]4. basically the extracellular enzymes that specifically

hydrolyze α-1, 4-glycosidic bonds5. these enzymes are known as endoenzymes, responsible

for affecting the cleavage of the substrate strategically

Study of enzymes: Hyaluronidase

Introduction • Hyaluronidases are a family of enzymes that catalyse

the degradation of hyaluronic acid (HA)• It is an enzyme found in the testes and semen• It depolymerizes hyaluronic acid, thereby enhancing

the permeability of connective tissues by dissolving thesubstances that essentially hold body cells together.Hyaluronidases may also be divided in 2 groupsaccording to pH-dependent activity:

• Acid active hyaluronidases: This group of enzymesexpresses their activity between pH 3 and 4.

• Neutral active hyaluronidases: active between pH 5and 8.

• Hyaluronidase is a dispersion agent, which modifiesthe permeability of connective tissue through thehydrolysis of hyaluronic acid, a polysaccharide found inthe intercellular ground substance of connective tissue,and of certain specialized tissues, such as the umbilicalcord and vitreous humor.

• Hyaluronidase For Injection [Wydase(R)] : It isobtained as a sterile dry, soluble enzyme productobtained from the mammalian (bovine) testes andcapable of hydrolyzing mucopolysaccharides of thehyaluronic acid type.

Pictorial representation of Action of Hyaluronidase enzyme

Hyaluronic acid

Hyaluronidase

splitting the glucosaminidic bond between C-1 of the glucosamine moiety and C-4 of glucoronic acid.

Change in membrane permeability

Therapeutic applications(1) It serves as intercellular cement, that essentially

binds together the paren-chymal cells. Probablythe enzyme, hyaluronidase affords thehydrolysis of hyaluronic acid by splitting theglucosaminidic bond between C-1 of theglucosamine moiety and C-4 of glucoronic acid.

(2) The most prominent clinical usage ofhyaluronidase is to distinctly facilitate theadministration of fluids by the aid ofhypodermoelysis (is the subcutaneousadministration of fluids to the body).

• Hyaluronidase injection is a spreading substance. It isused with other medicines given under the skin toimprove their uptake by the body. This method of drugdelivery is only used when the drug cannot be given byinjection into a vein. Hyaluronidase injection is alsoused to increase absorption of fluids or medicines thatare injected into your skin. It also used to help improveabsorption of radioactive substances during aprocedure called subcutaneous urography.

• This medicine is to be given only by or under the directsupervision of a doctor.

• This product is available in the following dosage forms:Solution

Cautions

(a) It must not be used in infected areas becauseof the danger of spreading the infection.

(b) Its usage with local anaesthetics is notrecommended.

US Brand Name

• Amphadase• Hydase• Hylenex• Vitrase

Side effects

More common• Bleeding, blistering, burning, coldness,

discoloration of the skin, feeling of pressure,hives, infection, inflammation, itching, lumps,numbness, pain, rash, redness, scarring,soreness, stinging, swelling, tenderness,tingling, ulceration, or warmth at the injectionsite

Introduction

• Penicillinase is a bacterial enzyme thatinvariably inactivates most but not allpenicillins (it is of β lactamase category)

• It is an extracellular type enzyme producedadaptively by members of the coliform groupof bacteria, by most Bacillus species, andcertain strains of Staphylococcus

• It causes the inactivation of Penicillin andCephalosporins

Cleavage of Penicillin

The enzyme Penicillin carries out hydrolysis of Penicillin to Penicilloic acid

• Some bacteria produce theenzyme penicillinase that destroys the beta-lactam ring of the antibiotic, making the penicillinineffective. Penicillinase resistantpenicillins are used to treat resistant strains ofstaphylococci and other infections.

• Penicillinase obtained from B. subtilis and B.cereus represent the industrially producedenzymes which exert action to some extent in theremoval of penicillin via specific inactivation.

Study of enzyme: Protease

introduction• Proteases or proteolytic enzymes refer to a class

of enzymes that exclusively break down orhydrolyze, the peptide bonds [—CO—NH—]which join the array of amino acids in a protein

• Proteases are usually produced by number ofbacteria, for instance : species belonging toBacillus ; Pseudomonas ; Closstridium, Proteus,and Serrata.

• It is also obtained from fungi, such as : Aspergillusniger ; Aspergillus oryzae ; Aspergillus flavus ; andPenicillium roquefortii.

Commercial Applications of Proteases

(1) Primarily used in ‘detergent industry’.(2) Emloyed in the ‘daily industry’, .e.g., rennin.(3)used in ’ in the ‘leather industry’,(4) Proteases also find their usage in the textile industry to afford proteinaceous

sizing.(5) In the ‘silk-industry’ proteases help in the liberation of the silk fibres from the

naturally occurring protenaceous material wherein they are actually imbedded.(6) Proteases (e.g., papain) are also employed as a meat tenderizer.(7) As an active and vital component in most spot-remover preparations for removing

foodspots in the dry-cleaning industry.(8) Other areas in which proteases are employed include : food industry, brewing

industry, film industry, waste-disposal (processing) management, and manufactureof protein hydrolyzates.

(9) Pharmaceutical industry also do make use of proteases in the manufacture ofcertain highly specific products for helping the suffering mankind

Classification of Proteases

(i) Alkaline proteases, (ii) Neutral proteases, (iii) Acid proteases, (iv) Fungal proteases, (v) Bacterial proteases