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Strong aroma of coffee is a mood refill for many not leaving me. Coffee stands as one of the world’s popular beverages and is well known for that robust aroma. All that strikes to mind is caffeine once we think about coffee. Caffeine is a Central Nervous System (CNS) stimulant and coffee is well known source of caffeine that is needed for human body. Do you know that coffee we get in supermarket is actually the “roasted seeds” of coffee cherries? And the only reason we call them beans is their close resemblance to legumes and both are not really related to one another. The two so called “beans” are enclosed in the “fruit called cherry” that is borne on short pruned coffee shrubs. Flowering is a continuous process in these plants so we can find green fruits alongside of ripe fruits on waxy leaf covered branches. Beans are inside the medium sized pulp covered by layers of exo, meso and endocarp along with pectin layers.

seed extract of green coffee

Journey from cherry to hot mug

The bright deep red coloured cherries are hand-picked carefully from the coffee fields and dried to maintain a moisture content of 11%. By now the pulp and other outer layers are separated out and only beans are left for further process. Roasting is the process where green coffee beans turn into dark brown colour and beans gain that characteristic aroma. During roasting the beans are placed in a metal cylinder. Hot air ranging from 180- 250 is propelled and cylinder is allowed to move continuously so that all beans are evenly roasted and do not burn out. Roasting beans induces certain chemical reactions and releases different volatile aromatic compounds that are responsible for the rich flavour of coffee. Roasted beans are powdered and brewed for making coffee.

Can you imagine coffee in some other colour than brown?

In the menu of a coffee bar we come across a number of coffee drinks like Espresso, Latte, Cappuccino, Americano, Mocha etc. Most of them are all the espresso-based drinks differing in proportions of espresso, milk (or cream) and foam. Recent days we have another one too into the menu. Can you imagine coffee in another colour? Green coffee is the new member mostly preferred by fitness lovers. Green coffees are simple hot drink made from unroasted raw coffee beans. Because the beans are unroasted, green coffee carries a different herbal flavour making it more different from regular coffee.The green coffee bean powder is brewed in water and served hot after it is filtered. Since it is not prepared in milk or cream it is cholesterol and fat free.

Chlorogenic acid aiding in weight loss

While other coffees are popular across the globe since quite olden times, green coffee has just started its way in grab attention among fitness beverages. It was first launched as an effective weight loss supplement. However, there was quite longer debate about its effectiveness. Chlorogenic acid (CGA) is the active ingredient aiding weight loss. Chlorogenic acid though naturally present in coffee beans, most of it is lost during the roasting process. Since green coffee is un-roasted and raw beans chlorogenic acid is higher. So, it was launched as weight loss supplement basing on its high CGA content.

Does it really have that“miraculous” effect as claimed during its launch?

Green coffee was stated as a miraculous weight loss supplement during its launch into market.    There was big health controversy over its weight loss effects.After conducting mice and human trails, researchers learnt that weight loss effects of chlorogenic acid in mice on high fat diet were almost negligible and that those obese mice were likely to develop Type-2 diabetes.

Other studies of green coffee in human fed with low fat, healthy diet and regular physical exercise showed quite good results. However, researchers believe studies are still needed in order to believe Green coffee as a weight loss supplement. More of human based trials having large group data are needed before a perfect conclusion is drawn. 

Thermogenic action of green coffee

Basically, human beings are endotherms. They maintain steady internal temperatures around 37 for the internal enzymes to be functionally active. When its hot outside produces sweat to cool off while it shivers during fall of external temperatures. Shivering is special muscular exertion that kicks off chilling experience. In this way endotherms remain less dependent on external temperatures by producing their own heat.



The internal heat generation consumes energy and since it is heat production its termed as thermogenesis. Fitness experts believe thermogenesis as one good way of burning calories. Thermogenesis is again exercise induced and botanical thermogenesis (based on herbal diets). Exercise induced thermogenesis causes more energy expenditure, increased protein synthesis thereby increased muscle mass. Botanical thermogenic agents facilitate a greater calorie utilization and provide the body with more some other benefits too.

Ephedrine, p-synephrine, caffeine, chlorogenic acid, capsaicin, capsiate, green tea, forskolin, carotenoids, isoflavones and flavonoids are herbal based natural thermogenic agents.

One thing to be kept in mind is that thermogenic foods only guarantee an increase in calorie burning but the effect can only be long lasting if the calories burning by physical activity accompanies it. Afterall supplements can only add some more sweat to workouts. But sole dependence on supplements never ensures good weight loss results.

Though the thermogenic action of natural agents is slow the effects of their consumption are wider. Caffeine is rich in coffee beans and is well known CNS stimulant. Caffeine is known as competitive inhibitor for enzyme phosphodiesterase. So, it inhibits the breakdown of 3′5′-cAMP. cAMP is a well-known signalling molecule of CNS.

Caffeine has adenosine receptor blockage activity. Basically, adenosine suppresses lipolysis so the activity of caffeine in blocking the receptor has an effect as increased lipolysis.  This makes free fatty acids more available for digestion. 

Also, caffeine induces the activation of adenylate cyclase (enzyme for cAMP production). Cyclic AMP produces a series of pharmacological effects like vasodilation, smooth muscle contraction, diuresis, increase in blood pressure etc. All of these are energy consuming processes.

Chlorogenic acid biochemically as a fat burner

Though coffeeis world popular beverage, knowledge about non-caffeine constituents is little. Chlorogenic acid is one among many non-caffeine constituents having biochemical importance. Chlorogenic acid is naturally present in coffee beans. During the roasting process, chlorogenic acid is converted to some other forms and roasted coffee has low CGA levels.

Chlorogenic acid chemically falls under polyphenols and is an ester of caffeic acid with quinic acid. Chlorogenic acid is whole set of hydroxy cinnamic esters of quinic acid with caffeoyl-, feruloyl-, dicaffeoyl- and coumaroyl- groups. Of all the isomers in each alkyl subgroups 5-caffeoylquinic acid is most common isomer of chlorogenic acid and is present in green coffee beans. 

Chlorogenic acid reduces the fat absorption from diet and reduce the stored fat levels. Researchers studied this action of chlorogenic acid directly and also as a constituent of green coffee and it showed to reduce carbohydrate absorption which helps in maintaining low blood sugar levels.Chlorogenic acid has thermogenic effects and green coffee is a combination of chlorogenic acid along with antioxidants that increase fatty acid availability in blood. This prevents unnecessary fat accumulation. 

Chlorogenic acid is an inhibitor of enzyme pancreatic lipase(hydrolyses the lipids in digestive tract). The enzyme is responsible for breaking the dietary triglycerides into simpler monoglycerides and free fatty acids. Since chlorogenic acid inhibits the pancreatic lipase activity in breaking the lipids therefore the fatty acid absorption through digestive tract is low.


Biochemically, chlorogenic acid in green coffee induces body fat loss. It acts as a physiological activator of peroxisome proliferator activated receptor alpha (PPAR-α). PPAR-α is mainly involved with free fatty acid oxidation in skeletal muscle and brown adipose tissue. 

Caffeine in green coffee release the fatty acids from stored fats. Chlorogenic acid induces liver for extra secretion of bile for improved processing of fatty acids. This combined effect can aid in loss of stored fats. Fitness experts design a combination of thermogenic foods along with physical exercise for reducing the waist lines in physical fitness seekers.
Both these effects on metabolism might have been preventing weight gain when the subjects are on regular physical exercise.

However, depending only on supplements for weight loss is never a perfect solution. Following a perfect regimen in both diet and exercise aspects along with supplements can show better results. After all, “supplements” can only add the “extra elemental effects” when worked with necessities. There can be no short cuts for weight loss without perfect eye watch on diet and physical work out.
Apart from its positive effects on glucose and fat metabolism, chlorogenic acid is reported to have anti-inflammatory, anti-carcinogenic and anti-bacterial activities. Its anti-carcinogenic properties are credited for free radical scavenging activities.

Reactive oxygen species (ROS) are highly reactive metabolic products of oxidative steps in various cellular organelles. Mitochondrial electron transport chain, cellular NADPH oxidase, endoplasmic reticulum, activated phagocytes produce ROS. Under normal physiological conditions ROS function as signalling messengers. An excessive generation of ROS can have negative impact on the oxidative balance of cellular environment. ROS at greater levels pose damage to DNA, proteins and lipids. Neurodegenerative disorders like Alzheimer’s, Parkinson’s, Huntington’s disease, diabetes, several cancers and ageing are caused when ROS exceeds their permissible levels.

Chlorogenic acid acts as a metal chelator and helps in scavenging free radicals. It has an inhibitory action on lipid peroxidation and NADPH oxidase activity both of which have role in ROS reduction. However, these anti-carcinogenic activities of chlorogenic acid are confined to cellular studies. Comprehensive studies aiming on organ systems with both long term and short-term consumption are yet on the way. Chlorogenic acid as a constituent is still to be studied for better understanding as functional food.

However, no studies so far have proved a negative shade on chlorogenic acid. Its biochemical properties boosted its value.  So, Green coffee can be a good choice of hot beverage if someone wants a new and a different try from the regular roasted coffee. It is available as green coffee beans as well as extract and the hot beverage is prepared by brewing it.

Roasted and Green coffee are pretty different though borne on same branch

  • It tastes quite different and mild than roasted coffee and is more like a hot mug of herbal tea.
  • Its origins from plants make it more likely to have anti-oxidant as well as anti-inflammatory effects.
  • Caffeine is relatively low in green coffee beans compared to regular roasted coffee.
  • Pushing back the weight loss aspects, green coffee has gained people attention for its distinct taste. Apart from chlorogenic acid Green coffee is also a source of caffeine but has some herbal flavour. After all both “Green and Brown” are from same branch. But there is big journey for turning from green to brown. Roasting changes the biochemical table of coffee beans.

 Recommended dosage

Green coffee can be tried as fitness beverage as it has been proved that no possible harm is imposed on ingesting it. A maximum of 3 or 4 cups a day for a healthy adult can be recommended. Diabetics, pregnant women and persons with high blood pressure need physician consultation before deciding safer intake.

/Disclaimer: The information provided in this article is just for the information of General Health Supplements. Science Arena Does not claim any health benefits nor its authors in any form, all the data and information shown are based on the personal experience and reference health data taken from Health practitioner. [Not For Medicolegal Purpose]/



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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