TRANSFERASES

The transferases catalyze the transfer of groups from one substrate molecule (donor molecule) to another (acceptor molecule). The reaction is typified by the equation: AX+BA+BX where A is the donor and B is the acceptor.

TRANSFERASES
Examples include transacetylases, transphosphorylases. TGases, transglycosidases, transmethylases, and

Some of them like TGases, and various glucanotransferases (e.g., fructosyl transferase, cyclodextrin glycosyl transferase (CGTase), and amylomaltase), are used extensively in the food industry.

Fructosyl Transferase
Fructosyl transferases catalyze the synthesis of fructose oligomers, also known as fructose oligosaccharides (FOS). The enzymes have been found in plants,
Onions and asparagus

microorganisms,
Streptococcus mutans, Fusarium oxysporum, B. subtilis, Aspergillus spp, Penicillium spp, and Aureobasidium spp

Fructosyl Transferase
Fructo-oligosaccharides account for about 30% of the sweetness of sucrose, but are non-digestible in the gastrointestinal tracts of humans thus they serve as dietary fiber render the gut milieu more conducive for the growth and proliferation of beneficial intestinal microflora. they are applied as low calorie sweeteners and as functional food ingredients with health benefits FOS are also used as sugar substitutes in foods such as light jam products, ice cream, and confectionery.

Fructosyl Transferase
The use of FOS in these foods helps to reduce
the caloric intake contributes to a lowering of blood glucose levels related health issues with diabetes.

The enzyme from B. subtilis has been used to synthesize fructose disaccharides such as xylosucrose, galactosucrose, and 6-deoxysucrose for use as
antigens, in glycosylated proteins, or as sweeteners or bulking agents in food products

Cyclodextrin Glycosyl Transferase

CGTase is also known by names like cyclodextrin glucanotransferase and cyclodextrin glucosyltransferase. CGTases have been characterized from various genera of bacteria, including Bacillus, Klebsiella, Pseudomonas, Brevibacterium, Micrococus, and Clostridium

Cyclodextrin Glycosyl Transferase

CGTases are known to catalyze at least four different reactions; coupling, cyclization, disproportionation, hydrolysis The most important of these reactions is intramolecular cyclization, whereby linear polysaccharide chains in molecules such as amylose are cleaved, and the two ends of the resulting fragments joined together to produce cyclic dextrins or cyclodextrins or CDs for short.

Cyclodextrin Glycosyl Transferase

CDs with varying units of glucose that are linked together by (1"4) glycosidic bonds have been described, namely the 6 glucose CD (or-CD), the 7 glucose CD (or-CD), and the 8 glucose CD (or-CD). By virtue of their ringed structures, CDs are able to trap or encapsulate other molecules to form inclusion composites with altered physicochemical properties that have great potential for applications in the (functional) foods, cosmetics, and fine chemicals

Amylomaltase
The enzyme amylomaltase, also known as 4-glucanotransferase, catalyzes the transfer of glucans from one-1,4-glucan to another -1,4glucan or to glucose.

The amylomaltase enzyme has been found in microorganisms,

Clostridium butyricum, E. coli

in plants
potato and barley
The enzyme acts on both malto-oligosaccharides and amylose to form medium to highly polymerized cycloamyloses that are highly

Amylomaltase
The enzyme acts on both maltooligosaccharides and amylose to form medium to highly polymerized cycloamyloses that are highly water-soluble These products are also expected to be of use in the (functional) food, pharmaceuticals, and fine chemical industries.

Transglutaminase
TGases are transferase enzymes that catalyze the acyl transfer reactions between free amino groups furnished by NH2 groups (e.g., the-NH2group of lysine) of amino acids and proteins, and the -carboxyamide groups of glutamines (Motoki and Seguro 1998). The TGase-assisted acyl transfer results in the extensive formation of strong and resistant covalent bonds between biomolecules like proteins. The products formed invariably have altered properties in terms of mechanical strength and solubility in aqueous systems. This feature of the enzyme is exploited in commercial food processing to improve the texture of gluten-free breads for individuals suffering from celiac disease (Moore et al. 2004, 2006), for making imitation crab meat and fish balls, and for binding small pieces of meat into larger chunks of meat. The enzyme is also used as a binding agent to improve the physical qualities (e.g., texture, firmness, and elasticity) of protein-rich foods such as surimi or ham, and emulsified meat products, such as sausages and hot dogs. They are also used to improve the texture of low-grade meats such as the so-called pale, soft, exudative meat or PSE meats; to improve the consistency of dairy products (milk and yogurt) to make them thicker and creamier; and to make noodles firmer (Kim and Kim 2009). TGases have been found to be present in animals (e.g., threadfin bream, big-eye snapper, white croaker (Benjakul et al. 2010), guinea pig (Huang et al. 2008), in birds, amphibians, and invertebrates (Lantto et al. 2005), and in plants and microorganisms (e.g., Bacillus, Streptoverticullium,andStreptomycesspp (Yokoyama et al. 2004)

Proteases The term protease is used to represent the group of enzymes that catalyze the cleavage of peptide bonds in proteins and peptide molecules with the participation of water as coreactant. Other names for this group of enzymes include proteinases or proteolytic enzymes. The majority of industrial enzymes currently in use are proteases, and it is estimated that they constitute about 40% of the total global enzyme production and use (Layman 1986, Godfrey and West 1996). These enzymes are used in the food processing, animal feed, detergent, leather and textiles, photographic, and other industries (Kalisz 1988). Proteolytic enzymes are also used as a digestive aid to facilitate digestion and use of proteins by some people who otherwise cannot readily digest these molecules. On the basis of their mode of action on the protein/peptide molecules, proteases are classified as either endopeptidases (or endoproteases) or exopeptidases (or exoproteases). Endopeptidases are those proteases that cleave peptide bonds randomly within protein molecules to result in smaller peptides. Examples include trypsin, chymotrypsin, pepsin, chymosin, elastase, and thermolysin. Exopeptidases cleave peptide bonds by successively removing terminal amino acid residues to produce free amino acids and the residual polypeptide molecule. Thus, two types of exoproteases are distinguished, the carboxypeptidases, that is, those that cleave amino acids at the carboxyl or C-terminal of the protein or polypeptide molecule, and the

Acid Proteases
The acid proteases (also known as aspartate proteases) are so called because the enzymes in this group tend to have one or more side chain carboxyl groups from aspartic acid as essential component(s) of their active sites, and they are optimally active within the acid pH range. Examples include pepsins, chymosins, rennets, gastricins (from animal sources), cyprosin, and cardosin (from plant sources), and several other aspartic acid proteases from microorganisms such as Rhizomucor miehei, R. pusillus, R. racemosus, Cryphonectria parasitica, and A. niger(Mistry 2006). Most of the known acid proteases are inhibited by the hexapeptide pepstatin. The major food industry use of acid proteases is in dairy processing, such as for the milk coagulation step in cheese making. In cheese making, the acid proteases added (e.g., rennet of chymosin) act to cleave critical peptide bonds between phenylalanine and leucine residues in-casein of the milk to form micelles that aggregate and precipitate out as the curd. The characteristic specificity of these proteases in these dairy products is important to restrict hydrolysis to the peptide bonds between the critical phenylalanine and leucine residues in -casein; otherwise, excessive proteolysis could elicit undue texture softening, bitterness, and/or off-odors in the product(s).

Serine Proteases
The serine proteases are the most abundant group of the known proteases (Hedstrom 2002). The characteristic features of this group of enzymes include the presence of a seryl residue in their active sites, as well as a general susceptibility to inhibition by serpins, organophosphates (e.g., di-isopropylphosphofluoridate), aprotinin (also known as trasylol), and phenyl methyl sulfonyl fluoride. Examples of the animal serine proteases are chymotrypsin, trypsin, thrombin, and elastase; examples of plant serine proteases are cucumisin (from melon; Kaneda and Tominaga 1975), macluralisin, and taraxilisin (from the osage orange fruit and dandelion, respectively; Rudenskaya et al. 1995); and examples of microbial serine proteases are proteinase K and subtilisin A (Couto et al. 1993). Several of the serine proteases function optimally within neutral to alkaline pH range (pH 711), and they find extensive use in the food and animal feed industries. They are used to produce highly nutritive protein hydrolysates from protein substrates such as whey, casein, soy, keratinous materials, and scraps from meat processing, as well as from fish processing discards; for the development of flavor during ripening of dairy products; and for the production of animal fodder from keratinous waste materials generated from meat and fish processing (Dalev 1994, Wilkinson and Kilcawley 2005). Other industrial applications of serine proteases include their use for the regioselective esterification of sugars, resolution of racemic mixtures of amino acids, and production of synthetic peptides to serve as pharmaceutical drugs and vaccines (Chen et al. 1999, Guzm an et al. 2007, Barros et al. 2009). Serine proteases are also used commercially for the treatment and bating of leather to remove undesirable pigments and hair from leather (Valera et al. 1997, Adg

Sulfhydryl Proteases
The sulfhydryl (also known as thiol or cysteine) proteases are so called because they contain intact sulfhydryl groups in their active sites and are inhibited by thiol reagents such as alkylating agents and heavy metal ions. Examples include plant types like actinidain, papain, bromelain, chymopapain, and ficin; animal types like cathepsins B and C, calpains, and caspases; and clostripain, protease 1 and others from various microorganisms such asStreptococcussp (Whitaker 1994), thermophilicBacillus sp (Almeida do Nascimento and Martins 2004), andPyrococcussp (Morikawa et al. 1994). The activities of this group of enzymes are enhanced by dithiothreitol and its epimer dithioerythritol, cysteine, and 2mercaptoethanol, and they are inhibited by compounds such as iodoacetate,pchloromercuribenzoic acid, cystatin, leupeptin, andN-ethylmaleimide. The enzymes in this group tend to be optimally active around neutral pH (i.e., pH 67.5) and are relatively heat stable, which accounts for their use in meat tenderizers. Thiol proteases act to degrade myofibril protein and collagen fibers to make meats tender. Meat tenderness is a very important factor in meat quality evaluation. Ficin tends to have the greatest effect on myofibril protein hydrolysis due to its relatively higher thermal stability and broader specificity, followed by bromelain and papain in that order, although bromelain tends to degrade collagen fibers more extensively (Miyada and Tappel 1956). In commercial application, papain and bromelain are more frequently used because the higher hydrolytic capacity of ficin often results in excessive softening and mushiness in the treated meat

Metalloproteases The term metalloproteases is used to encompass those proteases that have metal ions in their active sites. These enzymes typically require an essential metal ion (e.g., Ni 2+ ,Mg 2+ ,Mn 2+ ,Ca 2+ , Zn 2+ , and Co 2+ ) for functional activity. Urease is an example of the nickel (Ni 2+ )-containing metalloenzymes. It catalyzes the breakdown of urea into carbon dioxide (CO2) and ammonia (NH3). The urease-catalyzed reaction occurs as follows: (NH2) 2CO+H2OCO2+2NH3 The NH3 thus produced functions to neutralize gastric acid in the stomach. Urease is found in bacteria, yeast, and several higher plants. The major sources of urease include the Jack bean (Canavalia ensiformis) plant and the bacteria B. pasteurii. The enzyme is also found in large quantities in soybeans and other plant seeds, in certain animal tissues (e.g., liver, blood, and muscle), in intestinal bacteria (e.g., Eubacterium

N-Acetylaspartylglutamate+H2O Glutamic acid+N-Acetylaspartate while glutamyl aminopeptidase (also known as aminopeptidase A) catalyzes the hydrolysis of glutamic and aspartic acid residues from the N-termini of polypeptides, e.g., Aspartic acid-polypeptide+H2O Aspartic acid+Polypeptide Glutamyl

aminopeptidase is important because of its capacity to degrade angiotensin II into angiotensin III, which promote vasoconstriction and high blood pressure. Both

glutamate carboxypeptidase and glutamyl aminopeptidase are present in membranes.

Another important Zn 2+ -containing metalloprotease is the insulin-degrading enzyme that is found in humans and degrades short chain polypeptides such as the-chain of insulin, glucagon, transforming growth factor, and endorphins. Collagenases are also Zn 2+ dependent and they act to break down the peptide bonds in collagen. Cobalt (Co 2+ )-containing metalloproteases include methylmalonyl coenzyme A mutase that converts methyl malonyl-CoA to succinyl-CoA, which is involved in the extraction of energy from

proteins and fats; other Co 2+ -containing metalloenzymes are methionine

aminopeptidase and nitrile hydratase (Kobayashi and Shimizu 1999). Sources of metalloproteases include those derived from animals, like carboxypeptidases A and B,