Joan Christie W. (11320120002)

Ong. Nicky Alexander (11320120006)

Aristya Wismoyo (11320120017)

Danika Augusta S. (11320120019)

Overview : Enzyme

• Are naturally occurring bio-catalysts found in the living organisms

• Combine molecules to produce new products or break molecule into smaller parts

• Require specific conditions for efficient functioning like optimal temperature, pH,

concentration.

• Lower “activation energy” (amount of energy required for reaction to occur)

• Without enzymes, reactions would not occur fast enough to sustain life

• Will only bind with certain substrates. Active site is where chemical reaction

occurs

Overview : Sources of Enzymes

• Enzymes can be attained from animals (chimosin, pepsin, trypsin), plants (ficin, papain,

bromelain), or microorganisms (pectinases, glucoamylase, alpha amylase).

• The reason for a tendency to use microbial enzymes on industrial scale :

– plentiful supplied and producer can control all phases of the enzyme production

– commercial enzyme preparation is well standardized

– can often produce several different enzymes producer can operate the fermentation plant to shift

the production to one enzyme in detriment of the others coping with the market fluctuations

– presents high growing capability huge amounts of enzyme can be produced in a

short period of time

– can be genetically manipulated DNA recombinant technique to improve its enzyme

production capability

Overview : Enzyme Production

• Commercially valuable enzymes are usually produced through processes such as

extraction from adequate sources, filtration, centrifugation, precipitation, purification,

drying, stabilization, standardization, and packaging

• In biotechnology, all unit operations comprised between filtration and packaging are

generally said downstream processing

• Enzymes of animal and plant origin are produced through maceration of tissues,

organs, leaves, and fruits - followed by extraction with water or organic solvent

• Differently, microbial enzymes are attained from either prokaryotic

(bacteria) or eukaryotic (yeasts, fungi, mainly) cells cultured in a

liquid or semi-solid medium, carried out in a special reactor called

fermenter, being the process known as fermentation

Enzymes and Health Care Industries

• Health care (medical) industry as generally consisting of :

– hospital activities

– medical and dental practice activities

– other human health activities such as acupuncture and music therapy

– health care equipment and services; and

– pharmaceuticals, biotechnology and related life sciences.

• Development of medical applications for enzymes have been at least as extensive

as those for industrial applications, reflecting the magnitude of potential rewards.

• Variety of enzymes and their potential therapeutic applications are

considerable

• At present, the most successful applications are extracellular : purely

topical uses, removal toxic substances and treatment of life-threatening

disorders within the blood circulation.

Topical Medication Uses

• Enzymes is used in skin care products and especially used for exfoliation and

anti-inflammation.

• For exfoliation :

– Certain enzymes are effective exfoliants and can be gentler than other methods like scrubs

and microdermabrasion.

– The upper layer of your skin is mostly made of dead skin cells containing keratin protein.

– The enzymes work by specifically breaking down the keratin protein, resulting in smoother skin.

– Enzymes can also penetrate the skin to help cleanse pores and improve skin tone and texture.

– Look for fruit enzymes like papain (papaya), pineapple or blueberry.

Medical Application of Enzymes

• There are four different types of uses for enzymes in medical case :

– Analytical

– Manipulative

– Therapeutic

– Industrial

• With those uses, the treatment of pharmaceutical enzymes range from

detection to defection

• Used in supplements and medication

• There are both good and bad side-effect

Analytical Uses

• Enzymes used as detector :

– Enzymes can be used to detect and measure amounts of glucose in blood

– Amount of glucose in blood and urine is an indicator for diagnosis of diabetes

– Detected by using enzyme glucose oxidase

– Catalyzes reaction between glucose and oxygen to form gluconic acid

– Biosensor measures the produced gluconic acid and then indicated by a color change.

• Enzymes used to diagnose disorders :

– Enzymes of liver, kidney and other types leak into blood during related disorders.

– Thus the corresponding enzyme presence in high levels in blood than normal

range indicates the specific disorder.

Enzyme Use

Aldolase muscle disorders

Glutamate oxalo acetate transfrerase myocardial infarction

Iso-citrate dehydrogenase acute hepatitis

Acid phosphatase prostate cancer

Manipulative Uses

• The goal : make an enzyme that has improved properties to improve health

• Changes created through : exchange of amino acids, modifications in enzymes

structure, and adding short peptide domains (that stabilize the enzyme and target

the cell membrane or DNA).

• Most common genetic alterations are with the creation of enzymes; introducing

point mutations, exchanging protein domains, and changing the structure

• Enzymes can also be manipulated by :

– engineering fusion proteins

– enzyme hybrids

• recruit established functions and properties from existing enzymes and

incorporate them into the engineered enzyme.

• Creation of hybrid enzymes can expand the potential uses of natural enzymes.

• Enzymes, or fragments of enzymes, could potentially serve as building blocks for

proteins capable of catalyzing reactions not observed in nature

Therapeutic Uses

• Enzymes used to treat disorders :

– In some disorders like low blood pressure, head or spinal injuries, there are chances of

formation of blood clots due to stagnation of blood inside the blood vessels.

– These clots can be life threatening as they can get stuck in small vessels of brain and obstruct

blood flow.

– These clots can be removed only by dissolution by enzymes that can break them such as

Streptokinase or Urokinase.

• Enzymes used to assist metabolism :

– In old patients, digestive capacity is low due to low secretion of digestive enzymes

– Hence their digestive system cannot metabolize some food materials efficiently.

– In such cases they can experience malnutrition, constipation etc.

– To aid digestion, enzymes like Papain are administered orally after food for easier

digestion.

Therapeutic Uses

• Enzyme supplements for enzyme deficiencies :

– Many of us suffer from milk intolerance mainly due to a genetic lack of the correct enzyme

secretion.

– Milk intolerance usually relates to an inability to digest lactose (milk sugar).

– Prolactazyme contains lactase and other milk digesting enzymes treats lactose intolerance.

• Enzymes used to assist drug delivery :

– Some drugs need to penetrate deeper tissues for better action.

– For this some enzymes are used along with drugs in intra-muscular injection

forms to help proper penetration of tissues.

– One of such enzyme is Hyaluronidase (a natural human enzyme present in human

sperm to help sperm penetrate uterine tissue and fertilize with ova).

– The same enzyme is manufactured by rDNA technology and administered along with

drugs to enable efficient drug delivery to the target site.

Some Important Therapeutic Enzymes

Factors Severely Reduces Potential Utility of Enzymes as

Therapeutic Agents for Treatment of Metabolic Diseases

• They are too large to be distributed simply within the body's cells

– This is why enzymes have not yet been successful applied to large number of human genetic

diseases

– To overcome this problem : ex : enzymes covalently coupled to target-specific monoclonal

antibodies are being used to avoid non-specific side-reactions

• Their effective lifetime within circulation may be only a matter of minutes

– To overcome this problem : disguise using covalent modification, entrapment

of enzyme within artificial liposomes, synthetic microspheres, and red blood

cell ghosts

– Although these methods are efficacious at extending the circulatory lifetime of

enzymes, they often cause increased immunological response and additionally

may cause blood clots

Factors Severely Reduces Potential Utility of Enzymes as

Therapeutic Agents for Treatment of Metabolic Diseases

• Being generally foreign proteins to the body, they are antigenic and

can elicit an immune response which may cause severe and life-

threatening allergic reactions, particularly on continued use – To overcome this problem : disguising the enzyme as an apparently non-proteinaceous

molecule by covalent modification

– Asparaginase, modified by covalent attachment of polyethylene glycol, has been shown

to retain its anti-tumour effect whilst possessing no immunosgenicity

– Clearly the presence of toxins, pyrogens and other harmful materials within a

therapeutic enzyme preparation is totally forbidden

– Effectively, this encourages the use of animal enzymes, in spite of their

high cost, relative to those of microbial origin

Application of Enzyme in Cancer Treatment

• Asparaginase treatment of acute lymphocytic leukemia

• Tumor cells are deficient in aspartate-ammonia ligase activity, which restricts their ability

to synthesize the normally non-essential amino acid L-asparagine. Therefore, they are

forced to extract it from body fluids

• Action of the asparaginase doesn’t affect functioning of normal cells which are able to

synthesize enough for their own requirements, but reduce free exogenous concentration

and so induces a state of fatal starvation in susceptible tumor cells

• A 60% incidence of complete remission has been reported in a study of almost 6000

cases of acute lymphocytic leukaemia

• The enzyme is administered intravenously

• It is only effective in reducing asparagine levels within the bloodstream,

showing a half-life of about a day (in a dog)

• This half-life may be increased 20-fold by use of polyethylene glycol-modified

asparaginase

Industrial Uses – Drug Manufacture

• Immobilized enzymes are used in manufacture of many drugs and antibiotics

• This is possible as enzymes convert the pro-drug molecules to drugs or starting

material to drugs

• In production of Antibiotics:

– Immobilized “pencillinase” is used in converting Pencillin-G or V to 6-amino pencillinic acid.(6-

APA).

– From this 6-APA Ampicillin is produced using immobilized enzyme “pencillamidase.”

– Other examples include: Bacitracin, tyrosine, nikkomycin are manufactured using

suitable immobilized enzymes.

• In Production of Steroids:

– Prednisolone is produced from cortisone using immobilized enzyme obtained

from curcuria lanata.

Industrial Uses – Drug Manufacture

• To produce amino acids:

– Immobilised “L-aspartase-4-decarboxylase” enzyme helps in conversion of L-Aspartic acid to L-

Alanine an amino acid.

• In production of L-DOPA:

– L-DOPA is a drug used in treatment of Parkinsonism. This can be produced by

using immobilized enzyme “β-tyrosinase.”

• In production of : Co-enzyme-A (Vit-B5), Pro-insulin, interleukin-2,

prostaglandins, monoclonal anti-bodies.

• To produce medicinal plant derivatives like anthraquinone

(a glycoside to relieve constipation).

References

Bheem. 2014. 12 Applications of Enzymes in Medicine, Food, & Other Industries. Retrieved

from : http://www.rajaha.com/applications-enzymes-role/ (7 September 2014).

Chaplin, M. 2014. Enzyme Technology : Medical applications of enzymes. Retrieved from :

http://www1.lsbu.ac.uk/water/enztech/medical.html (7 September 2014).

Madison, Michelle, Nancy, Peter and Vivian. 2014. Applications of Enzymes in the

Pharmaceutical Industry. Retrieved from : http://www.loreescience.ca/site/SBI4U/bioch

emistry/Applications%20of%20Enzymes%20in%20the%20Pharmaceutical%20Industry

.pdf (7 September 2014).

Nixon, A. E., Ostermeier, M., & Benkovic, S. J. 1998. Hybrid enzymes: manipulating enzyme

design. Trends in Biotechnology, 16 : 258 - 264.

Vitolo, M. 2012. Industrial Uses of Enzymes. Retrieved from : http://www.eolss.net/sample-

chapters/c17/e6-58-05-12.pdf (7 September 2014).