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Dairy Industry is growing fast using the modern biotechnological techniques and tactis. In manufacture of dairy products, biotechnology has been used for many centuries (cultured milk products, cheeses, processed milk by products) either using starter cultures or enzymes for milk clotting, cheese ripening acceleration, fat, protein or lactose hydrolyzate production or antimicrobial purposes.



Dairy products are recognized as healthy natural products. Modern developments in the biotechnology have opened up new and stimulating possibilities in dairying for enabling the accessibility of milk and milk products within the attain of poor and cater to the needs of large population. Dairy industry, in particular, can enormously promote through biotechnological inventions which can not only develop the overall quality and safety of processed dairy foods but also improve their commercial values for local consumptions and exports. Since, the significant commitment of dairy industry is to offer top notch nutritious, refreshing and cheap dairy nourishments to the shoppers; it has gotten unsurprising to join biotechnological mediation at an alternate phase of milk generation and preparing.
dairy industry milk indusrty lab experiment
Modern Recombinant DNA technology has been more focused on producing species which are specific to the certain enzyme production by manipulating and directing their genes. Below are the lists of the Industrial enzymes used in Dairy Industry popular in the 21st century Enzyme market:





Enzyme


Function


Microorganisms

Acid proteinase Milk coagulation Aspergillus sp.
Neutral proteinase Faster cheese ripening, debittering Bacillus subtilis, A. oryzae
Lipase Faster cheese ripening, flavor customized cheese, Aspergillus niger, A. oryzae
Lactase (β-galactosidase) Lactose reduced milk and whey products A. niger, Escherichia coli, Kluyveromyces sp.
Aminopeptidase Faster cheese ripening, Flavoring and taste improvement Lactobacillus sp.
catalase Cheese processing Aspergillus niger

Acid Proteinase and Neutral Proteinase

Proteinases are enzymes used in the milk industry to catalyze/hydrolyze the milk protein (casein), which stabilizes micelle formation preventing coagulation. The enzymes are widely used in the dairy industry for cheese processing.
Neutral proteinase refers to a class of proteinases that can act catalysis in a neutral, weakly acidic, or weakly alkaline environment. Its optimal pH is between 6.0 and 7.5, and can catalyze the hydrolysis of peptide bonds of proteins, releasing amino acids or peptides; where as Acid proteinase refers to class of proteinases that can act in acidic environment, helping in milk coagulation. 

Lipase

Lipases are the enzymes which are naturally found in our small intestine which digests the fats and lipids. This enzyme finds application in various industries including food, biofuel, detergents and animal feed. It is also used in leather, textile and paper processing applications. In the food and beverage industry, lipases find major application in dairy, baking, fruit juice, beer and wine industries. 
Commercial lipases are mainly used for flavor development in dairy products and processing of other foods containing fat. They can improve the characteristic flavor of cheese by acting on the milk fats to produce free fatty acids after hydrolysis. Different types of cheese can be made by using lipases from various sources, e.g. Romano cheese using kid/lamb pre-gastric lipase, Camembert cheese using lipase from Penicillium camemberti and cheddar cheese using Aspergillus niger or A. oryzae. Lipase catalysis could improve the texture and softness of cheese. Lipases are also used as flavor development agents in butter and margarine, also to prolong the shelf life of various baking products.
enzymes used in the dairy steps along with list of enzymes

Lactase Enzyme

It is popularly known as Lactase (beta galactosidase are enzymes classified as hydrolases). The enzyme digest the lactose found in milk into glucose and galactose. Because of intestinal enzyme insufficiency, some individuals even a population, show lactose intolerance and difficulty in consuming milk and dairy products. Hence, low-lactose or lactose-free food aid programme is essential for lactose intolerant people to prevent severe tissue dehydration, diarrhea, and at times, even death.
Another advantage of lactase-treated (to produce lactose free milk) milk is the increased sweetness of the resultant milk, thereby avoiding the requirement for addition of sugars in the manufacture of flavored milk drinks. Manufacturers of ice cream, yoghurt and frozen desserts use lactase to improve scoop and creaminess, sweetness, and digestibility and to reduce sandiness due to crystallization of lactose in concentrated preparations. Also Cheese manufactured from hydrolyzed milk ripens more quickly than the cheese manufactured from normal milk.
A. niger, Escherichia coli, Kluyveromyces sp. not only produces various enzymes but it is one of the few species classified as GRAS (Generally Recognized as Safe) by the Food and Drug Administration (FDA). A. niger is used in the production of enzymes, its cell mass is used as a component in animal feed and its fermentation produces organic acids and other compounds of high economic value.

Amino Peptidase

The proteolytic system of lactic acid bacteria contribute to the development of flavor during the ripening of cheese through the generation of short peptides and free amino acids, which directly or indirectly act as flavor precursors. Newly isolated lactic acid bacteria (LAB) as well as those procured from culture collection centers were screened for the production of various substrate specific amino peptidases. Peptidases are the enzymes used to break the bitter peptides found in the cheese and milk during processing to enhance the flavor and taste, Also it is used in faster cheese ripening. 

Catalase


Catalase is an enzyme found in all living organisms exposed to oxygen. It is mainly a tetrameric protein found in the aerobic organisms. It catalyzes the decomposition of hydrogen peroxide to water and oxygen. It helps the cell from protecting themselves from oxidative damage from reactive oxygen species. Catalase is used in food and dairy industry for the food preservation. Catalase is applied in milk processing industry to eliminate peroxide from milk, to remove glucose from egg white in baking industry and in food wrappers to prevent oxidation and control perishability of food. This enzyme has limited use in cheese production (Removal of hydrogen peroxide from milk prior to cheese production).
Biocatalytic capability of microorganisms have been utilized for a considerable length of time to deliver bread, wine, vinegar and other normal items without understanding the biochemical basis of their ingredients. Microbial Enzymes have become an interest of modern growing industries due their widespread uses in industries and medicine based on their stability, catalytic activity and the ease of production and its optimization as compared to any plant and animal based enzymes.



enzymes production

The application of Enzyme in various industries (e.g. Food, Agriculture, Chemicals and Pharmaceuticals) is growing rapidly due to reduced processing time, low energy input, cost effectiveness, non-toxic and eco-friendly properties. These microbial enzymes are well capable of degrading toxic compounds of industrial and domestic waste (i.e.: phenol compounds, nutriles, amines etc.) either via conversion or degradation.
Enzymes are large macromolecules composed of polymers of amino acids connected by amide bonds, ranging from kilodalton (insulin) to megadalton (ribosome) in molecular mass. Catalytic site of these macromolecules is often buried deep within hydrophobic pockets, which determines the specificity for their substrate. This specificity of enzyme to catalyze reactions between one types of chemical compound over the other provides the basis of its classification and name. With the great advancement achieved in the area of biochemistry after 1940, a large number of enzymes were isolated and characterized, and therefore, it was necessary to regulate the enzyme nomenclature. Thus, International Union of Biochemistry and Molecular Biology (IUBMB) in consultation with International Union for Pure and Applied Chemistry (IUPAC) established an Enzyme Commission (EC) to be in charge of guiding the naming and systematic classification for enzymes. According to the type of reaction catalyzed, the enzyme commission has classified the enzymes into six main classes, mentioned in below table:




Class

Reactions

Enzymes

Oxidoreductases

Transfer of hydrogen or oxygen or electrons between molecules

Dehydrogenases, oxidases, oxygenases, peroxidases

Transferases

Transfer of groups of atoms from one molecule to another

Fructosyltransferases, transketolases, acyltransferases, transaminases

Hydrolases

Hydrolytic cleavage of bonds

Proteases, amylases, acylases, lipases, phosphatases, cutinases

Lyases

Non-hydrolytic cleavage by elimination or addition reactions

Pectate lyases, hydratases, dehydratases, decarboxylases, fumarase, argino succinase

Isomerases

Transfer of group from one position to another within one molecule

Isomerases, epimerases, racemases


Modern Recombinant DNA technology has been more focused on producing species which are specific to the certain enzyme production by manipulating and directing their genes. Below are the lists of the Industrial enzymes popular in the 21st century Enzyme market.


 


Industry


Enzyme


Function


Microorganisms





























































Beverage

Pectinase

Depectinization

Aspergillus oryzae, Penicillium funiculosum

Glucose oxidase

Oxygen removal from beer

Aspergillus niger

Cellulase

Fruit liquefaction

Aspergillus niger, Trichoderma atroviride

α-Amylase

Starch hydrolysis

Bacillus, Aspergillus

β-Amylase

Starch hydrolysis

Bacillus, Streptomyces, Rhizopus

β-Glucanase

Restrict haze formation

Bacillus subtilis, Aspergillus spp.

protease

Restrict haze formation

Aspergillus niger

Pullulanase

Starch saccharification

Bacillus sp., Klebsiella sp.

Naringinase

Debittering

Aspergillus niger

limoninase

Debittering

Aspergillus niger, A. oryzae

Aminopeptidases

Protein breakdown during mashing

Lactobacillus brevis, L. plantarum

Glucose oxidase

Oxygen removal from beer

Aspergillus niger

Cellulase

Fruit liquefaction

Aspergillus niger, Trichoderma atroviride

α-Amylase

Starch hydrolysis

Bacillus, Aspergillus

β-Amylase

Starch hydrolysis

Bacillus, Streptomyces, Rhizopus

β-Glucanase

Restrict haze formation

Bacillus subtilis, Aspergillus spp.

protease

Restrict haze formation

Aspergillus niger

Pullulanase

Starch saccharification

Bacillus sp., Klebsiella sp.

Naringinase

Debittering

Aspergillus niger

limoninase

Debittering

Aspergillus niger, A. oryzae

Aminopeptidases

Protein breakdown during mashing

Lactobacillus brevis, L. plantarum



Animal feed

Phytase

Hydrolyze phytic acid to release phosphorous

Aspergillus niger

Xylanase

Enhanced digestibility of starch

Aspergillus sp., Bacillus sp.

β-glucanase

Digestive aid

Aspergillus niger



Pulp and paper

Lipase

Pitch control

Candida Antarctica

Protease

Biofilm removal

Bacillus subtilis

Amylase

Deinking, drainage improvement

Bacillus licheniformis

Xylanase

Bleach boosting

Trichoderma reesei, Thermomyces lanuginosus, Aureobasidium pullulans

Laccase

Non-chlorine bleaching, delignification

Bacillus subtilis

Cellulase

Deinking, drainage improvement

Bacillus sp., Aspergillus niger



Polymer

Lipase

Polycondensation, ring-opening polymerization of lactones, carbonates

Candida Antarctica

Laccase

Polymerization of bisphenol A

Trametes hirsuta

Glucose oxidase

Polymerization of anilines

Aspergillus niger, Penicillium chrysogenum

Transglutaminase

Crosslinking of protein

Streptomyces mobaraensis

Tyrosinase

Polymerization of lignin and chitosan

Trichoderma reesei



Detergent

Amylase

Carbohydrate stain removal

Aspergillus sp., Bacillus subtilis

Lipase

Fat stain elimination

Aspergillus oryzae, A. flavus,

Protease

Protein stain removal

Aspergillus oryzae, Bacillus subtilis

Cellulase

Color clarification

Aspergillus niger, Bacillus sp.

Cutinase

Triglyceride removal

Fusarium solani f. pisi

Mannanase

Mannan spot removal

Bacillus sp.



Leather

Alkaline protease

Dehairing, bating

Alcaligenes faecalis

Neutral Protease

Dehairing, soaking

Aspergillus niger, A. flavus, Bacillus subtilis

Lipase

Degreasing

Aspergillus oryzae, A. flavus,

Amylase

Fiber splitting

Aspergillus sp., Bacillus subtilis



Cosmetics

Superoxide dismutase

Free radical scavenging, skin care

Corynebacterium
Glutamicum, Lactobacillus plantarum

Protease

Removal of dead skin

Aspergillus niger, A. flavus, Bacillus subtilis

Endoglycosidase

Teeth and gum tissue care

Mucor hiemalis

laccase

Hair dye

Bacillus subtilis, Trametes versicolor

lipase

Skin care

Aspergillus oryzae, A. flavus



Organic synthesis

Lipase

Synthesis of pharmaceuticals, polymers, biodiesels, biosurfactants

Aspergillus oryzae, A. flavus

Glycosyl tranferase

Synthesis of oligosaccharides

Bacillus sp.

Nitrile hydratase

Synthesis of acrylamide, butyramide, nicotinamide

Rhodococcus rhodochrous PA-34, Bacillus sp. APB-6

Glucose isomerase

Production of High fructose corn syrup

Corynebacterium sp., streptomyces murinus

Acyltransferase

Synthesis of hydroxamic acids

Bacillus sp. APB-6

Laccase

Production of textile dyes, cosmetic pigments, flavor agents, and pesticides

Trametes versicolor, Bacillus subtilis



Waste management

Amidase

Degradation of nitriles containing wastes

Rhodococcus erythropolis

Amylase

Bioremediation of vegetables wastes

B. licheniformis, Aspergillus sp.

Amyloglucosidase

Starch hydrolysis for bioremediation

Aspergillus niger

Lipase

Degradation of crude oil hydrocarbons

Aspergillus oryzae, Candida tropicalis

Nitrile hydratase

Degradation of nitriles containing wastes

Rhodococcus sp.

Protease

Bioremediation of keratinic wastes

Chrysosporium keratinophilum

Laccase

Degradation of waste containing olefin unit, polyurethane and phenolic compounds

Trametes versicolor

Cutinase

Degradation of plastics, Polycaprolactone

Fusarium solani f. pisi

Manganese peroxidase

Degradation of phenolic compounds

Phanerochaete chrysosporium, Coprinus cinereus

Lignin peroxidase

Degradation of phenolic compounds

Phanerochaete chrysosporium, Coprinus cinereus

Oxygenase

Degradation of halogenated contaminants

Pseudomonas sp., Rhodococcus sp.


 source & credit National Center For Biotechnology And National Health Institute



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