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Most of the reactions in living organisms are catalyzed by protein molecules called enzymes. Enzymes can rightly be called the catalytic machinery of living systems. The real break straight through of enzymes occurred with the introduction of microbial proteases into washing powders. The first market bacterial Bacillus protease was marketed in 1959 and major detergent manufactures started to use it around 1965.
The market enzyme producers sell enzymes for a wide collection of applications. The estimated value of world market is presently about Us$ 2 billion. Detergents (37%), textiles (12%), starch (11%), baking (8%) and animal feed (6%) are the main industries, which use about 75% of industrially produced enzymes.
Enzyme classification
Presently more than 3000 distinct enzymes have been isolated and classified. The enzymes are classified into six major categories based on the nature of the chemical reaction they catalyze:
1. Oxidoreductases catalyze oxidation or discount of their substrates.
2. Transferases catalyze group transfer.
3. Hydrolases catalyze bond breakage with the expanding of water.
4. Lyases take off groups from their substrates.
5. Isomerases catalyze intramolecular rearrangements.
6. Ligases catalyze the joining of two molecules at the charge of chemical energy.
Only a miniature whole of all the known enzymes are commercially available . More than 75 % of market enzymes are hydrolases. Protein-degrading enzymes constitute about 40 % of all enzyme sales. More than fifty market market enzymes are available and their whole is expanding steadily.
Enzyme yield
Some enzymes still extracted from animal and plant tissues. Enzymes such as papain, bromelain and ficin and other speciallity enzymes like lipoxygenase are derived from plants and enzymes pepsin and rennin are derived from animal. Most of the enzymes are produced by microorganisms in submerged cultures in large reactors called fermentors. The enzyme yield process can be divided into following phases:
1. Option of an enzyme.
2. Option of yield strain.
3. Building of an overproducing stain by genetic engineering.
4. Optimization of culture medium and yield condition.
5. Optimization of saving process.
6. Formulation of a carport enzyme product.
Criteria used in the Option of an market enzyme consist of specificity, reaction rate, pH and climatic characteristic optima and stability, corollary of inhibitors and affinity to substrates. Enzymes used in the market applications must commonly tolerant against discrete heavy metals and have no need for cofactors.
Microbial yield strains
In choosing the yield strain any aspects have to be considered. Ideally the enzyme is secreted from the cell. Secondly, the yield host should have a Gras-status. Thirdly, the organism should be able to yield high whole of the desired enzyme in a inexpensive life time frame. Most of the industrially used microorganism have been genetically modified to overproduce the desired activity and not to yield undesired side activities.
Enzyme yield by microbial fermentation
Once the biological yield organism has been genetically engineered to overproduce the desired products, a yield process has to be developed. The optimization of a fermentation process includes media composition, cultivation type and process conditions. The large volume market enzymes are produced in 50 -500 m3 fermentors. The extracellular enzymes are often recovered after cell dismissal (by vacuum drum filtration, separators or microfiltration) by ultrafiltration.
Protein engineering
Often enzymes do not have the desired properties for an market application. One Option is find a better enzyme from nature. Another Option is to engineer a commercially available enzyme to be a better market catalyst. Another Option is to engineer a commercially available enzyme to be a better market catalyst. Two distinct methods are presently available: a random method called directed evaluation and a protein engineering method called rational design.
Enzyme technology
This field deals with how are the enzymes used and applied in practical processes. The simplest way is to use enzymes is to add them into a process stream where they catalyze the desired reaction and are slowly inactivated during the process. This happens in many bulk enzyme applications and the price of the enzymes must be low to take their use economical.
An alternative way to use enzymes is to immobilize them so that they can be reused. Enzyme can be immobilized by using ultra filtration membranes in the reactor system. The large enzyme molecule cannot pass straight through the membrane but the small molecular reaction products can. Many distinct laboratory methods for enzyme immobilization based on chemical reaction, entrapment, specific binding or absorption have been developed.
Large scale Enzyme applications
1] Detergents
Bacterial proteinases are still the most important detergent enzymes. Lipases decompose fats into more water-soluble compounds. Amylases are used in detergents to take off starch based stains.
2] Starch hydrolysis and fructose production
The use of starch degrading enzymes was the first large scale application of microbial enzymes in food industry. Generally two enzymes carry out conversion of starch to glucose: alpha-amylase and fungal enzymes. Fructose produced from sucrose as a starting material. Sucrose is split by invertase into glucose and fructose, fructose separated and crystallized.
3] Drinks
Enzymes have many applications in drink industry. Lactase splits milk-sugar lactose into glucose and galactose. This process is used for milk products that are consumed by lactose intolerant consumers. expanding of pectinase, xylanase and cellulase heighten the liberation of the juice from pulp. Similarly enzymes are widely used in wine production.
4] Textiles
The use of enzymes in textile manufactures is one of the most rapidly growing fields in market enzymology. The enzymes used in the textile field are amylases, catalase, and lactases which are used to take off the starch, degrade excess hydrogen peroxide, bleach textiles and degrade lignin.
5] Animal feed
Addition of xylanase to wheat-based broiler feed has increased the available metabolizable power 7-10% in discrete studies. Enzyme expanding reduces viscosity, which increases absorption of nutrients, liberates nutrients either by hydrolysis of non-degradable fibers or by liberating nutrients blocked by these fibers, and reduces the whole of faeces.
6] Baking
Alpha-amylases have been most widely studied in connection with improved bread potential and increased shelf life. Use of xylanases decreases the water absorption and thus reduces the whole of added water needed in baking. This leads to more carport dough. Proteinases can be added to heighten dough-handling properties; glucose oxidase has been used to replace chemical oxidants and lipases to expand gluten, which leads to more carport dough and better bread quality.
7] Pulp and Paper
The major application is the use of xylanases in pulp bleaching. This reduces considerably the need for chlorine based bleaching chemicals. In paper development amylase enzymes are used especially in modification of starch. Pitch is a sticky substance present Generally in softwoods. Pitch causes problems in paper machines and can be removed by lipases.
8] Leather
Leather manufactures uses proteolytic and lipolytic enzymes in leather processing. Enzymes are used to take off unwanted parts. In dehairing and dewooling phases bacterial proteases enzymes are used to assist the alkaline chemical process. This results in a more environmentally kindly process and improves the potential of the leather . Bacterial and fungal enzymes are used to make the leather soft and easier to dye.
9] Speciality enzymes
There are a large whole of specialty applications for enzymes. These consist of use of enzymes in analytical applications, flavour production, protein modification, and personal care products, Dna-technology and in fine chemical production.
10] Enzymes in analytics
Enzymes are widely used in the clinical analytical methodology. Contrary to bulk market enzymes these enzymes need to be free from side activities. This means that expound purification processes are needed.
An important improvement in analytical chemistry is biosensors. The most widely used application is a glucose biosensor keen glucose oxidase catalysed reaction.
Several market instruments are available which apply this principle for estimation of molecules like glucose, lactate, lactose, sucrose, ethanol, methanol, cholesterol and some amino acids.
11] Enzymes in personal care products
Personal care products are a relatively new area for enzymes. Proteinase and lipase containing enzyme solutions are used for sense lens cleaning. Hydrogen peroxide is used in disinfections of sense lenses. The residual hydrogen peroxide after disinfections can be removed by catalase enzyme. Some toothpaste contains glucoamylase and glucose oxidase. Enzymes are also studied for applications in skin and hair care products.
12] Enzymes in Dna-technology
Dna-technology is an important tool in enzyme industry. Most primary enzymes are produced by organisms, which have been genetically modified to overproduce the desired enzyme. The specific order of the organic bases in the chain of Dna constitutes the genetic language. Genetic engineering means reading and modifying this language. Enzymes are crucial tools in this process.
13] Enzymes in fine chemical production
In spite of some successes, market yield of chemicals by living cells using pathway engineering is still in many cases the best alternative to apply biocatalysis. Isolated enzymes have, however, been successfully used in fine chemical synthesis. Some of the most important examples are represented here.
13 A] Chirally pure amino acids and aspartame
Natural amino acids are commonly produced by microbial fermentation. Novel enzymatic resolution methods have been developed for the yield of L- as well as for D-amino acids. Aspartame, the oppressive non-calorie sweetener, is synthesized in non-aqueous conditions by thermolysin, a proteolytic enzyme.
13 B] Rare sugars
Recently enzymatic methods have been developed to fabricate approximately all D- and L-forms of uncomplicated sugars. Glucose isomerase is one of the important market enzymes used in fructose manufacturing.
13 C] Semisynthetic penicillins
Penicillin is produced by genetically modified strains of Penicillium strains. Most of the penicillin is converted by immobilised acylase enzyme to 6-aminopenicillanic acid, which serves as a backbone for many semisynthetic penicillins.
13 D] Lipase based reactions
In expanding to detergent applications lipases can be used in versatile chemical reactions since they are active in organic solvents. Lipases used in transesterification and also used for enantiomeric divorce of alcohols and detach racemic amine mixtures. Lipases have also been used to form aromatic and aliphatic polymers.
13 E] Enzymatic oligosaccharide synthesis
The chemical synthesis of oligosaccharides is a complicated multi-step effort. Biocatalytic syntheses with isolated enzymes like glycosyltransferases and glycosidases or engineered whole cells are marvelous alternatives to chemical methods. Oligosaccharides have found applications in cosmetics, medicines and as functional foods.
Future trends in market enzymology
Industrial enzyme market is growing steadily. The hypothesize for this lies in improved yield efficiency resulting in cheaper enzymes, in new application fields. Tailoring enzymes for specific applications will be a hereafter trend with continuously enhancing tools and comprehension of structure-function relationships and increased search for enzymes from exotic environments.
New technical tools to use enzymes as crystalline catalysts, potential to recycle cofactors, and engineering enzymes to function in discrete solvents with manifold activities are important technological developments, which will steadily create new applications.
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